(131 days)
No
The description details a standard real-time PCR assay and its associated instrument and software, with no mention of AI or ML for data analysis or interpretation. The analysis of results is based on Ct and Tm values, which are standard PCR metrics.
No
This device is an in vitro diagnostic test designed to detect and identify specific bacterial nucleic acids, aiding in diagnosis, rather than directly treating or preventing disease.
Yes
The "Intended Use / Indications for Use" section explicitly states that the ARIES® Bordetella Assay is a "qualitative in vitro diagnostic test for the direct detection and identification of Bordetella pertussis (B. pertussis) and Bordetella parapertussis (B. parapertussis) nucleic acid" which "aids in the diagnosis of B. pertussis and B. parapertussis respiratory infection". The "Device Description" also refers to it as a "qualitative in vitro diagnostic test system."
No
The device description explicitly states that the system includes an assay-specific cassette, which is a disposable, single-use cassette containing physical reagents for nucleic acid purification and PCR. This indicates the device is not solely software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The "Intended Use / Indications for Use" section explicitly states that the ARIES® Bordetella Assay is a "qualitative in vitro diagnostic test".
- Device Description: The "Device Description" also refers to it as a "qualitative in vitro diagnostic test system".
- Function: The device is designed to detect and identify specific nucleic acids (DNA) from human specimens (nasopharyngeal swabs) to aid in the diagnosis of a disease (respiratory infection attributable to B. pertussis or B. parapertussis). This is the core function of an in vitro diagnostic device.
- Specimen Type: It uses human biological specimens (nasopharyngeal swabs).
- Testing Location: It is intended for use in laboratory settings (hospital, reference or state laboratory settings), not for point-of-care use, which is typical for many IVDs.
The provided text clearly and repeatedly identifies the device as an in vitro diagnostic test.
N/A
Intended Use / Indications for Use
The ARIES® Bordetella Assay is a real-time polymerase chain reaction (PCR) based qualitative in vitro diagnostic test for the direct detection and identification of Bordetella pertussis (B. pertussis) and Bordetella parapertussis (B. parapertussis) nucleic acid in nasopharyngeal swab (NPS) specimens obtained from individuals suspected of having a respiratory tract infection attributable to B. pertussis or B. parapertussis.
The ARIES® Bordetella Assay targets the B. pertussis toxin promoter and the B. parapertussis IS1001 insertion element in the genomes. When clinical factors suggest that B. pertussis or B. parapertussis may not be the cause of respiratory infection, other clinically appropriate investigation(s) should be carried out in accordance with published guidelines.
Negative results for the ARIES® Bordetella Assay do not preclude B. pertussis or B. parapertussis infection and positive results do not co-infections with other respiratory pathogens. The direct detection and identification of B. pertussis and B. parapertussis nucleic acids from symptomatic patients aids in the diagnosis of B. pertussis and B. parapertussis respiratory infection in conjunction with other clinical findings and epidemiological information.
The ARIES® Bordetella Assay is indicated for use with the ARIES® Systems.
Product codes
OZZ
Device Description
The ARIES® Bordetella Assay is a polymerase chain reaction (PCR)-based qualitative in vitro diagnostic test system that consists of the ARIES® System or the ARIES® M1 System with their included ARIES® Software, an assay-specific cassette, and an assay-specific protocol file. The ARIES® Bordetella Assay cassette is a disposable, single-use cassette containing nucleic acid purification reagents, internal sample process control (SPC), and an assay-specific master mix capable of performing the designated assay on one sample. The ARIES® Bordetella Assay cassette directly detects and identifies B. pertussis and B. parapertussis DNA from nasopharyngeal swab (NPS) specimens collected from the human nasopharynx region.
Nasopharyngeal swab specimens are collected from patients using a commercially available E-Swab™ (Nylon® Flocked Swab along with modified Liquid Amies) or a commercially available nasopharyngeal swab (i.e. rayon, flocked, nylon, plastic shaft, etc.) placed into an approved transport media (i.e UTM, M5, M6, or equivalent). The specimen is then transported to the laboratory for testing. The specimen is lysed and nucleic acid is extracted using an ARIES® System. An extractable sample processing control (SPC) target is present in the ARIES® Bordetella Assay cassette and is processed with the specimen. The SPC controls for specimen lysis, for recovery of extracted nucleic acid, for inhibitory substances and for PCR reagent and instrument integrity. The Ct value of the SPC is designed to verify proper specimen lysis and nucleic acid extraction, to identify PCR inhibition, if any, and verify proper function of the extraction system and real-time instrument. The Tm value of the SPC is used as a reference for determining the target Tm.
The extracted nucleic acid and SPC are transferred via magnetic beads through the cassette to the ARIES® Bordetella Assay lyophilized PCR reagents in the PCR tube that contains primer pairs specific to the B. pertussis toxin promoter (ptxA-pr), the B. parapertussis IS1001 insertion element, and the SPC sequence. Each of the primer pairs are labeled with a distinct fluorophore and detected in distinct channels of the ARIES® Systems. PCR amplification is performed and assay fluorescence is monitored. Incorporation of a quencher-labeled nucleotide results in a decrease in fluorescence for the associated primer pair. Following amplification, the reaction is slowly heated to separate the fluorescent-labeled strand from the quencher-labeled strand, a process that results in an increase in the fluorescence signal. The reaction fluorescence is measured during this process and the temperature at which the change in fluorescence is the maximum Tm of the amplicon. The strands of the amplicons will separate at a specific melting temperature (Tm) and an increase in fluorescence is observed. The instrument fluorescence output is analyzed and test results are determined using the ARIES® System software and the ARIES® Bordetella Assay protocol and run files. ARIES® Bordetella Assay results may be reported from the ARIES® Software or from the optional SYNCT® Software.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Nasopharyngeal swab (NPS) specimens obtained from the human nasopharynx region.
Indicated Patient Age Range
pediatric and adult patients
Intended User / Care Setting
Not Found
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
Prospective Study:
- Sample Size: A total of 1052 unique specimens.
- Data Source: Leftover de-identified nasopharyngeal swab (NPS) specimens prospectively collected from pediatric and adult patients suspected of having respiratory tract infection attributable to B. pertussis or B. parapertussis. Collection occurred from July through November 2016 from five geographically distinct clinical sites within the United States.
- Annotation Protocol: The performance of ARIES® Bordetella Assay was compared to a composite comparator assay consisting of two well-characterized real-time PCR assays (for each bacterial pathogen) followed by confirmation of positive PCR amplification product with bi-directional sequencing. Comparator PCR assays for B. pertussis and B. parapertussis targeted unique sequences within the promoter region of the ptxA gene and IS1001 insertion region (respectively) that were different than those targeted by the ARIES® Bordetella Assay. Specimens were characterized as positive for B. pertussis or B. parapertussis if one out of two comparator PCR assays was positive (Ct values ≤40) and confirmed by bi-directional sequencing, or if both comparator PCR assays were positive. Specimens were characterized as B. pertussis or B. parapertussis negative if one out of two comparator PCR assays was negative (Ct values >40) and confirmed by bi-directional sequencing, or if both comparator PCR assays were negative. Comparator real-time PCR and bi-directional sequencing assays were performed at a centralized testing facility.
Supplemental Study (Banked and Contrived Specimens):
- Sample Size: B. pertussis (N=37) and B. parapertussis (N=20) positive banked (pre-selected) specimens, and contrived B. parapertussis specimens (N=50). Tested along with an equal number of unique negative clinical specimens.
- Data Source: Pre-selected B. pertussis specimens were collected at six (6) clinical sites in the United States while pre-selected B. parapertussis specimens were collected at three (3) sites also located in the United States. Contrived B. parapertussis samples were prepared by spiking well-characterized bacterial strains into individual negative clinical samples (NP swabs) at clinically relevant titers.
- Annotation Protocol: The presence of the expected bacterial target in each of the pre-selected and contrived specimens was confirmed by comparator real-time PCR and bi-directional sequencing assays. In addition, bacterial organism concentrations in contrived specimens were verified by culture methods (plating and colony count). To minimize bias, pre-selected and contrived specimens were tested along with an equal number of unique negative clinical specimens in a randomized, blinded fashion at three (3) external testing sites.
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Reproducibility/Precision/Repeatability:
- Study Type: Reproducibility study (site-to-site, lot-to-lot) and within-laboratory precision/repeatability.
- Sample Size:
- Site Reproducibility: 30 replicates per target type (B. pertussis low positive, moderate positive, B. parapertussis low positive, moderate positive, negative) at each of 3 sites (total 90 replicates per target type).
- Lot-to-Lot Reproducibility: 15 replicates per target type for each of 3 lots (total 45 replicates per target type).
