(33 days)
Not Found
No
The device description and performance studies focus on a laboratory-based nucleic acid amplification test (HDA) with manual evaluation of results using a lateral flow strip. There is no mention of AI or ML in the document.
No.
The AmpliVue® Bordetella Assay is an in vitro diagnostic test designed to detect Bordetella pertussis nucleic acids for diagnostic purposes, not for treating or preventing disease.
Yes
The "Intended Use / Indications for Use" section explicitly states that the "AmpliVue® Bordetella Assay is an in vitro diagnostic test for the qualitative detection of Bordetella pertussis nucleic acids..." and "aid in diagnosis infection."
No
The device description clearly outlines a physical, self-contained disposable amplification device and a detection chamber, indicating it is a hardware-based in vitro diagnostic test, not software only.
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 AmpliVue® Bordetella Assay is an "in vitro diagnostic test".
- Purpose: The test is designed for the qualitative detection of Bordetella pertussis nucleic acids isolated from patient specimens (nasopharyngeal swabs) to aid in the diagnosis of respiratory tract infection. This is a classic definition of an in vitro diagnostic device.
- Specimen Type: It uses biological specimens (nasopharyngeal swabs) taken from the human body.
- Testing Location: It is intended for use in laboratory settings (hospital, reference, or state laboratories), which is typical for IVDs.
- Performance Studies: The document includes detailed performance studies (Clinical Sensitivity, Reproducibility, Analytical Sensitivity, Analytical Specificity, etc.) which are required for the regulatory approval of IVDs.
- Predicate Device: A predicate device (K133673; illumigene® Pertussis DNA Amplification Assay) is listed, which is common in the regulatory submission process for IVDs.
N/A
Intended Use / Indications for Use
The AmpliVue® Bordetella Assay is an in vitro diagnostic test for the qualitative detection of Bordetella pertussis nucleic acids isolated from nasopharyngeal swab specimens obtained from patients suspected of having respiratory tract infection attributable to Bordetella pertussis.
The AmpliVue® Bordetella Assay utilizes helicase-dependent amplification (HDA) of the insertion sequence IS481 and a self-contained disposable amplification detection device that allows for manual evaluation of assay results. The IS481 sequence can also be found in strains of other organisms (i.e., B. holmesii and B. bronchiseptica). 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 AmpliVue® Bordetella Assay do not preclude B. pertussis infection and positive results do not rule out co-infection with other respiratory pathogens. Results from the AmpliVue® Bordetella Assay should be used in conjunction with information obtained during the patient's clinical evaluation as an aid in diagnosis of Bordetella pertussis infection and should not be used as the sole basis for treatment or other patient management decisions.
The AmpliVue Bordetella Assay is intended for use in hospital, reference or state laboratory settings. The device is not intended for point-of-care use.
Product codes
OZZ
Device Description
The AmpliVue® Bordetella Assay combines simple processing, an isothermal amplification technology named helicase-dependent amplification (HDA), and a selfcontained disposable amplicon detection device, for the detection of Bordetella pertussis from nasopharyngeal swabs.
Patient samples are collected using a nasopharyngeal swab and placed into a liguid medium. Fifty microliters (50 µL) of the sample are then transferred to a process buffer that is provided with the kit and mixed. The Process Buffer tubes are heated at 95 °C for 10 minutes. Fifty microliters (50 µL) of the Process Buffer containing sample is added to a reaction tube containing lyophilized mix of HDA reagents. Included in the reaction mix are the isothermal polymerase, helicase and single stranded binding protein. After completion of the HDA reaction the reaction tube is transferred to the amplicon cartridge containing the running buffer. The amplicon cartridge is closed and inserted into the detection chamber. The detection chamber is activated by depressing the detection chamber handle. Upon activation, the reservoir containing the running buffer and the 0.2 mL tube containing the amplicon is punctured and the solutions are wicked to the lateral flow strip.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
nasopharyngeal swab speciments
Indicated Patient Age Range
Not Found
Intended User / Care Setting
hospital, reference or state laboratory settings. The device is not intended for point-of-care use.
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
Not Found
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Reproducibility: A blinded and randomized study panel containing Bordetella pertussis (BP) negative and positive samples (5x and 2x,) were tested at three (3) test sites (one in-house laboratory and two (2) clinical sites). Each site tested a reproducibility panel and Assay Controls for five (5) days in triplicate. Testing was done by two operators at each site. Each operator ran the panel once a day using one lot of AmpliVue Bordetella Assay. A total of four hundred fifty (450) specimens were tested (including controls). The AmpliVue Bordetella Assay generated reproducible results in this study.
- Key Results:
- BP Low Positive: Overall Percent Agreement 100% (95% Confidence Interval 95.9% to 100%)
- BP Moderate Positive: Overall Percent Agreement 100% (95% Confidence Interval 95.9% to 100%)
- BP Negative: Overall Percent Agreement 100% (95% Confidence Interval 95.9% to 100%)
- BP Positive Control: Overall Percent Agreement 100% (95% Confidence Interval 95.9% to 100%)
- BP Negative Control: Overall Percent Agreement 100% (95% Confidence Interval 95.9% to 100%)
Detection limit: The analytical sensitivity (limit of detection or LoD) of the AmpliVue "Bordetella Assay was determined using quantified (CFU/mL) cultures of two (2) Bordetella pertussis bacterial stocks, BP A639 and E431 serially diluted in negative nasal matrix. The LoD is defined as the lowest concentration at which 95% of all replicates tested positive.
