(263 days)
No
The description focuses on standard nucleic acid amplification and detection technologies (NAAT, TMA, HPA) and does not mention any AI or ML components for data analysis or interpretation.
No
The device is an in vitro diagnostic test for detecting Neisseria gonorrhoeae, aiming to aid in diagnosis, not to treat or alleviate a disease.
Yes
The "Intended Use / Indications for Use" section explicitly states that the assay is "to aid in the diagnosis of gonococcal urogenital disease."
No
The device description clearly details a complex in vitro diagnostic (IVD) assay involving reagents, target capture, amplification, and detection using a physical system (Panther System). This is a hardware-based medical device with associated software for control and analysis, not a software-only device.
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 Aptima Neisseria gonorrhoeae (GC) Assay is an "in vitro qualitative nucleic acid amplification (NAAT) for the detection of ribosomal RNA (rRNA) from Neisseria gonorrhoeae (GC) to aid in the diagnosis of gonococcal urogenital disease". The phrase "in vitro" is a key indicator of an IVD, meaning it is used outside of the body.
- Device Description: The description details a laboratory test that analyzes biological specimens (urine) using chemical and molecular methods (nucleic acid amplification, hybridization protection assay) to detect a specific target (GC rRNA). This is characteristic of an IVD.
- Clinical Study: The document describes a clinical study where the device was used to test patient specimens in a laboratory setting to evaluate its performance in aiding diagnosis. This is a standard process for validating IVDs.
- Predicate Device: The mention of a "Predicate Device(s)" with a K number (K063664) indicates that this device is being compared to a previously cleared device, which is a regulatory pathway for IVDs.
All of these points strongly support the classification of this device as an In Vitro Diagnostic.
N/A
Intended Use / Indications for Use
The Aptima Neisseria gonorrhoeae (GC) Assav is an in vitro qualitative nucleic acid amplification (NAAT) the detection of ribosomal RNA (rRNA) from Neisseria gonorrhoeae (GC) to aid in the diagnosis of gonocccal urogenital disease using the Panther System. The assay may be used to test male urine specimens from symptomatic and asymptomatic individuals.
Product codes (comma separated list FDA assigned to the subject device)
LSL, QEP
Device Description
Clearance of this pre-market application will allow the Aptima Neisseria gonorrhoeae Assay to be performed on the Panther system using male urine specimens.
The Aptima GC assay is a target amplification nucleic acid probe test for in vitro qualitative detection of ribosomal RNA (rRNA) from Neisseria gonorrhoeae (GC). The Aptima Neisseria gonorrhoeae Assay combines the technologies of target capture, transcription-mediated amplification (TMA), and hybridization protection assay (HPA).
Specimens are collected and transferred into their respective specimen transport tubes. The transport solution in these tubes releases the rRNA target and protects it from degradation during storage. When the Aptima Neisseria gonorrhoeae Assay is performed in the laboratory, the target rRNA molecule is isolated from the specimens by use of a capture oligomer via target capture that utilizes magnetic microparticles. The capture oligomer contains a sequence complementary to a specific region of the target molecule as well as a string of deoxyadenosine residues. During the hybridization step, the sequence specific region of the capture oligomer binds to a specific region of the target molecule. The capture oligomer:target complex is then captured out of solution by decreasing the temperature of the reaction to room temperature. This temperature reduction allows hybridization to occur between the deoxyadenosine region on the capture oligomer and the poly-deoxythymidine molecules that are covalently attached to the magnetic particles. The micro particles, including the captured target molecule bound to them, are pulled to the side of the reaction vessel using magnets and the supernatant is aspirated. The particles are washed to remove residual specimen matrix that may contain amplification reaction inhibitors. After the target capture steps are completed, the specimens are ready for amplification.
Target amplification assays are based on the ability of complementary oligonucleotide primers to specifically anneal and allow enzymatic amplification of the target nucleic acid strands. The Hologic TMA reaction replicates a specific region of the 16S rRNA from GC via DNA intermediates. A unique set of primers is used for the target molecule. Detection of the rRNA amplification product sequences (amplicon) is achieved using nucleic acid hybridization. A single-stranded chemiluminescent DNA probe, which is complementary to a region of the target amplicon, is labeled with an acridinium ester molecule. The labeled DNA probe combines with amplicon to form stable RNA:DNA hybrids. The Selection Reagent differentiates hybridized from unhybridized probe, eliminating the generation of signal from unhybridized probe. During the detection step, light emitted from the labeled RNA:DNA hybrids is measured as photon signals in a luminometer, and are reported as Relative Light Units (RLU).