- Within-Laboratory Precision/Repeatability: 30 replicates per target type (B. pertussis low positive, moderate positive, B. parapertussis low positive, moderate positive, negative).
- Key Results:
- Site Reproducibility: 100% positive for moderate positive samples, ≥ 95% positive for low positive samples, and 100% negative for negative samples across all sites.
- Lot-to-Lot Reproducibility: 100% positive for moderate positive samples, ≥ 95% positive for low positive samples, and 100% negative for negative samples across all lots.
- Within-Laboratory Precision/Repeatability: 100% positivity for all positive samples and 0% for negative samples, demonstrating repeatability across concentration levels.
Detection Limit (LoD):
- Study Type: Analytical sensitivity study.
- Sample Size: 20 replicates for each of 4 Bordetella strains (2 B. pertussis, 2 B. parapertussis).
- Key Results: LoD for each strain determined as the lowest concentration with ≥ 95% positivity. Overall assay LoD for B. pertussis is 1,800 CFU/mL and B. parapertussis is 213 CFU/mL.
Analytical Reactivity (Inclusivity):
- Study Type: Evaluation against different Bordetella strains.
- Sample Size: 18 Bordetella strains (11 B. pertussis, 7 B. parapertussis), tested in triplicate.
- Key Results: All 7 B. parapertussis strains detected with 100% positivity at 3x LoD. 9 of 11 B. pertussis strains detected with 100% positivity at 3x LoD. Two B. pertussis strains (ATCC 8478, ATCC 9797) were not detected due to identified nucleotide mismatches in primer binding regions, but an in-silico search suggested low prevalence and old collection dates for similar strains.
Analytical Specificity (Interfering Substances):
- Study Type: Evaluation of non-microbial substances.
- Sample Size: 3 replicates each of B. pertussis and B. parapertussis near LoD, and negative matrix, spiked with various substances.
- Key Results: None of the 17 substances tested showed interference.
Cross-Reactivity (Exclusivity):
- Study Type: Evaluation with common microorganisms.
- Sample Size: 65 unique microorganisms + multiple strains of Bordetella bronchiseptica and Bordetella holmesii (total 71 microorganisms), spiked into negative matrix and tested in triplicate.
- Key Results: 66 of 71 microorganisms yielded negative results. Five organisms (Fusobacterium necrophorum, Human Coronavirus OC43, Influenza B, Moraxella catarrhalis, Proteus vulgaris) generated a false positive result in 1 out of 6 replicates.
Microbial Interference (Co-Infection):
- Study Type: Evaluation of interfering organisms in presence of target.
- Sample Size: 65 unique microorganisms + multiple strains of Bordetella bronchiseptica and Bordetella holmesii (total 71 CROs) spiked into matrix containing B. pertussis or B. parapertussis near LoD concentrations, tested in triplicate. Also co-infection with B. pertussis and B. parapertussis evaluated (low/low, high/high, low/high, high/low concentrations).
- Key Results: B. pertussis was correctly detected in 3/3 replicates with 66 CROs (5 CROs required additional testing to achieve 5/6 detection). B. parapertussis was correctly detected in 3/3 replicates with all 71 CROs. All replicates in co-infection combinations yielded expected positivity.
Carry-Over/Cross-Contamination:
- Study Type: Evaluation by alternating high positive and negative samples.
- Sample Size: 30 high positive B. pertussis samples alternating with 30 negative samples across 5 consecutive runs.
- Key Results: No carry-over or cross contamination observed.
Clinical Performance:
- Study Type: Clinical trial (prospective study and supplemental study with banked/contrived specimens).
- Sample Size:
- Prospective Study: 1052 unique specimens.
- Supplemental Study: 37 B. pertussis positive, 20 B. parapertussis positive (pre-selected); 50 B. parapertussis (contrived).
- Standalone Performance: Not applicable (diagnostic test, comparison to composite reference method).
- Key Results:
- Prospective Study:
- B. pertussis: PPA = 93.8% (30/32), 95% CI: 79.2% - 99.2%. NPA = 98.9% (1009/1020), 95% CI: 98.1% - 99.5%.
- B. parapertussis: PPA = 100% (2/2), 95% CI: 15.8% - 100%. NPA = 99.8% (1048/1050), 95% CI: 99.3% - 100%.
- Supplemental Study (Pre-selected/Contrived):
- B. pertussis: PPA = 100% (37/37), 95% CI: 90.5% - 100%.
- B. parapertussis: PPA = 100% (20/20), 95% CI: 83.2% - 100%. (Note: 1 pre-selected specimen generated a false positive for B. parapertussis).
- Contrived B. parapertussis: PPA = 100% (50/50), 95% CI: 92.9% - 100%.
- Overall Combined Performance:
- B. pertussis: PPA = 97.1 (67/69), NPA = 99.0% (1086/1097).
- B. parapertussis: PPA = 100% (72/72), NPA = 99.7% (1191/1194).
- Prospective Study:
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Clinical Performance - Prospective Study:
- B. pertussis:
- Positive Percent Agreement (PPA): 93.8% (30/32) with a lower bound of the 95% confidence interval of 79.2%.
- Negative Percent Agreement (NPA): 98.9% (1009/1020) with a lower bound 95% confidence interval of 98.1%.
- B. parapertussis:
- Positive Percent Agreement (PPA): 100% (2/2) with a lower bound of the 95% confidence interval of 15.8%.
- Negative Percent Agreement (NPA): 99.8% (1048/1050) with a lower bound 95% confidence interval of 99.3%.
Clinical Performance - Pre-selected/Contrived Specimens:
- B. pertussis:
- Positive Percent Agreement (PPA): 100% (37/37); 95% confidence interval: 90.5% - 100%.
- B. parapertussis:
- Positive Percent Agreement (PPA): 100% (20/20); 95% confidence interval: 83.2% - 100%.
- Contrived B. parapertussis Positive Percent Agreement (PPA): 100% (50/50); 95% confidence interval 92.9%- 100%.
Overall Combined Performance (Prospective, Pre-selected, Contrived Specimens):
- B. pertussis:
- PPA: 97.1% (67/69)
- NPA: 99.0% (1086/1097)
- B. parapertussis:
- PPA: 100% (72/72)
- NPA: 99.7% (1191/1194)
Predicate Device(s)
Reference Device(s)
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information
Not Found
§ 866.3980 Respiratory viral panel multiplex nucleic acid assay.
(a)
Identification. A respiratory viral panel multiplex nucleic acid assay is a qualitative in vitro diagnostic device intended to simultaneously detect and identify multiple viral nucleic acids extracted from human respiratory specimens or viral culture. The detection and identification of a specific viral nucleic acid from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory viral infection when used in conjunction with other clinical and laboratory findings. The device is intended for detection and identification of a combination of the following viruses:(1) Influenza A and Influenza B;
(2) Influenza A subtype H1 and Influenza A subtype H3;
(3) Respiratory Syncytial Virus subtype A and Respiratory Syncytial Virus subtype B;
(4) Parainfluenza 1, Parainfluenza 2, and Parainfluenza 3 virus;
(5) Human Metapneumovirus;
(6) Rhinovirus; and
(7) Adenovirus.
(b)
Classification. Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay;”
(2) For a device that detects and identifies Human Metapneumovirus, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Human Metapneumovirus (hMPV) Using Nucleic Acid Assays;” and
(3) For a device that detects and differentiates Influenza A subtype H1 and subtype H3, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Detection and Differentiation of Influenza A Virus Subtypes Using Multiplex Nucleic Acid Assays.” See § 866.1(e) for the availability of these guidance documents.
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Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized graphic of three human profiles facing right, stacked on top of each other. The profiles are rendered in black and have a flowing, abstract design. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular fashion around the graphic.
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
May 2, 2017
Luminex Corporation Kate Linak Regulatory Affairs Scientist 12212 Technology Blvd Austin, Texas 78727
Re: K163626
Trade/Device Name: ARIES Bordetella Assay Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory Viral Panel Multiplex Nucleic Acid Assay Regulatory Class: Class II Product Code: OZZ Dated: April 4, 2017 Received: April 4, 2017
Dear Ms. Linak:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA), You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices. good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the
1
electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
If you desire specific advice for your device on our labeling regulation (21 CFR Part 801 and Part 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to
http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely,
Kristian M. Roth -S
For : Uwe Scherf, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
2
Indications for Use
510(k) Number (if known) K163626
Device Name ARIES® Bordetella Assay
Indications for Use (Describe)
The ARIES® Bordetella Assay is a real-time polymerase chain reaction (PCR) based qualitative in virro diagnostic test for the direct detection and identification of Bordetella pertussis) and Bordetella parapertussis (B. parapertussis) nucleic acid in nasopharyngeal swab (NPS) specimens obtained from individuals suspected of having a respiratory tract infection attributable to B. pertussis or B. parapertussis.