- Bacterial Strain A639: 2,358 CFU/mL (3.93 CFU/Assay)
- Bacterial Strain E431: 761 CFU/mL (1.27 CFU/Assay)
- Key Result: The assay LOD for Bordetella pertussis is 3.93 CFU/assay or 2,358 CFU/mL (sample input).
Analytical Specificity (Cross Reactivity): A study was performed to evaluate the cross-reactivity of the AmpliVue "Bordetella Assay with seventy-nine (79) other microorganisms potentially found in specimens collected to test for Bordetella pertussis (BP) infection. Cross-reactive microorganism was tested at clinically relevant levels of viruses (10 fpfu/mL) and bacteria (10 cfu/mL) in the device.
- Key Results: The study determined that 1 of 4 Bordetella bronchiseptica strains (strain 4617) and 4 of 4 Bordetella holmesii strains tested were cross-reactive with the AmpliVue Bordetella Assay. These results are expected as 5% of all Bordetella bronchiseptica strains and all Bordetella holmesii strains are known to carry the IS481 target sequence.
Interference: A study was conducted to determine if the AmpliVue "Bordetella assay is inhibited in the presence of a panel of seventeen (17) substances potentially present in specimens collected to test for Bordetella pertussis infection. Each of the potential interfering substances was tested in three replicates in the presence and absence of near LOD (2x) levels of B. pertussis in the AmpliVue Bordetella Assay. Substances were introduced into the assay at concentrations which were medically relevant.
- Key Results: There was no evidence of interference caused by the substances tested.
Analytical Reactivity (Inclusivity): The reactivity of the AmpliVue " Bordetella assay was evaluated against an additional six (6) strains of Bordetella pertussis at concentrations near the level of detection (LoD) of the assay. Each strain was tested as three replicates.
- Key Results: All seven strains were detected by the AmpliVue 'Bordetella Assay.
Matrix Comparison: The compatibility of eight different types of media with the AmpliVue Bordetella assay was determined using Validation Lot reagents and the proposed workflow. Four strains of pre-titered Bordetella pertussis stocks were tested at 1x LoD in the presence of eight different types of media.
- Key Results: For Bordetella pertussis, 4 of 4 strains (A639, E431, BAA 1335, and 51445) were determined to be compatible with the 8 different media types when tested at the assay LoD.
Clinical Sensitivity/Specificity: Performance characteristics of the AmpliVue® Bordetella Assay was established in the Spring to Summer of 2014 (April to August 2014) at four locations in the United States. Eight hundred forty two (842) fresh nasopharyngeal swab specimens were obtained from female and male patients suspected of having respiratory tract infection attributable to Bordetella pertussis from were collected and transported to each laboratory for testing with the AmpliVue® Bordetella Assay. Clinical performance was based on comparison of the AmpliVue Bordetella Assay results to those obtained by Composite Reference Method that included two manufacturer validated, IS481-targeted real-time PCR assays (PCR1 and PCR2) followed by bi-directional sequencing from PCR positive specimens. Six (6) specimens (0.7%) were invalid and removed from analysis, leaving 836 specimens.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
- Positive Percent Agreement: 97.0% (64/66) (95% CI: 89.6% to 99.2%)
- Negative Percent Agreement: 98.1% (755/770) (95% CI: 96.8% to 98.8%)
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.
0
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
QUIDEL CORPORATION RONALD LOLLAR SENIOR DIRECTOR, CLINICAL AND REGULATORY AFFAIRS 2005 EAST STATE STREET SUITE 100 ATHENS OH 45701
December 10, 2014
Re: K143206
Trade/Device Name: Amplivue Bordetella Assay Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory viral panel multiplex nucleic acid assay Regulatory Class: II Product Code: OZZ Dated: November 6, 2014 Received: November 7, 2014
Dear Dr. Lollar:
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 Parts 801 and 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 (OS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
1
If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 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/Resourcesfor You/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 yours,
Uwe Scherf -S for
Sally Hojvat, 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) K143206
Device Name Amplivue Bordetella Assay
Indications for Use (Describe)
The AmpliVue® Bordetella Assay is an in vitro diagnostic test for the qualitative detection of Bordetella pertussis nucleic acids isolated from nasopharyngeal swab speciments suspected of having respiratory tract infection attributable to Bordetella pertussis.
The AmpliVue® Bordetella Assay utilizes helicase-dependent amplification (HDA) of the insertion sequence IS481 and a selfcontained disposable amplification device that allows for manual evaluation of assay results. The IS48 sequence can also be found in strains of other organisms (i.e., 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 AmpliVue® Bordetella Assay do not prection and positive results do not rule out coinfection with other respiratory pathogens. Results from the AmpliVue® Bordetella Assay should be used in conjunction with information obtained during the patient's clinical evaluation as an aid in diagnosis infection and should not be used as the sole basis for treatment or other patient management decisions.