The device reagents are identical to the Aptima Neisseria gonorrhoeae Assay reagents for use on the Tigris DTS system but are intended for use on the Panther system with different specimen type indications. The Panther and Tigris DTS systems use the same principles of operation.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Urogenital
Indicated Patient Age Range
Not Found
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
Not Found
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Analytical Studies:
- Analytical Sensitivity / Limit of Detection Study: LoD on the Panther was below 125 CFU/mL (250 fg GC rRNA/assay). For Neisseria gonorrhoeae ATCC 49226, LoD was 0.04933 CFU/mL. For stain X/NCTC 13820, LoD was 0.03986 CFU/mL.
- Within-lab Precision Study: N=162 replicates for each panel member. Agreement to expected results was 100% for all four panel members (Low Positive, Moderate Positive, High Positive, Negative).
- Analytical Specificity: Evaluated 155 culture isolates (87 urogenital, 68 phylogenetic cross-section). All organisms except C. psittaci, C. pneumoniae, U. urealyticum, C. trachomatis and viruses were tested at 1.0 x 10^6 cells/assay.
- Interfering Substances: No interference observed with tested substances (10% blood, contraceptive jelly, spermicide, moisturizer, hemorrhoidal anesthetic, body oil, powder, antifungal cream, vaginal lubricants, feminine spray, leukocytes, 30% blood, urine analytes, protein, glucose, ketones, bilirubin, nitrate, urobilinogen, pH 4, pH 9, cellular debris, vitamins, minerals, acetaminophen, aspirin, ibuprofen, mucin, seminal fluid). No inhibitors of amplification observed.
- Carryover Study: Multi-run analytical study on three Panther Systems. Total of 2941 negative samples. Overall carryover rate was 0.07% with a 95% confidence interval of 0.02–0.25%.
- Run-Size Validity: Negative and Positive panel member results produced 100% agreement with expected results, with no difference in performance between the front and back of the runs.
- Control Validity: Acceptance criteria met. Results at time-point 0 hour and time-point 30 hour yielded expected results and control RLUs were within expected range. Controls properly manage run validity over 24-hours.
- Control Effectiveness: Performance of GC Controls correctly predicted sample results in 8 out of 8 conditions tested. Results met acceptance criteria.
- Environmental Conditions: Negative and Positive panel member results produced 100% agreement with expected results at specified environmental limits (15-30°C and 20-85% relative humidity).
- Reproducibility Study: Conducted at two external US laboratories and Hologic. Testing with two reagent lots and six operators. N=108 for Negative, N=105 for Low Positive, N=107 for Positive. Agreement with expected results was 100% for all panel members.
Clinical Study:
- Study Type: Prospective, multi-center clinical study.
- Sample Size: 2085 symptomatic and asymptomatic men enrolled. 1958 evaluable male urine specimens included in the analyses.
- Key Results: Calculated sensitivity is 98.4% (95% CI: 94.4%-99.6%) for male urine. Overall specificity estimate is 99.9% (95% CI: 99.7%-100%) for male urine. The device performs as intended and performs as safe and as effective compared to the predicate device.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Sensitivity: 98.4% (95% CI: 94.4%-99.6%) for male urine.
Specificity: 99.9% (95% CI: 99.7%-100%) for male urine.
Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.
Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.
Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).
Not Found
§ 866.3390
Neisseria spp. direct serological test reagents.(a)
Identification. Neisseria spp. direct serological test reagents are devices that consist of antigens and antisera used in serological tests to identifyNeisseria spp. from cultured isolates. Additionally, some of these reagents consist ofNeisseria spp. antisera conjugated with a fluorescent dye (immunofluorescent reagents) which may be used to detect the presence ofNeisseria spp. directly from clinical specimens. The identification aids in the diagnosis of disease caused by bacteria belonging to the genusNeisseria, such as epidemic cerebrospinal meningitis, meningococcal disease, and gonorrhea, and also provides epidemiological information on diseases caused by these microorganisms. The device does not include products for the detection of gonorrhea in humans by indirect methods, such as detection of antibodies or of oxidase produced by gonococcal organisms.(b)
Classification. Class II (performance standards).