The ARIES® Bordetella Assay targets the B. pertussis toxin promoter and the B. parapertussis IS1001 insertion element in the genomes. When clinical factors suggest that B. pertussis or B. parapertussis may not be the cause of respiratory infection, other clinically appropriate investigation(s) should be carried out in accordance with published guidelines.
Negative results for the ARIES® Bordetella Assay do not preclude B. pertussis infection and positive results do not rule out co-infections with other respiratory pathogens. The direction and identification of B. pertussis and B. parapertussis nucleic acids from symptomatic patients aids in the diagnosis of B. perfussis and B. parapertussis respiratory infection with other clinical findings and epidemiological information.
The ARIES® Bordetella Assay is indicated for use with the ARIES® Systems.
| Type of Use (Select one or both, as applicable) | |
|---|---|
| [2] Prescription Use (Part 21 CFR 801 Subpart D) | O Over-The-Counter Use (21 CFR 801 Subpart C) |
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Executive Summary 510(k)
This Executive Summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.
A. 510(k) Number:
B. Purpose for Submission:
Clearance of the ARIES® Bordetella Assay for use with the ARIES® Systems.
C. Measurand:
Bordetella pertussis toxin promoter, Bordetella parapertussis IS1001 insertion element in respective genomes.
D. Type of Test:
Qualitative Real Time Polymerase Chain Reaction (PCR).
E. Applicant:
Luminex Corporation
F. Proprietary and Established Names:
ARIES® Bordetella Assay
G. Regulatory Information:
| Product
| Code | Classification | Regulation Section | Panel |
|---|---|---|---|
| OZZ | Class II | 21 CFR 866.3980—Respiratory viral panel | |
| multiplex nucleic acid assay | Microbiology | ||
| (83) |
H. Intended Use:
1. Intended use(s):
The ARIES® Bordetella Assay is a real-time polymerase chain reaction (PCR) based qualitative in vitro diagnostic test for the direct detection and identification of Bordetella pertussis (B. pertussis) and Bordetella parapertussis (B. parapertussis) nucleic acid in
4
nasopharyngeal swab (NPS) specimens obtained from individuals suspected of having a respiratory tract infection attributable to B. pertussis or B. parapertussis.
The ARIES® Bordetella Assay targets the B. pertussis toxin promoter and the B. parapertussis IS1001 insertion element in the genomes. When clinical factors suggest that B. pertussis or B. parapertussis may not be the cause of respiratory infection, other clinically appropriate investigation(s) should be carried out in accordance with published guidelines.
Negative results for the ARIES® Bordetella Assay do not preclude B. pertussis or B. parapertussis infection and positive results do not co-infections with other respiratory pathogens. The direct detection and identification of B. pertussis and B. parapertussis nucleic acids from symptomatic patients aids in the diagnosis of B. pertussis and B. parapertussis respiratory infection in conjunction with other clinical findings and epidemiological information.
The ARIES® Bordetella Assay is indicated for use with the ARIES® Systems.
-
- Indication(s) for use:
Same as intended use.
- Indication(s) for use:
-
- Special conditions for use statement(s):
For prescription use only.
- Special conditions for use statement(s):
-
- Special instrument requirements:
For use with the ARIES® Systems.
- Special instrument requirements:
l. Device Description:
The ARIES® Bordetella Assay is a polymerase chain reaction (PCR)-based qualitative in vitro diagnostic test system that consists of the ARIES® System or the ARIES® M1 System with their included ARIES® Software, an assay-specific cassette, and an assay-specific protocol file. The ARIES® Bordetella Assay cassette is a disposable, single-use cassette containing nucleic acid purification reagents, internal sample process control (SPC), and an assay-specific master mix capable of performing the designated assay on one sample. The ARIES® Bordetella Assay cassette directly detects and identifies B. pertussis and B. parapertussis DNA from nasopharyngeal swab (NPS) specimens collected from the human nasopharynx region.
Nasopharyngeal swab specimens are collected from patients using a commercially available E-Swab™ (Nylon® Flocked Swab along with modified Liquid Amies) or a commercially available nasopharyngeal swab (i.e. rayon, flocked, nylon, plastic shaft, etc.) placed into an approved transport media (i.e UTM, M5, M6, or equivalent). The specimen is then transported to the laboratory for testing. The specimen is lysed and nucleic acid is extracted using an ARIES® Executive Summary Page 2 of 20
5
System. An extractable sample processing control (SPC) target is present in the ARIES® Bordetella Assay cassette and is processed with the specimen. The SPC controls for specimen lysis, for recovery of extracted nucleic acid, for inhibitory substances and for PCR reagent and instrument integrity. The Ct value of the SPC is designed to verify proper specimen lysis and nucleic acid extraction, to identify PCR inhibition, if any, and verify proper function of the extraction system and real-time instrument. The Tη value of the SPC is used as a reference for determining the target Tm.
The extracted nucleic acid and SPC are transferred via magnetic beads through the cassette to the ARIES® Bordetella Assay lyophilized PCR reagents in the PCR tube that contains primer pairs specific to the B. pertussis toxin promoter (ptxA-pr), the B. parapertussis IS1001 insertion element, and the SPC sequence. Each of the primer pairs are labeled with a distinct fluorophore and detected in distinct channels of the ARIES® Systems. PCR amplification is performed and assay fluorescence is monitored. Incorporation of a quencher-labeled nucleotide results in a decrease in fluorescence for the associated primer pair. Following amplification, the reaction is slowly heated to separate the fluorescent-labeled strand from the quencher-labeled strand, a process that results in an increase in the fluorescence signal. The reaction fluorescence is measured during this process and the temperature at which the change in fluorescence is the maximum Tm of the amplicon. The strands of the amplicons will separate at a specific melting temperature (Tm) and an increase in fluorescence is observed. The instrument fluorescence output is analyzed and test results are determined using the ARIES® System software and the ARIES® Bordetella Assay protocol and run files. ARIES® Bordetella Assay results may be reported from the ARIES® Software or from the optional SYNCT® Software.
J. Substantial Equivalence Information:
1. Predicate device name(s):
illumigene® Pertussis DNA Amplification Assay (manufactured by Meridian Bioscience, Inc.)
2. Predicate 510(k) number(s):
3. Comparison with predicate:
The following table compares the ARIES® Bordetella Assay to Meridian Bioscience, Inc.'s illumiqene® Pertussis DNA Amplification Assay (K133673). Table 11.1 shows similarities between the new device and the predicate, while Table 11.2 shows the differences.
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| Similarities | ||
|---|---|---|
| Attribute | New Device | Predicate Device (K133673) |
| Intended Use | The ARIES® Bordetella Assay is a real-time polymerase chain reaction (PCR) based qualitative in vitro diagnostic test for the direct detection and identification of Bordetella pertussis ( B. pertussis ) and Bordetella parapertussis ( B. parapertussis ) nucleic acid in nasopharyngeal swab (NPS) specimens obtained from individuals suspected of having a respiratory tract infection attributable to B. pertussis or B. parapertussis . |
The ARIES® Bordetella Assay targets the B. pertussis toxin promoter and the B. parapertussis IS1001 insertion element in the genomes. When clinical factors suggest that B. pertussis or B. parapertussis may not be the cause of respiratory infection, other clinically appropriate investigation(s) should be carried out in accordance with published guidelines.
Negative results for the ARIES® Bordetella Assay do not preclude B. pertussis or B. parapertussis infection and positive results do not rule out co-infections with other respiratory pathogens. The direct detection and identification of B. pertussis and B. parapertussis nucleic acids from symptomatic patients aids in the diagnosis of B. pertussis and B. parapertussis respiratory infection in conjunction with other clinical findings and epidemiological information.
The ARIES® Bordetella Assay is indicated for use with the ARIES® Systems. | The illumigene® Pertussis DNA Amplification Assay, performed on the illumipro-10™, is a qualitative in vitro diagnostic test for the direct detection of Bordetella pertussis in human nasopharyngeal swab samples taken from patients suspected of having respiratory tract infection attributable to Bordetella pertussis.
The illumigene Pertussis assay utilizes loop-mediated isothermal DNA amplification (LAMP) technology to detect Bordetella pertussis by targeting the IS481 insertional element of the Bordetella pertussis genome. The IS481 insertional element can also be found in Bordetella holmesii and Bordetella bronchiseptica strains. Respiratory infection with Bordetella pertussis, Bordetella holmesii or Bordetella bronchiseptica may yield positive test results in IS481 assays. B. holmesii infection may cause clinical illness similar to B. pertussis , and mixed outbreaks involving both B. pertussis and B. holmesii infection have been reported. Additional testing should be performed if necessary to differentiate B. holmesii and B. pertussis . B. bronchiseptica is a rare cause of infection in humans. When clinical factors suggest that B. pertussis may not be the cause of respiratory infection, other clinically appropriate investigation(s) should be carried out in accordance with published guidelines.