The AmpliVue® Bordetella Assay is intended for use in hospital, reference or state laboratory settings. The device is not intended for point-of-care use.
Type of Use (Select one or both, as applicable)
| ☑ Prescription Use (Part 21 CER 801 Subpart D) |
|---|
| ☐ Over-The-Counter Use (21 CER 801 Subpart C) |
PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON A SEPARATE PAGE IF NEEDED.
FOR FDA USE ONLY
Concurrence of Center for Devices and Radiological Health (CDRH) (Signature)
3
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4
Applicant:
Quidel Corporation 12544 High Bluff Drive, Suite 200 San Diego, California 92130 Telephone: 858-552-7910 Fax: 858-646-8045
Contact Information:
Ronald H. Lollar, Senior Director Clinical and Regulatory Affairs 2005 East State Street, Suite 100 Athens, Ohio 45701 740-589-3300 – Corporate number 740-589-3373 – Desk phone 858-552-6451—Fax Ron.Lollar@quidel.com
Date of preparation of 510(k) summary:
November 05, 2014
A. 510(k) Number:
B. Purpose for Submission:
To obtain substantial equivalence for the AmpliVue Bordetella Assay
C. Measurand:
Insertion sequence IS481 of Bordetella pertussis
D. Type of Test:
Helicase-dependent amplification (HDA)
5
E. Applicant:
Quidel Corporation
F. Proprietary and Established Names:
AmpliVue Bordetella Assay
G. Regulatory Information:
| Product Code | Classification | Regulation Section | Panel |
|---|---|---|---|
| OZZ – Bordetella | |||
| pertussis DNA | |||
| assay system | Class 2 | 21 CFR 866.3980 – Respiratory | |
| viral panel multiplex nucleic acid | |||
| assay | Microbiology | ||
| (83) |
H. Intended Use:
1. Intended Use(s):
The AmpliVue® Bordetella Assay is an in vitro diagnostic test for the qualitative detection of Bordetella pertussis nucleic acids isolated from nasopharyngeal swab specimens obtained from patients suspected of having respiratory tract infection attributable to Bordetella pertussis.
The AmpliVue® Bordetella Assay utilizes helicase-dependent amplification (HDA) of the insertion sequence IS481 and a self-contained disposable amplification detection device that allows for manual evaluation of assay results. The IS481 sequence can also be found in strains of other organisms (i.e., B. holmesii and B. bronchiseptica). 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.
6
Negative results for the AmpliVue® Bordetella Assay do not preclude B. pertussis infection and positive results do not rule out co-infection with other respiratory pathogens. Results from the AmpliVue® Bordetella Assay should be used in conjunction with information obtained during the patient's clinical evaluation as an aid in diagnosis of Bordetella pertussis infection and should not be used as the sole basis for treatment or other patient management decisions.
The AmpliVue Bordetella Assay is intended for use in hospital, reference or state laboratory settings. The device is not intended for point-of-care use.
2. Indication(s) for Use:
Same as Intended Use.
3. Special conditions for use statement(s):
- For in vitro diagnostic use only
- For prescription use only
4. Special instrument requirements:
None
l. Device Description:
The AmpliVue® Bordetella Assay combines simple processing, an isothermal amplification technology named helicase-dependent amplification (HDA), and a selfcontained disposable amplicon detection device, for the detection of Bordetella pertussis from nasopharyngeal swabs.
Patient samples are collected using a nasopharyngeal swab and placed into a liguid medium. Fifty microliters (50 µL) of the sample are then transferred to a process buffer that is provided with the kit and mixed. The Process Buffer tubes are heated at 95 °C for 10 minutes. Fifty microliters (50 µL) of the Process Buffer containing sample is added to a reaction tube containing lyophilized mix of HDA reagents. Included in the reaction mix are the isothermal polymerase, helicase and single stranded binding protein. After completion of the HDA reaction the reaction tube is transferred to the amplicon cartridge containing the running buffer. The amplicon cartridge is closed and inserted into the detection chamber. The detection chamber is activated by depressing the detection chamber
7
handle. Upon activation, the reservoir containing the running buffer and the 0.2 mL tube containing the amplicon is punctured and the solutions are wicked to the lateral flow strip.
Materials Provided:
- . 16 Tests per Kit
| Symbol | Component | Quantity | Storage |
|---|---|---|---|
| 1 | Detection Cassettes | 16/kit | 2° to 30°C |
| 2 | Process Buffer | 16 tubes/kit 1.45mL | 2° to 30°C |
| 3 | Reaction Tubes | 16 tubes/kit | 2° to 8°C |
| 4 | Amplicon Cartridge | 16/kit | 2° to 30°C |
Materials required but not provided:
- . External controls for Bordetella pertussis (e.g. Quidel Molecular Bordetella Control Set #M117, which contains positive and negative controls, serves as an external processing and extraction control)
- Sterile DNAse-free filter-blocked or positive displacement micropipettor tips
- Micropipettor ●
- Stopwatch or timer
- Scissors or a blade ●
- Micro tube tray
- Heat block capable of 95° C ± 2° C temperature ●
- Heat block with heated lid capable of 64° ± 2° C temperature
- . Thermometer
Substantial Equivalence Information: J.