0
Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food & Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.
January 26, 2024
Hologic, Inc. Jon Kukowski Regulatory Affairs Specialist 10210 Genetic Center Drive San Diego, California 92121
Re: K231329
Trade/Device Name: Aptima Neisseria gonorrhoeae Assay Regulation Number: 21 CFR 866.3390 Regulation Name: Neisseria Spp. Direct Serological Test Reagents Regulatory Class: Class II Product Code: LSL, QEP Dated: December 28, 2023 Received: December 28, 2023
Dear Jon Kukowski:
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 (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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database available at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. 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.
1
Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).
Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30. Design controls; 21 CFR 820.90. Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review, the QS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).
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 of medical device-related adverse events) (21 CFR Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
2
Sincerely,
Himani Bisht -S
Himani Bisht, Ph.D. Assistant Director Viral Respiratory and HPV Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health
Enclosure
3
Indications for Use
510(k) Number (if known) K231329
Device Name Aptima Neisseria gonorrhoeae Assay
Indications for Use (Describe)
The Aptima Neisseria gonorrhoeae (GC) Assav is an in vitro qualitative nucleic acid amplification (NAAT) the detection of ribosomal RNA (rRNA) from Neisseria gonorrhoeae (GC) to aid in the diagnosis of gonocccal urogenital disease using the Panther System. The assay may be used to test male urine specimens from symptomatic and asymptomatic individuals.
| Type of Use (Select one or both, as applicable) | |
|---|---|
| Prescription Use (Part 21 CFR 801 Subpart D) | Over-The-Counter Use (21 CFR 801 Subpart C) |
CONTINUE ON A SEPARATE PAGE IF NEEDED.
This section applies only to requirements of the Paperwork Reduction Act of 1995.
DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.
The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:
Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov
"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."
4
510(k) Summary
Contact Details
Applicant Name: Hologic, Inc. Applicant Address: 10210 Genetic Center Drive San Diego CA 92121 United States Applicant Contact Telephone: 858-410-8245 Applicant Contact: Jon Kukowski Applicant Contact Email: jonathan.kukowski@hologic.com
Device Name
Device Trade Name: Aptima Neisseria gonorrhoeae Assay Common Name: Neisseria spp. direct serological test reagents Classification Name: DNA-Reagents, Neisseria Regulation Number: 866.3390 Regulatory Class: Class II Product Code: LSL
Legally Marketed Predicate Devices
Predicate #: K063664 Predicate Trade Name: Aptima Neisseria gonorrhoeae Assay Product Code: LSL
Device Description Summary
Clearance of this pre-market application will allow the Aptima Neisseria gonorrhoeae Assay to be performed on the Panther system using male urine specimens.
The Aptima GC assay is a target amplification nucleic acid probe test for in vitro qualitative detection of ribosomal RNA (rRNA) from Neisseria gonorrhoeae (GC). The Aptima Neisseria gonorrhoeae Assay combines the technologies of target capture, transcription-mediated amplification (TMA), and hybridization protection assay (HPA).
Specimens are collected and transferred into their respective specimen transport tubes. The transport solution in these tubes releases the rRNA target and protects it from degradation during storage. When the Aptima Neisseria gonorrhoeae Assay is performed in the laboratory, the target rRNA molecule is isolated from the specimens by use of a capture oligomer via target capture
5
that utilizes magnetic microparticles. The capture oligomer contains a sequence complementary to a specific region of the target molecule as well as a string of deoxyadenosine residues. During the hybridization step, the sequence specific region of the capture oligomer binds to a specific region of the target molecule. The capture oligomer:target complex is then captured out of solution by decreasing the temperature of the reaction to room temperature. This temperature reduction allows hybridization to occur between the deoxyadenosine region on the capture oligomer and the poly-deoxythymidine molecules that are covalently attached to the magnetic particles. The micro particles, including the captured target molecule bound to them, are pulled to the side of the reaction vessel using magnets and the supernatant is aspirated. The particles are washed to remove residual specimen matrix that may contain amplification reaction inhibitors. After the target capture steps are completed, the specimens are ready for amplification.