Negative results for the illumigene® Pertussis DNA Amplification Assay do not preclude Bordetella pertussis infection and positive results do not rule out co-infection with other respiratory pathogens. Results from the illumigene Pertussis assay should be used in conjunction with information obtained during the patient's clinical evaluation as an aid in diagnosis of Bordetella pertussis |
| Similarities | | |
| Attribute | New Device | Predicate Device (K133673) |
| | | infection and should not be used as the
sole basis for treatment or other patient
management decisions.
illumigene Pertussis is intended for use
in hospital, reference or state laboratory
settings. The device is not intended for
point-of-care use. |
| Sample type | Nasopharyngeal swabs (NPS) | Nasopharyngeal swabs (NPS) |
| Assay results | Qualitative | Qualitative |
| Analyte | DNA | DNA |
Table 11.1: Similarities between New Device and Predicate
7
Table 11.2: Differences between New Device and Predicate
| Differences | ||
|---|---|---|
| Attribute | New Device | Predicate Device (K120413) |
| Extraction Method | Automated by the ARIES® Systems | Manual extraction |
| Organisms Detected | B. pertussis and B. parapertussis | Bordetella pertussis |
| B. pertussis Target | toxin promoter (ptxA-pr) | IS481 Insertional Element |
| B. parapertussis Target | IS1001 insertion element | N/A |
| Detection | Different fluorescent reporter dyes for each target and melt analysis. | |
| Fluorescence Emissions and Detection | Measurement of magnesium pyrophosphate forming a precipitate in the reaction mixture. | |
| Visible Light Transmission | ||
| Assay format | Real-time PCR | DNA Amplification; Loop-Mediated Isothermal Amplification (LAMP) |
| Controls | Internal Control: Sample processing control (SPC) | Internal Control Provided; External positive control included in illumigene Pertussis External Control Kit |
| Instrument | ARIES® System, ARIES® M1 System | illumipro-10 |
K. Standards/Guidance Documents Referenced:
Not applicable.
L. Test Principle:
The ARIES® Bordetella Assay chemistry is based on an expanded genetic alphabet technology, consisting of synthetic DNA base pair 2'-deoxy-5-methyl-isocytidine (iC): 2'-deoxyisoguanosine (iG). The isobases (iC and iG) pair specifically with each other and not with natural nucleotides. In addition, isobases are efficiently incorporated during PCR amplification, a quenchermodified iGTP is incorporated by the polymerase opposite an iC and a fluorophore reporter attached to a PCR primer. If the target is present and is amplified, assay fluorescence decreases with every cycle as amplification product accumulates. The decrease in assay fluorescence is
8
monitored in real time using the ARIES® Systems. Following PCR, the amplification products are thermally denatured and assay fluorescence is monitored. The strands of the amplification products are separated and assay fluorescence increases, thus enabling determination of the melting temperature (Tm) of the amplicon.
M. Performance Characteristics:
1. Analytical performance:
Reproducibility/Precision/Repeatability: a.
Reproducibility of the ARIES® Bordetella Assay was evaluated by testing one lot of ARIES® Bordetella Assay Cassettes on two ARIES® Systems by two operators at each of three sites, 2 external clinical and 1 internal, on five non-consecutive days. A blinded and randomized reproducibility panel was prepared and sent to these sites by an independent operator that consisted of B. pertussis low positive, B. pertussis moderate positive, B. parapertussis low positive, B. parapertussis moderate positive and B. pertussis and B. parapertussis negative sample diluted into the negative natural nasopharyngeal matrix. Each panel member was tested in triplicate by each operator each day of testing. The reproducibility study are shown in Tables 11.3 and 11.4. The results showed that the reproducibility of the ARIES® Bordetella Assay across the three sites met the acceptance criteria of 100% positive for moderate positive samples, ≥ 95% positive for low positive samples, and 100% negative for negative samples.
| Targets | Site 1 | Site 2 | Site 3 | |||
|---|---|---|---|---|---|---|
| Positivity | Positivity | Positivity | ||||
| B. pertussis (Low Positive) | 30/30 | 100% | 30/30 | 100% | 30/30 | 100% |
| B. pertussis (Moderate Positive) | 30/30 | 100% | 30/30 | 100% | 30/30 | 100% |
| B. parapertussis (Low Positive) | 30/30 | 100% | 30/30 | 100% | 30/30 | 100% |
| B. parapertussis (Moderate Positive) | 30/30 | 100% | 30/30 | 100% | 30/30 | 100% |
| Negative | 0/30 | 0% | 0/30 | 0% | 0/30 | 0% |
Table 11.3: ARIES® Bordetella Assay Site Reproducibility Results
Table 11.4: Reproducibility Panel Overall Results
| Targets | Positivity | 95% Confidence Interval | ||
|---|---|---|---|---|
| Lower Limit | Upper Limit | |||
| B. pertussis (Low Positive) | 90/90 | 100% | 96.0% | 100.0% |
| B. pertussis (Moderate Positive) | 90/90 | 100% | 96.0% | 100.0% |
| B. parapertussis (Low Positive) | 90/90 | 100% | 96.0% | 100.0% |
| B. parapertussis (Moderate Positive) | 90/90 | 100% | 96.0% | 100.0% |
| Negative | 0/90 | 0% | 0.0% | 4.0% |
9
Lot-to-Lot Reproducibility of the ARIES® Bordetella Assay was evaluated by one operator using one ARIES® instrument to test three lots of the ARIES® Bordetella Assay Cassettes. A reproducibility panel consisting of B. pertussis low positive, B. pertussis moderate positive, B. parapertussis low positive, B. parapertussis moderate positive, and B. pertussis and B. parapertussis negative sample diluted into the negative natural nasopharyngeal matrix were prepared and tested to evaluate the lot-to-lot reproducibility. Three replicates of each sample concentration were run five times (for a total of fifteen replicates) for each lot of the ARIES® Bordetella Assay Cassette. The identity of these samples was blinded to the operator. The results of the study are shown in Table 11.5. The test results showed that the reproducibility across the three ARIES® Bordetella Assay Cassette lots met the acceptance criteria of 100% positive for moderate positive samples, ≥ 95% positive for low positive samples, and 100% negative for negative samples.
| Positivity | ||||
|---|---|---|---|---|
| Target Type | Lot AA1445 | Lot AA1485 | Lot AA1505 | Overall |
| B. pertussis (Low Positive) | 100% (15/15) | 100% (15/15) | 100% (15/15) | 100% (45/45) |
| B. pertussis (Moderate Positive) | 100% (15/15) | 100% (15/15) | 100% (15/15) | 100% (45/45) |
| B. parapertussis (Low Positive) | 100% (15/15) | 100% (15/15) | 100% (15/15) | 100% (45/45) |
| B. parapertussis (Moderate Positive) | 100% (15/15) | 100% (15/15) | 100% (15/15) | 100% (45/45) |
| Negative | 0% (0/15) | 0% (0/15) | 0% (0/15) | 0% (0/45) |
Table 11.5: ARIES® Bordetella Assay Lot-to-Lot Reproducibility Determination Results
Within-laboratory precision/repeatability for the ARIES® Bordetella Assay was evaluated by testing B. pertussis and B. parapertussis samples at various concentration levels across multiple days utilizing multiple operators, multiple ARIES® Systems and one lot of the ARIES® Bordetella Assay Cassette. Study samples were prepared that consisted of B. pertussis and B. parapertussis culture diluted into negative natural nasopharyngeal matrix at five concentration levels - moderate positive B. pertussis, low positive B. pertussis, moderate positive B. parapertussis, low positive B. parapertussis, and a B. pertussis and B. parapertussis negative samples were blinded to the operator and tested in triplicate by two operators across five non-consecutive days. The results of the study shown in Table 11.6 demonstrate that results obtained with the ARIES® Bordetella Assay between multiple operators using multiple instruments within a laboratory run across multiple days are repeatable across a range of Bordetella concentration levels.