-
- Predicate device name(s):
illumigene® Pertussis DNA Amplification Assay
- Predicate device name(s):
-
- Predicate 510(k) number(s): K133673
8
3. Comparison with predicate:
| Similarities | ||
|---|---|---|
| Item | AmpliVue® Bordetella Assay | illumigene® Pertussis DNA |
| Amplification Assay | ||
| (K133673) | ||
| Intended Use | The AmpliVue® Bordetella Assay is an | |
| in vitro diagnostic test for the | ||
| qualitative detection of Bordetella pertussis nucleic acids isolated from | ||
| nasopharyngeal swab specimens | ||
| obtained from patients suspected of | ||
| having respiratory tract infection | ||
| attributable to Bordetella pertussis . |
The AmpliVue® Bordetella Assay
utilizes helicase-dependent
amplification (HDA) of the insertion
sequence IS481 and a self-contained
disposable amplification detection
device that allows for manual
evaluation of assay results. The IS481
sequence can also be found in strains
of other organisms (i.e., B. holmesii
and B. bronchiseptica ). 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 | 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 |
| Similarities | | |
| Item | AmpliVue® Bordetella Assay | illumigene® Pertussis DNA
Amplification Assay
(K133673) |
| | 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 AmpliVue®
Bordetella Assay do not preclude B. pertussis infection and positive results
do not rule out co-infection with other
respiratory pathogens. Results from
the AmpliVue® Bordetella Assay should
be used in conjunction with
information obtained during the
patient's clinical evaluation as an aid in
diagnosis of Bordetella pertussis
infection and should not be used as
the sole basis for treatment or other
patient management decisions. | 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 |
| Similarities | | |
| Item | AmpliVue® Bordetella Assay | illumigene® Pertussis DNA
Amplification Assay
(K133673) |
| | | 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
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 |
| Similarities | | |
| Item | AmpliVue® Bordetella Assay | illumigene® Pertussis DNA
Amplification Assay
(K133673) |
| | | state laboratory settings.
The device is not
intended for point-of-
care use. |
| Sample Types | Nasopharyngeal swab specimens
obtained from patients suspected of
having respiratory tract infection
attributable to Bordetella pertussis | Same |
| Sample Heat
Lysis | Manual | Same |
| Target
Sequence
Detected | Bordetella pertussis IS481 insertional
element | Same |
9
AmpliVue® Bordetella Assay 11/05/2014 Page 6 of 19
10
11
| Differences | ||
|---|---|---|
| Item | AmpliVue ® Bordetella Assay | illumigene® Pertussis DNA |
| Amplification Assay | ||
| (K133673) | ||
| DNA Amplification | ||
| Technology | Helicase-dependent | |
| amplification (HDA); self- | ||
| contained | Loop-Mediated Isothermal | |
| Amplification (LAMP); self- | ||
| contained and automated | ||
| Self-Contained | ||
| System Assay after | ||
| sample preparation | No | Yes |
| Detection Technique | Manual | Automated |
| Instrument | None | illumipro-10™ |
12
| Differences | ||
|---|---|---|
| Item | AmpliVue® Bordetella Assay | illumigene® Pertussis DNA |
| Amplification Assay | ||
| (K133673) | ||
| Testing Time | 85 - 90 minutes | 60 - 70 minutes |
K. Standard/Guidance Document Referenced (if applicable):
Not applicable
L. Test Principle:
The AmpliVue® Bordetella Assay detects Bordetella pertussis DNA isolated from nasopharyngeal swab specimens obtained from symptomatic patients suspected of having respiratory tract infection attributable to Bordetella pertussis. The assay consists of three major steps: 1) specimen preparation, 2) isothermal Helicase-Dependent Amplification (HDA) of a target sequence of B. pertussis, and 3) detection of the amplified DNA by targetspecific hybridization probes via a colorimetric reaction on a lateral-flow strip which is embedded in a self-contained disposable cassette to prevent amplicon contamination.
Patient samples are collected using a nasopharyngeal swab and placed into a liquid medium. Fifty microliters (50 µL) of the sample are then transferred to a process buffer that is provided with the kit and mixed. The Process Buffer tubes are heated at 95 °C for 10 minutes. Fifty microliters (50 µL) of the Process Buffer containing sample is added to a reaction tube containing lyophilized mix of HDA reagents.
A HDA reaction is carried out in the Reaction Tube which contains lyophilized HDA reagents, dNTPs, primers and probes. Incubation at 64°C for 60 minutes results in isothermal amplification of the target sequence by B. pertussis specific primers. The amplified DNA is detected by a set of specific detection probes included in the Reaction Tube: B. pertussis target hybridizes to two specific probes labeled with Biotin (BioTEG) and 6-carboxyfluorescein (FAM). A competitive process control (PRC) is included in the Lysis Tube to monitor specimen processing and inhibitory substances in clinical samples, reagent failure or device failure. The PRC target is amplified and hybridizes to the PRC specific probes labeled with Biotin (BioTEG) and 2,4-dinitrophenyl (DNP).