Target amplification assays are based on the ability of complementary oligonucleotide primers to specifically anneal and allow enzymatic amplification of the target nucleic acid strands. The Hologic TMA reaction replicates a specific region of the 16S rRNA from GC via DNA intermediates. A unique set of primers is used for the target molecule. Detection of the rRNA amplification product sequences (amplicon) is achieved using nucleic acid hybridization. A single-stranded chemiluminescent DNA probe, which is complementary to a region of the target amplicon, is labeled with an acridinium ester molecule. The labeled DNA probe combines with amplicon to form stable RNA:DNA hybrids. The Selection Reagent differentiates hybridized from unhybridized probe, eliminating the generation of signal from unhybridized probe. During the detection step, light emitted from the labeled RNA:DNA hybrids is measured as photon signals in a luminometer, and are reported as Relative Light Units (RLU).
The device reagents are identical to the Aptima Neisseria gonorrhoeae Assay reagents for use on the Tigris DTS system but are intended for use on the Panther system with different specimen type indications. The Panther and Tigris DTS systems use the same principles of operation.
6
Intended Use
The Aptima Neisseria gonorrhoeae (GC) Assay is an in vitro qualitative nucleic acid amplification (NAAT) test for the detection of ribosomal RNA (rRNA) from Neisseria gonorrhoeae (GC) to aid in the diagnosis of gonococcal urogenital disease using the Panther System. The assay may be used to test urine from symptomatic and asymptomatic male individuals.
Indications for Use Comparison
This pre-market application is to clear the Aptima Neisseria gonorrhoeae Assay on an additional platform, the Panther system, for use with male urine specimen type indications. The analytical and clinical study results demonstrate that the Aptima Neisseria gonorrhoeae Assay on the Panther system performs comparably to the predicate device in detecting rRNA of GC from the designated specimen type and support a substantial equivalence decision.
Technological Comparison
The Aptima Neisseria gonorrhoeae Assay incorporates the technologies of target capture, Transcription-Mediated Amplification (TMA), and Hybridization Protection Assay (HPA), which are described in previous 510(k) submissions for the Aptima Neisseria gonorrhoeae Assay including K043144, cleared in March 2005. The Aptima Neisseria gonorrhoeae Assay and system technologies remain unchanged in this submission, including biochemical (e.g., in vitro nucleic acid probe test) and physical detection (e.g., chemiluminescence) technologies.
Non-Clinical and/or Clinical Tests Summary & Conclusions
The following studies were conducted to support performance of the Aptima Neisseria gonorrhoeae Assay on the Panther system using male urine specimens.
Analytical Studies
Analytical Sensitivity
Studies were conducted to verify the analytical sensitivity (concentration of analyte giving ≥95% positivity) of the Aptima Neisseria gonorrhoeae Assay on the Panther system by testing panels made by spiking GC organisms into male urine pools, Results showed that the LoD on the Panther was below 125 CFU/mL (250 fg GC rRNA/assay).
7
Limit of Detection Study
The limit of detection (LoD) was tested and confirmed with sensitivity panels prepared using two strains of GC organisms spiked into pooled negative urine. Testing evaluated one antibiotic susceptible strain; Neisseria gonorrhoeae ATCC 49226 (GP1803), and one antibiotic resistant stain; Neisseria gonorrhoeae WHO X/NCTC 13820 (GP2730). Sensitivity panels were tested on three Panther instruments with two reagent lots. At least 20 replicates were run for each concentration for each reagent lot for each strain. LoD, defined as the target concentration that can be detected in 95% of the replicates tested for urine specimens is 0.04933 CFU/mL for ATCC 49226 and 0.03986 CFU/mL for stain X/NCTC 13820.
Within-lab Precision Study
Precision was measured using positive panel members consisting of Neisseria gonorrhea cells at three different concentrations approximately: 3x LoD (Low Positive), >3x and 10x LoD (High positive). Cells were spiked into pooled negative urine matrix (Urine mixed 1:1 with Urine Transport Media). The negative panel member consisted of unspiked negative urine matrix. Testing was conducted over the course of at least 20 nonconsecutive days using two lots of reagents on three Panther systems by three operators performing at least two daily runs. Agreement to expected results was 100% for all four panel members. The precision of the signal for each panel is shown in table below.
| Panel
Memb
er | N.