Table 11.6: ARIES® Bordetella Assay Within Laboratory Precision/ Repeatability Results
| Target Type | Expected Positivity | Positivity | 95% Confidence Interval |
|---|---|---|---|
| B. pertussis (Low Positive) | Approximately 95% | 100% (30/30) | 88.4% - 100.0% |
| B. pertussis (Moderate Positive) | 100% | 100% (30/30) | 88.4% - 100.0% |
| B. parapertussis (Low Positive) | Approximately 95% | 100% (30/30) | 88.4% - 100.0% |
| B. parapertussis (Moderate Positive) | 100% | 100% (30/30) | 88.4% - 100.0% |
| Negative | 0% | 0% (0/30) | 0.0% - 11.6% |
10
-
b. Linearity/assay reportable range:
Not applicable. The ARIES® Bordetella Assay is a qualitative assay. -
ﻥ Traceability, Stability, Expected values (controls, calibrators, or methods):
Stability:
Specimen Stability (Fresh vs. Frozen)
The performance equivalency of the ARIES® Bordetella Assay was assessed using contrived Bordetella specimens when tested from the fresh state (i.e. unfrozen) and specimens that were tested after being stored at -65 to -95°C (frozen). Equivalency was evaluated using samples consisting of Bordetella pertussis and Bordetella parapertussis culture diluted independently into natural negative human nasopharyngeal swab matrix at three test concentrations. A total of 120 samples were tested for each Bordetella strain of which half were tested immediately upon preparation (fresh) and the remaining samples were frozen at -65 to -95°C and tested 24 to 48 hours later. Six replicates of natural negative human nasopharyngeal swab matrix were also tested fresh and frozen as a negative control. Of each set of fresh and frozen samples, 50% were targeted to 3x LoD, 25% were targeted to 10X LoD and the remaining 25% of the samples were targeted to 100X LoD.
The results of the study are shown in Table 11.7. Based on this study, no significant difference in the performance of ARIES® Bordetella Assay was observed between specimens tested fresh and specimens that were tested after being stored frozen at -65 to -95°C.
| Target type | Concentrations | Agreement with Expected Results | |
|---|---|---|---|
| Fresh Samples | Frozen Samples | ||
| B. pertussis | 3x LoD | 100% (30/30) | 100% (30/30) |
| B. pertussis | 10x LoD | 100% (15/15) | 100% (15/15) |
| B. pertussis | 100x LoD | 100% (15/15) | 100% (15/15) |
| B. parapertussis | 3x LoD | 100% (30/30) | 100% (30/30) |
| B. parapertussis | 10x LoD | 100% (15/15) | 100% (15/15) |
| B. parapertussis | 100x LoD | 100% (15/15) | 100% (15/15) |
| Natural Negative Matrix | Negative | 100% (6/6) | 100% (6/6) |
Table 11.7: ARIES® Bordetella Assay Fresh vs. Frozen Contrived Specimen Stability Results
Shelf-Life Stability
A real time stability study was performed to evaluate the shelf life of ARIES® Bordetella Assay Cassettes. Stability was assessed by testing three replicates of Bordetella Extractable Control, 100x LoD Blend and three replicates of negative targets (Copan UTM) on three different lots of ARIES® Bordetella cassettes stored at two different temperatures (4°C (2 – 8
11
°C) and room temperature (15 – 30 °C)). The study was designed following guidelines listed in EP25-A, Evaluation of Stability of In Vitro Diagnostic Reagents; Approved Guideline and BS EN ISO 23640: 2013 - In vitro diagnostic medical devices - Evaluation of stability of in vitro diagnostic reagents. Acceptance criteria for stability at each time point and temperature were established as 100% positivity for all Bordetella replicates and 100% negativity for all negative replicates. Data for ARIES® Bordetella Assay Cassette stability has been collected up to 7 months and gave expected results indicating stability up to 7 months. Stability studies are on-going.
Controls:
Process Control
Each ARIES® Bordetella Assay cassette contains a Sample Process Control (SPC), which is processed with the sample and analyzed during the amplification reaction. The SPC verifies sample lysis, nucleic acid extraction, and proper reagent, cassette, ARIES® System, and assay protocol performance. The SPC has a known melting temperature (Tm) range and Ct range. Each time an assay is run, the system measures the temperature and fluorescence intensity of the SPC control to ensure the thermal and optical subsystems have remained in calibration.
External Controls
External controls should be tested according to guidelines or requirements of local, provincial and/or federal regulations or accreditation organizations. Reference Bordetella pertussis and Bordetella parapertussis strains or well characterized Bordetella pertussis and Bordetella parapertussis clinical isolates may be used as positive controls. The ARIES® Bordetella Assay Cassette Kit does not include external positive and negative controls.
d. Detection Limit:
A Limit of Detection (LoD) study was performed to evaluate the analytical sensitivity of the ARIES® Bordetella Assay using two strains each of B. pertussis and B. parapertussis diluted in natural negative nasopharyngeal (NP) swab matrix. Preliminary LoD concentrations were determined using serial dilutions of each Bordetella strain in natural negative nasopharyngeal (NP) swab matrix where each dilution was quantified using standard dilution and quantitative culture techniques. These preliminary LoD concentrations were confirmed by testing twenty (20) replicates of each strain. All Bordetella strain concentrations were verified by plating and colony counting (CFU/mL). The LoD for each Bordetella strain was determined as the lowest concentration that had a positivity rate of ≥ 95%. The final LoD concentrations for the four strains of Bordetella are shown in Table 11.8. The overall assay LoD for B. pertussis is 1,800 CFU/mL and B. parapertussis is 213 CFU/mL.
12
| Bordetella
Type | Strain | Concentration
(CFU/mL) | Positivity | 95% Confidence
Interval |
|-------------------------|---------|---------------------------|--------------|----------------------------|
| B. pertussis | A639 | 1,640 | 95% (19/20) | 75.1% - 99.9% |
| B. pertussis | BAA-589 | 1,800 | 95% (19/20) | 75.1% - 99.9% |
| B. parapertussis | A747 | 172 | 100% (20/20) | 83.2% - 100.0% |
| B. parapertussis | BAA-587 | 213 | 95% (19/20) | 75.1% - 99.9% |
Table 11.8: ARIES® Bordetella Assay Limit of Detection Results
e. Analytical Reactivity (Inclusivity)
The analytical reactivity (inclusivity) of the ARIES® Bordetella Assay was evaluated against eighteen (18) Bordetella strains; eleven B. pertussis and seven B. parapertussis strains. These strains differed from those that were tested as part of the Limit of Detection study. The specimens were prepared by diluting quantified cultured organism into pooled natural negative human nasopharyngeal swab matrix at a concentration of three times the confirmed limit of detection of the ARIES® Bordetella Assay and tested in triplicate. All seven strains of B. parapertussis were detected with 100% positivity at 3x LoD. Additionally, nine of the eleven strains of B. pertussis were detected with 100% positivity at 3x LoD while two strains, ATCC 8478 and ATCC 9797, were not detected at either 3x, 10x or 100x LoD. Sequencing of these two strains showed that both strains contained a similar nucleotide mismatch in both ARIES® B. pertussis primer binding regions, forward and reverse, which may impact the ability of the ARIES® Bordetella assay to detect these strains. An in-silico search of Bordetella pertussis sequences in the NCBI database identified a low prevalence of strains with similar mismatches (2.9%). Furthermore, all of these strains have decades old collection dates for strains with known human hosts-suggesting that these strains are no longer prevalent in the human population. Thus, there is a low risk of the ARIES® Bordetella Assay missing a circulating strain of B. pertussis. The results of the inclusivity testing are shown in Table 11.9.
| Inclusivity Strains | Test Concentrationb | Result | Positivity |
|---|---|---|---|
| ATCC BAA-1335 | 5,400 CFU/mL | B. pertussis Detected | 100% (3/3) |
| ATCC 8467 | 5,400 CFU/mL | B. pertussis Detected | 100% (3/3) |
| ATCC 12742 | 5,400 CFU/mL | B. pertussis Detected | 100% (3/3) |
| ATCC 12743 | 5,400 CFU/mL | B. pertussis Detected | 100% (3/3) |
| ATCC 51445 | 5,400 CFU/mL | B. pertussis Detected | 100% (3/3) |
| E431 | 5,400 CFU/mL | B. pertussis Detected | 100% (3/3) |
| ATCC 10380 | 5,400 CFU/mL | B. pertussis Detected | 100% (3/3) |
| NR-42457 | 5,400 CFU/mL | B. pertussis Detected | 100% (3/3) |
| NR-42460 | 5,400 CFU/mL | B. pertussis Detected | 100% (3/3) |
| ATCC 8478 | 180,000 CFU/mLa | B. pertussis Not Detected | 0% (3/3) |
| ATCC 9797 | 180,000 CFU/mLa | B. pertussis Not Detected | 0% (3/3) |
Table 11.9: ARIES® Bordetella Assay Analytical Reactivity (Inclusivity) Results
13
| Inclusivity Strains | Test Concentrationb | Result | Positivity |
|---|---|---|---|
| ATCC 9305 | 639 CFU/mL | B. parapertussis Detected | 100% (3/3) |
| ATCC 15237 | 639 CFU/mL | B. parapertussis Detected | 100% (3/3) |
| ATCC 15311 | 639 CFU/mL | B. parapertussis Detected | 100% (3/3) |
| ATCC 15989 | 639 CFU/mL | B. parapertussis Detected | 100% (3/3) |
| C510 | 639 CFU/mL | B. parapertussis Detected | 100% (3/3) |
| E595 | 639 CFU/mL | B. parapertussis Detected | 100% (3/3) |
| E838 | 639 CFU/mL | B. parapertussis Detected | 100% (3/3) |
4 Two strains of B. pertussis, ATCC 8478 and ATCC 9797 were not detected using the ARIES® Bordetella Assay up to 100x of the assay LoD.