Following completion of the HDA reaction, the Reaction Tube is transferred to a proprietary Cassette for detection. The Cassette is comprised of two components: 1) an
13
Amplicon Cartridge that holds the running buffer and the 0.2 mL Reaction Tube and 2) the Detection Chamber which houses the Amplicon Cartridge and an embedded vertical-flow DNA detection strip. The DNA detection strip is coated with anti-FAM and anti-DNP antibodies. Once the Cassette is closed, a razor blade and plastic pin located at the bottom of the Cassette opens the Reaction Tube and running buffer bulb, resulting in the release of their contents. The contents flows through a fiberglass paper connected to the DNA detection strip that is attached to a fiberglass pad pre-loaded with streptavidin-conjugated color particles. The B. pertussis amplicon with biotin- and FAM-labeled probes is captured by the anti-FAM antibodies at the test (T2) line, and the Process Control amplicon with biotin- and DNP-labeled probes is captured by the anti-DNP antibodies at the control (C) line. The streptavidin-conjugated color particles bind to the biotin in the probe-amplicon hybrid and the test results are displayed in the Cassette window as colored T2 and/or C lines that are visible to the naked eye.
Detection of B. pertussis is reported when the T2 line is visible through the detection window of the Cassette. No detection of B. pertussis is reported when only the C line is displayed. The assay is regarded as invalid when none of the lines are displayed.
M. Performance Characteristics:
1. Analytical performance:
- a. Precision/Reproducibility:
Reproducibility
In order to confirm the reproducibility of the AmpliVue Bordetella Assay a blinded and randomized study panel containing Bordetella pertussis (BP) negative and positive samples (5x and 2x,) were tested at three (3) test sites (one in-house laboratory and two (2) clinical sites). Each site tested a reproducibility panel and Assay Controls for five (5) days in triplicate. Testing was done by two operators at each site. Each operator ran the panel once a day using one lot of AmpliVue Bordetella Assay. A total of four hundred fifty (450) specimens were tested (including controls). The AmpliVue Bordetella Assay generated reproducible results in this study.
| SITE | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Category | Site #1 | Site #2 | Site #3 | Overall Percent | |||||
| Agreement | 95% | ||||||||
| Confidence | |||||||||
| Interval | |||||||||
| #expected | |||||||||
| results/# | |||||||||
| tested | % | ||||||||
| Agreement | #expected | ||||||||
| results/# | |||||||||
| tested | % | ||||||||
| Agreement | #expected | ||||||||
| results/# | |||||||||
| tested | % | ||||||||
| Agreement | |||||||||
| BP Low Positive | 30/30 | 100% | 30/30 | 100% | 30/30 | 100% | 90/90 | 100% | 95.9% to |
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AmpliVue Bordetella Assay 11/05/2014 Page 11 of 19
510(k) Summary
| SITE | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Site #1 | Site #2 | Site #3 | Overall Percent | 95% | |||||
| Category | #expected | ||||||||
| results/# | |||||||||
| tested | % | ||||||||
| Agreement | #expected | ||||||||
| results/# | |||||||||
| tested | % | ||||||||
| Agreement | #expected | ||||||||
| results/# | |||||||||
| tested | % | ||||||||
| Agreement | Agreement | Confidence | |||||||
| Interval | |||||||||
| (4,716 cfu/mL) | 100% | ||||||||
| BP Moderate Positive | |||||||||
| (11,790 cfu/mL) | 30/30 | 100% | 30/30 | 100% | 30/30 | 100% | 90/90 | 100% | 95.9% to |
| 100% | |||||||||
| BP Negative | 30/30 | 100% | 30/30 | 100% | 30/30 | 100% | 90/90 | 100% | 95.9% to |
| 100% | |||||||||
| BP Positive Control | 30/30 | 100% | 30/30 | 100% | 30/30 | 100% | 90/90 | 100% | 95.9% to |
| 100% | |||||||||
| BP Negative Control | 30/30 | 100% | 30/30 | 100% | 30/30 | 100% | 90/90 | 100% | 95.9% to |
| 100% |
The results suggest that there are no significant differences between different users and different sites on different days. Reproducibility studies are acceptable.
-
b. Linearity/assay reportable range:
Not applicable – This assay is qualitative. -
c. Traceability, Stability, Expected values (controls, calibrators, or methods):
Traceability:
Not applicable. This assay is qualitative.
Specimen Stability:
A study was performed to determine the specimen stability using a contrived sample at 2x LOD. The contrived sample was stored at 2°C to 8°C for varying lengths of time (24, 48, 72 and 96-hours). The samples were brought to room temperature and tested with the AmpliVue Bordetella Assay.
Based on this study the contrived 2x LOD sample was stable when stored at 2℃ to 8°C
Controls:
15
Controls (Quidel Molecular Bordetella Control Set #M117, which contains positive and negative controls, serves as an external processing and extraction control) were run on the AmpliVue Bordetella Assay each day of testing. These controls are described as follows:
- a. The process control is used to monitor sample processing, to detect HDA inhibitory specimens and to confirm the integrity of assay reagents and cassette detection. The process control is included in the Lysis Buffer tube.