gonorrhoeae
concentrat
ion
(CFU/mL) | N | Mean
kRL
U | Agreement to
Expected Result | Between-
Lot
SD | Between-
Lot
CV
(%) | Between-
Instrumen
t
SD | Between-
Instrumen
t
CV
(%) | Between-
Operator
SD | Between-
Operator
CV
(%) | Between-
Day
SD | Between-
Day
CV
(%) | Between-
Run
SD | Between-
Run
CV
(%) | Within-
Run
SD | Within-
Run
CV
(%) | Total
SD | Total
CV
(%) |
|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------|---------|------------------|---------------------------------|-----------------------|------------------------------|----------------------------------|-----------------------------------------|----------------------------|-----------------------------------|-----------------------|------------------------------|-----------------------|------------------------------|----------------------|-----------------------------|-------------|--------------------|
| Low
positive | 0.1431 | 16
2 | 520.
1 | 100% | 65.0
5 | 12.5
1 | 38.3
9 | 7.38 | 0.00 | 0.00 | 14.8
4 | 2.8
5 | 20.1
1 | 3.8
7 | 62.3
4 | 11.9
9 | 101.
07 | 19.4
3 |
| Moderate
positive | 0.2417 | 16
2 | 809.
4 | 100% | 90.3
2 | 11.1
6 | 52.7
4 | 6.52 | 7.71 | 0.95 | 15.0
0 | 1.8
5 | 24.0
9 | 2.9
8 | 84.3
6 | 10.4
2 | 137.
55 | 16.9
9 |
| High
positive | 0.5426 | 16
2 | 1529
.4 | 100% | 182.
18 | 11.9
1 | 78.4
1 | 5.13 | 19.0
9 | 1.25 | 51.4
5 | 3.3
6 | 0.00 | 0.0
0 | 111.
09 | 7.26 | 233.
86 | 15.2
9 |
| Negative | 0.000 | 16
2 | 4.2 | 100% | 1.19 | 28.3
6 | 0.87 | 20.8
0 | 0.51 | 12.2
8 | 0.00 | 0.0
0 | 0.00 | 0.0
0 | 3.52 | 84.1
7 | 3.85 | 92.0
4 |
| Note: The analysis was performed using SAS PROC MIXED, which applies a lower boundary of 0 to all variance components in the model by default. If a variance
component is 0, then SD=0 and CV=0%. | | | | | | | | | | | | | | | | | | |
8
Analytical Specificity
A total of 155 culture isolates were evaluated using the Aptima Neisseria gonorrhoeae Assay on the DTS system or Panther system. These isolates included 87 organisms that may be isolated from the urogenital tract and 68 additional organisms that represent a phylogenetic cross-section of organisms. The tested organisms included bacteria, fungi, yeast, parasites and viruses. All organisms except C. psittaci, C. pneumoniae, U. urealyticum, C. trachomatis and the viruses were tested at 1.0 x 106 cells/ assay. C. psittaci VR601 was tested at 8.0 x 104 cells/assay and C. psittaci VR125 was tested at 1.0 x 105 cells/assay. C. pneumoniae was tested at 4 x 103 cells/assay and U. urealyticum was tested at 6.7 x 106 cells/assay. C. trachomatis was tested at 1.0 x 104 IFU/mL. The viruses were tested as follows: (a) herpes simplex virus I: 2.5 x 104 TCID50/assay, (b) herpes simplex virus II: 6.0 x 104 TCID50/assay, (c) human papillomavirus 16: 2.9 x 106 DNA copies/assay and (d) cytomegalovirus: 4.8 x 105 cells/assay.
Interfering Substances
For a nucleic acid amplification assay, analytical specificity with respect to potentially interfering substances is largely determined by the chemistry of the assay (e.g. oligonucleotide sequences) rather than by the platform. Because the reagents for the Aptima Neisseria gonorrhoeae Assay are identical between DTS and Panther systems, data generated on the DTS system supports the performance of the assay on Panther system.