6 B. pertussis and B. parapertussis strains were tested at 3x LoD.
f. Analytical specificity:
Interfering Substances:
The potential inhibitory effect of non-microbial substances expected to be found in human nasopharyngeal swab specimens was evaluated for the ARIES® Bordetella Assay. Three replicates each of Bordetella pertussis and Bordetella parapertussis were tested at concentrations near the assay LoD with a relative, high concentration of each non-microbial substance spiked into the contrived sample. Additionally, negative natural human nasopharyngeal swab matrix was spiked with the same concentration of each non-microbial substance and tested for assay interference. Bordetella pertussis and Bordetella parapertussis samples were contrived by independently diluting cultures into natural negative human nasopharyngeal swab matrix at a concentration of three times the limit of detection of the ARIES® Bordetella Assay for the strain tested.
The results of the study are shown in Table 11.10. Based on this study, none of the substances tested showed any interference with the ARIES® Bordetella Assay.
| Interfering Substance | Test Concentration |
|---|---|
| Benzocaine | 2.5% (w/v) |
| Budesonide | 25 mg/mL |
| Dexamethasone | 3 mg/mL |
| Flunisolide | 55 mg/mL |
| Fluticasone (Nasal Corticosteroids) | 5% (v/v) |
| FluMist | 10% (v/v) |
| Human Blood (EDTA) | 5% (v/v) |
| Menthol | 0.26% (w/v) |
| Mometasone | 2.5 mg/mL |
| Mucin protein | 1% (w/v) |
| Mupirocin | 2% (w/v) |
| Oseltamivir Phosphate (Anti-viral drugs) | 10 mg/mL |
| Oxymetazoline nasal spray (Afrin®) | 15% (v/v) |
| Phenylephrine | 0.3 mg/mL |
| Smokeless tobacco | 1% (w/v) |
Table 11.10: ARIES® Bordetella Assay Interfering Substance Results
14
| Interfering Substance | Test Concentration |
|---|---|
| Sodium chloride | 0.0065% (w/v) |
| Tobramycin (Antibacterial, systemic) | 0.6 mg/mL |
| Triamcinolone | 5.5 µg/mL |
| Zanamivir (Anti-viral drugs) | 5 mg/mL |
| Zicam Nasal gel (Histaminum hydrochoricum, Galphimia glauca, | |
| Luffa opperculata, Sulfur) | 5% (v/v) |
Cross-Reactivity (Exclusivity):
Cross reactivity for the ARIES® Bordetella Assay was assessed with 65 unique microorganisms that are common to the same human matrix as B. pertussis and or B. parapertussis infections, that cause infections that might present similar symptoms to B. pertussis and or B. parapertussis infections, and that could potentially interfere with the diagnostic capabilities of the ARIES® Bordetella Assay. Bacteria were tested at ≥10° CFU/mL or the highest available concentration, and viruses were tested at ≥10° TCID50/mL or the highest available concentration. These potential cross reactive organisms were spiked at high concentration into natural negative human nasopharyngeal swab matrix and tested in triplicate (n=3) on the ARIES® System. In addition to the 65 unique microorganisms, multiple strains of Bordetella bronchiseptica and Bordetella holmesii were also tested for crossreactivity for a total of 71 microorganisms. Of the 71 microorganisms tested, 66 yielded negative results for B. pertussis and B. parapertussis and thus are considered non-reactive with the ARIES® Bordetella Assay. Five organisms, Fusobacterium necrophorum, Human Coronavirus OC43, Influenza B, Moraxella catarrhalis and Proteus vulgaris generated a false positive result in 1 out of 6 replicates. All microorganisms evaluated are listed in Table 11.11.
Microbial Interference (Cross Reactivity)/Co-Infection:
Microbial interference for the ARIES® Bordetella Assay was assessed with 65 unique cross reactive microorganisms (CRO) that are commonly found in the same human matrix as B. pertussis and or B. parapertussis infections, that cause infections that might present similar symptoms to B. pertussis and or B. parapertussis infections, and could potentially interfere with the diagnostic capabilities of the ARIES® Bordetella Assay. Bacteria were tested at ≥10° CFU/mL or the highest available concentration, and viruses were tested at ≥107 TCIDso/mL or the highest available concentration. The potential interfering organisms were spiked into natural negative human nasopharyngeal swab matrix containing representative strains of Bordetella pertussis (BP) or Bordetella parapertussis (BPP) near the LoD concentration. All target strain + CRO samples were tested in triplicate (n=3) on the ARIES® System. In addition to the 65 unique microorganisms, multiple strains of Bordetella bronchiseptica and Bordetella holmesii were also tested for microbial interference. B. pertussis was correctly detected in 3/3 replicates when tested in the presence of 66 CROs, with 5 CROs, Bordetella bronchiseptica (strain 1 and strain 2), Bordetella petrii, Enterobacter aerogenes, Klebsiella pneumoniae requiring additional testing of 3 replicates per protocol. For these 5 CROs, B.
15
pertussis was detected in 5/6 replicates. B. parapertussis was correctly detected in 3/3 replicates when tested in the presence of all 71 CROs.
Microbial interference was also evaluated in a co-infection setting where near LoD concentration B. pertussis was tested with high concentration of B. parapertussis and viceversa. A low/low and a high/high combination of B. pertussis and B. parapertussis were also evaluated. All replicates in all combinations yielded expected positivity for B. pertussis and B. parapertussis. All microorganisms evaluated are listed in Table 11.11.