- b. The external positive control may be treated as a patient specimen. The control should be sampled and tested as if it were a swab specimen and processed as described in the Assay Procedure. The external positive control is intended to monitor substantial reagent and cassette failure.
- The external negative control may be treated as a patient specimen. The ﻥ control should be sampled and tested as if it were a swab specimen and processed as described in the Assay Procedure. The external negative control is used to detect reagent or environmental contamination (or carry-over) by B. pertussis DNA or amplicon.
d. Detection limit:
The analytical sensitivity (limit of detection or LoD) of the AmpliVue "Bordetella Assay was determined using quantified (CFU/mL) cultures of two (2) Bordetella pertussis bacterial stocks, BP A639 and E431 serially diluted in negative nasal matrix. The LoD is defined as the lowest concentration at which 95% of all replicates tested positive.
The bacterial strains were freshly grown. The cell density of these bacterial suspensions was estimated using the OD600 reading. After a cell suspension of 0.25 McFarland units was established, the bacteria were serially diluted in PBS to densities ranging from 3x to 0.3x LoD levels based on preliminary studies.
Each test dilution was run as 20 replicates in the AmpliVue assay. The highest dilution where at least 19 of 20 replicates show detection of B. pertussis (95% positivity) was assigned the Limit of Detection of the strain. The CFU/mL was calculated based on the average bacterial plate count of the dilution.
16
510(k) Summary
| Bacterial
Strain | Concentration
CFU/mL | Concentration
CFU/Assay |
|---------------------|-------------------------|----------------------------|
| A639 | 2,358 | 3.93 |
| E431 | 761 | 1.27 |
The assay LOD for Bordetella pertussis is 3.93 CFU/assay or 2,358 CFU/mL (sample input).
e. Analytical specificity:
Cross Reactivity:
A study was performed to evaluate the cross-reactivity of the AmpliVue "Bordetella Assay with seventy-nine (79) other microorganisms potentially found in specimens collected to test for Bordetella pertussis (BP) infection. Cross-reactive microorganism was tested at clinically relevant levels of viruses (10 fpfu/mL) and bacteria (10 cfu/mL) in the device. The organisms and their concentrations included in the cross-reactivity study are shown in the table below.
| Organism | Test Concentration |
|---|---|
| Acinetobacter baumanii | 2.90 x106 cfu/mL |
| Arcanobacterium haemolyticum | 1.15 x106 cfu/mL |
| Bacteroides fragilis | 1.19 x106 cfu/mL |
| Bordetella avium | 3.85 x106 cfu/mL |
| Bordetella bronchiseptica | 9.45 x106 cfu/mL |
| Bordetella bronchiseptica | 1.17 x106 cfu/mL |
| Bordetella bronchiseptica (ATCC 4617) | 7.74 x106 cfu/mL |
| Bordetella bronchiseptica | 1.97 x106 cfu/mL |
| Bordetella hinzii | 1.40 x106 cfu/mL |
| Bordetella holmesii (ZeptoMetrix | 3.83 x106 cfu/mL |
| Bordetella holmesii (ATCC 51541) | 4.10 x106 cfu/mL |
| Bordetella holmesii (ATCC 700053) | 4.70 x106 cfu/mL |
| Bordetella holmesii (ATCC 700052) | 4.00 x106 cfu/mL |
| Bordetella parapertussis A747 | 1.00 x106 cfu/mL |
| Bordetella petrii | 6.26 x106 cfu/mL |
| Bordetella trematum | 9.24 x106 cfu/mL |
| Burkholderia cenocepacia | 2.35 x106 cfu/mL |
| Burkholderia cepacia | 2.52 x106 cfu/mL |
| Burkholderia multivorans | 1.95 x106 cfu/mL |
| Burkholderia thailandensis | 3.95 x106 cfu/mL |
| Chlamydia trachomatis | 7.83 x106 cfu/mL |
| Organism | Test Concentration |
| Chlamydophila pneumoniae | 2.10 x106 DNA copies/mL |
| Corynebacterium diptheriae | 4.00 x106 cfu/mL |
| Enterobacter aerogenes | 1.31 x106 cfu/mL |
| Enterococcus faecalis | 3.45 x106 cfu/mL |
| Escherichia coli | 8.42 x106 cfu/mL |
| Fusobacterium necrophorum | 3.10 x106 cfu/mL |
| Haemophilus influenzae | 2.13 x106 cfu/mL |
| Klebsiella pneumoniae | 1.61 x106 cfu/mL |
| Lactobacillus acidophilus | 2.00 x106 cfu/mL |
| Lactobacillus plantarum | 7.97 x106 cfu/mL |
| Legionella pneumophilia | 1.76 x106 cfu/mL |