| Microorganism | |
|---|---|
| 1 | Acinetobacter baumanii |
| 2 | Acinetobacter calcoaceticus |
| 3 | Acinetobacter Iwoffii9 |
| 4 | Adenovirus 7A |
| 5 | Adenovirus Type 1 |
| 6 | Adenovirus Type 3 |
| 7 | Arcanobacterium haemolyticum9 |
| 8 | Bacteroides fragilis |
| 9 | Bordetella avium |
| 10 | Bordetella bronchiseptica (ATCC 19395)4,5,10 |
| 11 | Bordetella bronchiseptica (ATCC 4617)4,6,10 |
| 12 | Bordetella bronchiseptica (ATCC BAA-588) |
| 13 | Bordetella bronchiseptica (Clinical Isolate) |
| 14 | Bordetella hinzii |
| 15 | Bordetella holmesii (F061) |
| 16 | Bordetella holmesii (C690) |
| 17 | Bordetella holmesii (ATCC 51541)9 |
| 18 | Bordetella holmesii (NCTC 13202) |
| 19 | Bordetella petrii4 |
| 20 | Bordetella trematum |
| 21 | Burkholderia cepacia |
| 22 | Candida albicans7,11 |
| 23 | Chlamydophila pneumoniae |
| 24 | Citrobacter freundii |
| 25 | Corynebacterium diptheriae |
| 26 | Coxsackievirus9 |
| 27 | Cytomegalovirus |
| 28 | Echovirus |
| 29 | Enterobacter aerogenes4 |
| 30 | Enterococcus faecalis |
| 31 | Enterovirus |
| 32 | Epstein-Barr virus |
| 37 | Human Coronavirus 229E |
| 38 | Human Coronavirus OC433 |
| 39 | Influenza A |
| 40 | Influenza B3 |
| 41 | Klebsiella pneumoniae4 |
| 42 | Lactobacillus acidophilus |
| 43 | Lactobacillus plantarum |
| 44 | Legionella pneumophila |
| 45 | Metapneumovirus hMPV 20 Type A2 |
| 46 | Metapneumovirus hMPV 8 Type B2 |
| 47 | Moraxella catarrhalis3 |
| 48 | Morganella morganii |
| 49 | Mumps virus |
| 50 | Mycobacterium tuberculosis |
| 51 | Mycoplasma pneumoniae |
| 52 | Neisseria elongata |
| 53 | Neisseria meningitidis |
| 54 | Oligella ureolytica |
| 55 | Parainfluenza Type 1 |
| 56 | Parainfluenza Type 2 |
| 57 | Parainfluenza Type 3 |
| 58 | Parvimonas micra1 |
| 59 | Proteus vulgaris3 |
| 60 | Pseudomonas aeruginosa9 |
| 61 | Ralstonia paucula2 |
| 62 | Respiratory Syncytial Virus |
| 63 | Rhinovirus |
| 64 | Staphyloccocus aureus |
| 65 | Staphyloccocus aureus (MRSA) |
| 66 | Staphylococcus epidermidis |
| 67 | Staphylococcus hominis |
| 68 | Stenotrophomonas maltophilia |
Table 11.11: Microorganism Information
Executive Summary
16
| Microorganism | |||
|---|---|---|---|
| 33 | Escherichia coli | 69 | Streptococcus pneumoniae |
| 34 | Fusobacterium necrophorum3 | 70 | Streptococcus pyogenes |
| 35 | Haemophilus influenzae | 71 | Streptococcus salivarius |
| 36 | Human Bocavirus | 72 | Bordetella pertussis8 |
| 73 | Bordetella parapertussis8 |
1 Formerly Micromonas micros and Peptostreptococcus micros
2 Formerly Cupriavidus pauculus
3 A false positive result was observed in 1 out of 6 replicates during cross-reactivity testing.
4 B. pertussis was detected in 5/6 replicates during microbial interference studies.
- ് B. parapertussis (BPP) was not detected in one of the three replicates of BPP+CRO when CRO was spiked at 1.35x10 °CFU/mL. However, B. parapertussis was detected in all replicates when the CRO concentration was reduced to 10° CFU/mL. All replicates of B. pertussis were detected when CRO was spiked at 1.35x108 CFU/mL.
- 7 A false positive for B. parapertussis was detected in one of the three replicates of BP+CRO when CRO was spiked at 8.7x10 CFU/mL. However, no false positive for B. parapertussis was detected when the CRO concentration was reduced to 10" CFU/mL.
- ็ High concentration of B. pertussis (≥ 10 cfu/mL) was tested with low concentration of B. parapertussis (3x LoD) and vice-versa to evaluate the impact of microbial interference in a co-infection setting, during cross-reactivity studies. B. pertussis and B. parapertussis were also tested in a low/low, high/high setting, where low concentration was 3x LOD and high concentration was 100x LOD.
- ి A total of 6 replicates were tested with 1 out of 6 replicates result for B. parapertussis, during microbial interference studies.
- 10 nitial testing of 3 replicates resulted in a false negative for B. pertussis, during microbial interference studies. Subsequent repeat testing yielded the same outcome. After concentration of the CRO was reduced to 10° CFU/mL, expected positivity was achieved.
- 11 Initial testing of 3 replicates resulted in a false positive for B. parapertussis, during microbial interference studies. Subsequent repeat testing yielded the same outcome. After concentration of the CRO was reduced to 10 °CFU/mL, expected positivity was achieved.
Carry-Over/Cross-Contamination:
Carry-over and cross contamination for the ARIES® Bordetella Assay was evaluated by using samples consisting of Bordetella pertussis culture diluted into pooled natural negative human nasopharyngeal swab matrix at a concentration of 1x10 CFU/mL. Testing was performed using thirty high positive Bordetella pertussis samples in series alternating with thirty Bordetella pertussis and Bordetella parapertussis negative (pooled natural negative human nasopharyngeal swab matrix) samples. The high positive samples were run adjacent to negative samples across five consecutive runs using one ARIES® System.
The result of the study was no carry-over or cross contamination observed.
- g. Assay cut-off:
6 B. pertussis (BP) was not detected in one of the three replicates of BP+CRO when CRO was spiked at 3.64x10° CFU/mL. However, B. pertussis was detected in all replicates when the CRO concentration was reduced to 10° CFU/mL. All replicates of B. parapertussis were detected when CRO was spiked at 3.64x10° CFU/mL.
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Each target in the ARIES® Bordetella assay (B. pertussis and B. parapertussis) has a Ct cut-off, Tm window, and Tm Peak Threshold. In addition, the internal sample process control (SPC) also has a corresponding Ct cut-off, Tm window, and Tm Peak Threshold. Collectively, the cut-off values compose the assay protocol file parameters, which are used to determine the assay result for the detection target as POSITIVE, NEGATIVE, or INVALID.
The Assay Protocol File parameters were determined, and their performance in the ARIES® Bordetella Assay were evaluated according to the following general procedure:
- Initial Assay Protocol File parameters were set during internal optimization studies
- The final Assay Protocol File parameters were then established during internal verification studies
- . The selected Assay Protocol File parameter values were utilized in the determination of assay performance in the multi-site clinical trial conducted for the ARIES® Bordetella Assay
-
- Comparison Studies:
- Method comparison with predicate device: a.
Not applicable.
- Collection Media Comparison: b.
The compatibility of four media, UTM™, M5 , M6™ and ESwab™, was evaluated with the ARIES® Bordetella Assay using two strains each of Bordetella pertussis and Bordetella parapertussis. Each strain of Bordetella was independently diluted in the media at a concentration of three times the limit of detection of the ARIES® Bordetella Assay for that strain. The study was performed in multiple assay runs with twenty replicates for each media type and each strain of Bordetella and three replicates for each media type by itself as the negative control.
The results of the study are shown in Table 11.12. Based on this study, the four different types of media when tested with the two different types of Bordetella strains were determined to be compatible with the ARIES® Bordetella Assay.
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| Concentration/
Target type | Media Type | Mean Ct ± SD
(cycles) | % Positivity/
Detected |
|-----------------------------------------|---------------------------------|--------------------------|---------------------------|
| 3x LoD B. pertussis
(A639) | M6 | 35.8 ± 0.96 | 100% (20/20) |
| | M5 | 35.6 ± 0.66 | 100% (20/20) |
| | Universal Transport Media (UTM) | 36.4 ± 0.86 | 100% (20/20) |
| | ESwab (Modified Liquid Amies) | 36.2 ± 0.75 | 100% (20/20) |
| 3x LoD B. pertussis
(BAA-589) | M6 | 35.9 ± 0.67 | 100% (20/20) |
| | M5 | 36.7 ± 1.03 | 100% (20/20) |
| | Universal Transport Media (UTM) | 37.0 ± 0.76 | 95% (19/20) |
| | ESwab (Modified Liquid Amies) | 36.0 ± 0.68 | 100% (20/20) |
| 3x LoD B.
parapertussis
(A747) | M6 | 33.8 ± 0.53 | 100% (20/20) |
| | M5 | 33.6 ± 0.79 | 100% (20/20) |
| | Universal Transport Media (UTM) | 34.9 ± 0.62 | 100% (20/20) |
| | ESwab (Modified Liquid Amies) | 34.9 ± 1.05 | 100% (20/20) |
| 3x LoD B.
parapertussis
(BAA-587) | M6 | 33.8 ± 0.58 | 100% (20/20) |
| | M5 | 33.0 ± 0.67 | 100% (20/20) |
| | Universal Transport Media (UTM) | 34.7 ± 0.83 | 100% (20/20) |
| | ESwab (Modified Liquid Amies) | 33.9 ± 0.54 | 100% (20/20) |
| aNo Target | M6 | a29.9 ± 0.20 | 100% (3/3) |
| | M5 | a30.0 ± 0.40 | 100% (3/3) |
| | Universal Transport Media (UTM) | a29.4 ± 0.36 | 100% (3/3) |
| | ESwab (Modified Liquid Amies) | a30.2 ± 0.06 | 100% (3/3) |
Table 11.12: ARIES® Bordetella Assay Media Equivalency Results
ే Data shown for DNA SPC
ﻥ Swab Comparison:
The compatibility of 3 different types of nasopharyngeal swabs, Flocked, Rayon and Polyester was evaluated with the ARIES® Bordetella Assay using one strain each of Bordetella pertussis and Bordetella parapertussis. The Bordetella strains were diluted independently into pooled natural negative human nasopharyngeal swab matrix and applied to each swab type. The collection swabs were then inserted and broken off inside 3mL Universal Transport Medium (UTM) vials for a final concentration of 3x LoD in the transport medium. In addition, a negative sample consisting of the swab only transferred in UTM was also evaluated.
The results of the study are shown in Table 11.13. Based on this study, the three different types of nasopharyngeal swabs when tested with the Bordetella strains were determined to be compatible with the ARIES® Bordetella Assay.