| Moraxella catarrhalis | 9.90 x106 cfu/mL |
| Morganella morganii | 1.57 x106 cfu/mL |
| Mycobacterium avium | 1.84 x106 cfu/mL |
| Mycobacterium tuberculosis | 1.80 x106 cfu/mL |
| Mycoplasma pneumoniae | 3.16 x106 cfu/mL |
| Neisseria gonorrhoeae | 2.45 x106 cfu/mL |
| Neisseria meningitidis | 7.07 x106 cfu/mL |
| Neisseria mucosa | 1.66 x106 cfu/mL |
| Parvimonas micra | 1.55 x106 cfu/mL |
| Proteus mirabilis | 1.06 x106 cfu/mL |
| Proteus vulgaris | 3.40 x106 cfu/mL |
| Pseudomonas aeruginosa | 2.60 x106 cfu/mL |
| Staphylococcus aureus (MRSA) | 7.10 x106 cfu/mL |
| Staphylococcus epidermidis | 2.14 x106 cfu/mL |
| Stenotrophomonas maltophilia | 1.90 x106 cfu/mL |
| Streptococcus pneumoniae | 1.00 x106 cfu/mL |
| Streptococcus pyogenes | 1.29 x106 cfu/mL |
| Streptococcus salivarius | 1.70 x106 cfu/mL |
| Candida albicans | 3.00 x106 cfu/mL |
| Adenovirus 31 | 3.55 x105 TCID50/mL |
| Adenovirus 31 | 2.74 x107 DNA copies/mL |
| Coronavirus 229E | 1.51 x106 TCID50/mL |
| Coronavirus NL63 | 1.41 x105 TCID50/mL |
| Coronavirus OC43 | 8.51 x106 TCID50/mL |
| Coxsackievirus B4 | 1.08 x105 TCID50/mL |
| Coxsackievirus B5/10/2006 | 1.02 x105 TCID50/mL |
| Echovirus 6 | 1.02 x106 TCID50/mL |
| Echovirus 7 | 1.05 x105 TCID50/mL |
| Echovirus 9 | 1.41 x105 TCID50/mL |
17
18
Organism Test Concentration Echovirus 11 1.51 ×10° TCID50/mL 1.78 ×10° Enterovirus 70 TCID50/mL Enterovirus 71 4.17 ×10° TCID50/mL Epstein-Barr Virus 1.34 ×10° Virus particles/mL HSV Type 1 (Mclnytre) 6.65 ×10° TCID50/mL HSV Type 2 (G) 2.27 ×10° TCID50/mL 2.88 ×10° Influenza A/Mexico/4108/2009 Virus particles/mL Influenza B/Florida/04/2006 2.82 ×10° Virus particles/mL Measles virus 1.95 ×10° TCID50/mL Metapneumovirus A1 3.80 ×10° TCID50/mL 5.89 ×10° Mumps virus TCID50/mL 3.97 ×10° TCID50/mL Parainfluenza Type 1 (#2) Parainfluenza Type 2 (Greer) 3.15 ×10° TCID50/mL TCID50/mL Parainfluenza Type 3 (C234) 2.56 ×10° 1.37 ×106 Parainfluenza Type 4 (VR-1377) TCID50/mL Respiratory Syncytial Virus 1.15 ×106 TCID50/mL Rhinovirus 1A 1.26 ×10° TCID50/mL 1.70 x10° Varicella Zoster Virus DNA copies/mL
510(k) Summary
The Cross Reactivity study tested a panel of 79 microorganisms. This study determined that 1 of 4 Bordetella bronchiseptica strains (strain 4617) and 4 of 4 Bordetella holmesii strains tested were cross-reactive with the AmpliVue Bordetella Assay. These results can be expected as 5% of all Bordetella bronchiseptica strains and all Bordetella holmesii strains are known to carry the IS481 target sequence. These cross-reactive results are noted in the intended use and limitation sections.
Interference:
A study was conducted to determine if the AmpliVue "Bordetella assay is inhibited in the presence of a panel of seventeen (17) substances potentially present in specimens collected to test for Bordetella pertussis infection. Each of the potential interfering substances was tested in three replicates in the presence and absence of near LOD (2x) levels of B. pertussis in the AmpliVue Bordetella Assay. Substances were introduced into the assay at concentrations which were medically relevant.
| Common Name | Test Concentration |
|---|---|
| Cepacol Sore Throat Lozenges | 5% w/v |
| Halls Cherry Menthol-Lyptus Cough Drops | 15% w/v |
| Children's Dimetapp | 15% v/v |
| Chloraseptic Sore Throat Lozenges | 10% w/v |
19
510(k) Summary
| Ricola Original Swiss Sugar-Free Herb Cough Suppressant Throat Drops | 15% w/v |
|---|---|
| Sucrets Complete Lozenges - Vapor Cherry | 5% w/v |
| Mucin (Bovine Submaxillary Gland, Type I-S) | 5 mg/ml |
| Blood (human), EDTA anticoagulated | 5% v/v |
| Neo-Synephrine | 15% v/v |
| Afrin Nasal Spray Original | 15% v/v |
| Zicam Non-Drowsy Allergy Relief Nasal Gel | 5% v/v |
| Rite Aid Brand Saline Nasal Spray | 15% v/v |
| Zanamivir (Relenza) | 5 mg/ml |
| Tobramycin | 4 µg/ml |
| Mupirocin | 10 mg/ml |
| Oseltamivir Phosphate (Tamiflu) | 10 mg/ml |
| Ricola Original Swiss Sugar Free Herb Cough Suppressant Throat Drops | 15% w/v |
There was no evidence of interference caused by the substances tested.
Analytical Reactivity (Inclusivity):
The reactivity of the AmpliVue " Bordetella assay was evaluated against an additional six (6) strains of Bordetella pertussis at concentrations near the level of detection (LoD) of the assay.
Each strain was tested as three replicates in the AmpliVue "Bordetella. The study was performed in multiple experiments of no more than 14 assays per experiment. For each experiment, three replicates of up to four strains were performed, along with a positive and a negative run control. All seven strains were detected by the AmpliVue `Bordetella Assay.
| Bacterial Strain | Concentration
CFU/mL | Strain Detected
(Yes/No) |
|------------------|-------------------------|-----------------------------|
| 9797 | 2,358 | Yes |
| 9340 | 2,358 | Yes |
| BAA-1335 | 2,358 | Yes |
| BAA-589 | 2,358 | Yes |
| 51445 | 2,358 | Yes |
| 10380 | 2,358 | Yes |
- f. Assay cut-off:
20
Not applicable.
-
- Comparison studies:
- a. Method comparison with predicate device:
Not applicable
- b. Matrix comparison:
The compatibility of eight different types of media with the AmpliVue Bordetella assay was determined using Validation Lot reagents and the proposed workflow. Four strains of pre-titered Bordetella pertussis stocks were tested at 1x LoD in the presence of eight different types of media (Tris EDTA, Molecular Grade Water, Eswab, M4, M4-RT, M5, 0.85% Saline).
For Bordetella pertussis, 4 of 4 strains (A639, E431, BAA 1335, and 51445) were determined to be compatible with the 8 different media types when tested at the assay LoD.
3. Clinical studies:
- Clinical Sensitivity: a.
Performance characteristics of the AmpliVue® Bordetella Assay was established in the Spring to Summer of 2014 (April to August 2014) at four locations in the United States. Eight hundred forty two (842) fresh nasopharyngeal swab specimens were obtained from female and male patients suspected of having respiratory tract infection attributable to Bordetella pertussis from were collected and transported to each laboratory for testing with the AmpliVue® Bordetella Assay.
Clinical performance was based on comparison of the AmpliVue Bordetella Assay results to those obtained by Composite Reference Method that included two manufacturer validated, IS481-targeted real-time PCR assays (PCR1 and PCR2) followed by bi-directional sequencing from PCR positive specimens. The PCR1 and PCR2 assay protocols included 37 amplification cycles. Bi-directional sequencing was performed for all specimens producing amplicon prior to the end of 37-cycle amplification. Specimens were considered positive when bi-directional sequence sequencing results from either comparator PCR assay confirmed the presence of
21
Bordetella pertussis amplicon. Specimens were considered negative when neither comparator PCR assay produced Bordetella pertussis amplicon at the end of the 37cycles.
Eight hundred forty two (842) fresh nasopharyngeal swab specimens were tested as described above. Six (6) specimens (0.7%) were invalid (in both the initial and repeat test neither the T2 or control lines were detected) and have been removed from additional analysis. The table below details the comparison data of the AmpliVue Bordetella Assay and the Composite Reference Method for the remaining eight hundred thirty six (836) specimens.
| Composite Reference Method versus AmpliVue Bordetella Assay | ||||
|---|---|---|---|---|
| Composite Reference Method | ||||
| AmpliVue® Bordetella Assay | Positive | Negative | Total | |
| Positive | 64 | 15 | 79 | |
| Negative | 2 | 755 | 757 | |
| Total | ୧୧ | 770 | 836 | |
| 95% Cl | ||||
| Positive Percent Agreement | 64/66 | 97.0% | 89.6% to 99.2% | |
| Negative Percent Agreement | 755/770 | 98.1% | 96.8% to 98.8% |
-
b. Clinical specificity:
See Section 3a. -
Other clinical supportive data (when a. and b. are not applicable): ﻥ
Not applicable -
- Clinical cut-off:
Not applicable
- Clinical cut-off:
-
- Expected values:
The prevalence of Bordetella pertussis detected with the AmpliVue® Bordetella Assay has been calculated for the combined sites based on the age of the patient. Six (6)
- Expected values:
22
specimens (0.7%) were invalid (in both the initial and repeat test neither the T2 or control lines were detected) and have been removed from the Expected Values table. The table below presents the data for the remaining eight hundred forty two (842) specimens.
| Combined Study – Expected Values (N=836) | |||
|---|---|---|---|
| Bordetella pertussis | |||
| Age | Total # | Total | Prevalence |
| Positive | |||
| 22 years | 280** | 14 | 5.0% |
- Four (4) specimens were invalid
** Two (2) specimens were invalid
N. Other Supportive Instrument Performance Characteristics Data Not Covered In The "Performance Characteristics" Section above:
Not applicable.
P. Proposed Labeling:
The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10, 21 CFR 801.109, and the special controls.
Q. Conclusion:
The AmpliVue® Bordetella Assay is as safe and effective as the predicate device for the qualitative detection of Bordetella pertussis nucleic acids isolated from nasopharyngeal swab specimens obtained from patients suspected of having respiratory tract infection attributable to Bordetella pertussis.