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| Concentration/ Target
type | Swab Type | Mean Ct ± SD (cycles) | % Positivity/
Detected |
|-----------------------------------|----------------------------|-----------------------|---------------------------|
| 3x LoD B. pertussis
(A639) | Flocked Swab | 38.4 ± 1.33 | 100% (9/9) |
| | Polyester Swab | 36.9 ± 0.75 | 100% (9/9) |
| | Rayon Swab | 37.3 ± 1.53 | 100% (9/9) |
| | No Swab (Positive Control) | 37.6 ± 1.04 | 100% (3/3) |
| 3x LoD B. parapertussis
(A747) | Flocked Swab | 35.4 ± 0.87 | 100% (9/9) |
| | Polyester Swab | 33.8 ± 0.92 | 100% (9/9) |
| | Rayon Swab | 34.1 ± 1.18 | 100% (9/9) |
| | No Swab (Positive Control) | 34.2 ± 0.90 | 100% (3/3) |
| aNo Target | Flocked Swab | a31.8 ± 0.45 | 100% (3/3) |
| | Polyester Swab | a31.1 ± 0.21 | 100% (3/3) |
| | Rayon Swab | a31.3 ± 0.42 | 100% (3/3) |
Table 11.13: ARIES® Bordetella Assay Nasopharyngeal Swab Equivalency Results
ే Data shown for DNA SPC
3. Clinical Performance:
The clinical performance of ARIES® Bordetella Assay was evaluated using leftover deidentified nasopharyngeal swab (NPS) specimens prospectively collected from pediatric and adult patients suspected of having respiratory tract infection attributable to B. pertussis or B. parapertussis.
Five (5) geographically distinct clinical sites within the United States prospectively collected specimens from July through November 2016. The clinical performance of the ARIES Bordetella Assay was evaluated on these prospectively collected specimens at four (4) of the five (5) clinical sites using the ARIES System. One (1) clinical site collected specimens then shipped them frozen on dry ice to one of the other clinical sites for ARIES Bordetella Assay testing. Specimens included in the clinical study consisted of leftover de-identified nasopharyngeal swabs (NPS) specimens prospectively collected from pediatric and adult patients suspected of having respiratory tract infection attributable to B. pertussis or B. parapertussis.
A total of 1052 unique specimens that met the pre-determined inclusion criteria were included in the study and tested for B. pertussis or B. parapertussis by both the reference method and the ARIES "Bordetella Assay. The performance of ARIES" Bordetella Assay for B. pertussis and B. parapertussis was compared to a composite comparator assay consisting of two well-characterized real-time PCR assays (for each bacterial pathogen) followed by confirmation of positive PCR amplification product with bi-directional sequencing. Comparator PCR assays for B. pertussis and B. parapertussis targeted unique sequences within the promoter region of the ptxA gene and IS1001 insertion region (respectively) that were different than those targeted by the ARIES Bordetella Assay. Specimens were characterized as positive for B. pertussis or B. parapertussis if one out of two comparator PCR assays was positive (Ct values ≤40) and confirmed by bi-directional sequencing, or if
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both comparator PCR assays were positive. Specimens were characterized as B. pertussis or B. parapertussis negative if one out of two comparator PCR assays was negative (Ct values >40) and confirmed by bi-directional sequencing, or if both comparator PCR assays were negative.
Comparator real-time PCR and bi-directional sequencing assays were performed at a centralized testing facility. Clinical runs and re-runs using ARIES Bordetella Assay were carried out by trained operators at the four (4) testing sites on specimens that were either kept refrigerated at 2°C to 8°C for up to 72 hours prior to testing (N=667; 63.4%) or stored frozen at -65°C to -95°C for up to 12 days prior to testing (N=385; 36.6%).
Out of the 1052 clinical specimens included in the prospective study analysis, 1043 (99.1%) generated valid ARIES Bordetella Assay results (i.e., positive or negative) on the first attempt. There were 9 specimens (9/1052; 0.9%) that were re-tested with ARIES Bordetella Assay because they yielded invalid results in the initial run (N=3) or because of instrument error (N=6). All nine (9) specimens generated valid ARIES results upon repeat testing.
In the prospective study, the ARIES Bordetella Assay Positive Percent Agreement (PPA) for B. pertussis was reported to be 93.8% (30/32) with a lower bound of the 95% confidence interval of 79.2%. Positive Percent Agreement of the ARIES Bordetella Assay for B. parapertussis was reported to be 100% (2/2) with a lower bound of the 95% confidence interval of 15.8%. Negative Percent Agreement of the ARIES Bordetella Assay for B. pertussis and B. parapertussis were respectively; 98.9% (1009/1020) with a lower bound 95% confidence interval of 98.1%, and 99.8% (1048/1050) with a lower bound 95% confidence interval of 99.3%.
Due to the low prevalence B. pertussis and B. parapertussis observed in the prospective study, the clinical sample set was supplemented with banked (pre-selected) B. pertussis (N=37) and B. parapertussis (N=20) positive specimens as well as contrived B. parapertussis specimens (N=50). Pre-selected B. pertussis specimens were collected at six (6) clinical sites in the United States while pre-selected B. parapertussis specimens were collected at three (3) sites also located in the United States. Contrived B. parapertussis samples were prepared by spiking well-characterized bacterial strains into individual negative clinical samples (NP swabs) at clinically relevant titers. B. parapertussis contrived specimens were prepared at analyte concentrations near the ARIES Bordetella Assay limit of detection (LoD) as well as concentrations spanning the clinically moderate to high positive ranges. The presence of the expected bacterial target in each of the pre-selected and contrived specimens was confirmed by comparator real-time PCR and bi-directional sequencing assays. In addition, bacterial organism concentrations in contrived specimens were verified by culture methods (plating and colony count). In order to minimize bias, pre-selected and contrived specimens were tested along with an equal number of unique negative clinical specimens in a randomized, blinded fashion at three (3) external testing sites. ARIES Bordetella Assay accurately detected all 37 B. pertussis (100% PPA; 95% confidence interval: 90.5% - 100%) and all 20 B. parapertussis positive specimens tested (100% PPA; 95% confidence interval: 83.2% - 100%). One of the pre-selected specimens generated a false
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positive result for B. parapertussis by ARIES® Bordetella Assay when compared to the composite comparator method (01-122). All fifty (50) contrived B. parapertussis samples were accurately detected by the ARIES Bordetella assay (100% PPA; 95% confidence interval 92.9%- 100%). All but one of the pre-selected and contrived specimens generated valid ARIES result upon initial testing. The invalid result was resolved upon re-test.
The performance of the ARIES Bordetella Assay for B. pertussis and B. parapertussis as compared to the composite reference method is summarized in the Table 11.14 and Table 11.15 for prospective, pre-selected and contrived specimens. The overall performance of the ARIES Bordetella Assay in combined prospective, pre-selected and contrived specimens is also presented.
Specimen Description PPA 95% Cl NPA 95% Cl 30/321 Prospective 93.8% 79.2% - 99.2% 1009/1020 98.9% 98.1% - 99.5% Pre-selected 37/37 100% 90.5% - 100% 77/77 100% 95.3% - 100% 67/69 97.1% 89.9% - 99.6% 1086/1097 99.0% 98.2% - 99.5% Total
Table 11.14: ARIES® Bordetella Assay Performance for B. pertussis
4 Two (2) prospective specimens generated false negative results by ARIES® Bordetella assay when comparite comparator method (02-179 and 06-267).
| Table 11.15: ARIES® Bordetella Assay Performance for B. parapertussis | ||
|---|---|---|
| ------------------------------------------------------------------------ | -- | -- |
| Specimen Description | PPA | 95% CI | NPA | 95% CI | ||
|---|---|---|---|---|---|---|
| Prospective | 2/2 | 100% | 15.8% - 100% | 1048/1050 | 99.8% | 99.3% - 100% |
| Pre-selected | 20/20 | 100% | 83.2% - 100%) | 93/941 | 98.9% | 94.2% - 100% |
| Contrived | 50/50 | 100% | 92.9% - 100% | 50/50 | 100% | 92.9% - 100% |
| Total | 72/72 | 100% | 95.0% - 100% | 1191/1194 | 99.7% | 99.3% - 99.9% |
් One (1) pre-selected specimen generated a false positive result by ARIES® Bordetella Assay when compared to the composite comparator method (01-122).
The study results demonstrate that the diagnostic accuracy of ARIES® Bordetella Assay is acceptable for the safe and effective detection of B. pertussis and B. parapertussis in nasopharyngeal swabs (NPS) specimens from patients suspected of having respiratory tract infection attributable to B. pertussis or B. parapertussis.
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4. Expected values:
The overall prevalence of B. pertussis and B. parapertussis, as reported by the ARIES Bordetella Assay, in prospectively collected symptomatic clinical specimens during the enrollment period was 3.9% (41/1052) and 0.4% (4/1052) respectively.
N. Proposed Labeling:
The labeling provided in the submission satisfies the requirements of 21 CFR 809.10.
O. Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.