The following interfering substances were individually spiked into urine specimens: 10% blood, contraceptive jelly, spermicide, moisturizer, hemorrhoidal anesthetic, body oil, powder, antifungal cream, vaginal lubricants, feminine spray and leukocytes (1.0 x 10° cells/mL). The following interfering substances were individually spiked into urine specimens: 30% blood, urine analytes, protein, glucose, ketones, bilirubin, nitrate, urobilinogen, pH 4 (acidic), pH 9 (alkaline), leukocytes (1.0 x 106 cells/mL), cellular debris, vitamins, minerals, acetaminophen, aspirin and ibuprofen. All were tested for potential assay interference in the absence and presence of GC at the estimated rRNA equivalent of 50 GC cells/assay (250 fg/assay). The rRNA equivalents were calculated based on the genome size and estimated DNA:RNA ratio/cell of each organism. No interference was observed with any of the tested substances. No inhibitors of amplification were observed in the Aptima Neisseria gonorrhoeae Assay.
9
Mucin and seminal fluid were tested on the Panther system by spiking into pooled negative urine negative in the presence of Neisseria gonorrhoeae ATCC 49226. Mucin at (0.1% Vol/vol) and seminal fluid (at 1% vol/vol) did not interfere with the assay.
Carryover Study
A multi-run analytical study was conducted using spiked panels on three Panther Systems. Carryover was assessed using approximately 20% high titer GC samples (>2 x 10 cells/mL, or rRNA equivalent) dispersed between negative samples. Testing was carried out using 5 runs on each of three Panther Systems with a total of 2941 negative samples. The overall carryover rate was 0.07% with a 95% confidence interval of 0.02–0.25%.
Run-Size Validity
This study was to confirm that there are no front-to-back positional effects within a run for the Aptima Neisseria gonorrhoeae Assay on the Panther system. Panels and controls with known concentrations were tested and results were examined for front-to-back effects when compared to expected results. Negative and Positive panel member results produced 100% agreement with the expected results, with no difference in performance between the front and back of the runs.
Control Validity
This study was to demonstrate that Aptima Neisseria gonorrhoeae Assay run controls meet performance criteria and properly control for run validity over the timeframe allowed by the Panther software. The acceptance criteria for this study were met. Results at time-point 0 hour and time-point 30 hour (24 hours + 25%) yielded the expected results for the study and the control RLUs were within the expected range. Run controls met performance criteria and properly control for run validity over the 24-hours control validity timeframe allowed by the Panther software.
Control Effectiveness
This study was to demonstrate that the Aptima Neisseria gonorrhoeae Assay run controls are properly invalidated under fault conditions that are not detected by instrument process controls and that may exist during assay processing. The performance of the GC Controls correctly
10
predicted the sample results in 8 out of 8 of the conditions tested for the assay when run on the Panther system. Results met the acceptance criteria of the study and demonstrated that performance of the Aptima Neisseria gonorrhoeae Assay controls properly determine run and test validity under fault conditions that are not detected by Panther system process controls.
Environmental Conditions
This study was to demonstrate that the Aptima Neisseria gonorrhoeae Assay on the Panther system meets performance requirements at the limits of Panther system environmental conditions for external ambient temperature (15-30°C) and relative humidity (20-85%) specifications. Target panels at or below the sensitivity claims for the assays were tested. Negative and Positive panel member results produced 100% agreement with the expected results. The Aptima Neisseria gonorrhoeae Assay on the Panther system meets performance requirements at the limits of Panther system environmental conditions for external ambient temperature (15-30°C) and relative humidity (20-85%) specifications.
Reproducibility Study
Aptima Neisseria gonorrhoeae Assay reproducibility was evaluated on the Panther system at two external US laboratories and at Hologic. Testing was performed using two lots of assay reagents and a total of six operators (two at each site, testing was performed over at least six days.
Reproducibility panel members were created using clinical urine specimens in urine transport medium (UTM). The GC positive panel members were created by diluting GC positive clinical urine specimens with volume from pooled GC negative clinical urine specimens to achieve the appropriate targeted RLU results (low RLU positive or positive). The negative panel member was created using pooled GC negative clinical urine specimens.
The agreement with expected results was 100% for all panel members.
The table below shows the signal variability of assay RLU results for each panel member between sites, between operators, between lots, between runs, within runs, and overall (Total). Only samples with valid results were included in the analyses.
11
| Panel
Member | Target
RLU
(x1000) | N | Mean
RLU
(x1000) | Between
Sites | | Between
Operators | | Between
Lots | | Between
Runs | | Within
Runs | | Total | |
|-----------------|--------------------------|-----|------------------------|------------------|-------|----------------------|-------|-----------------|-------|-----------------|-------|----------------|-------|-------|-------|
| | | | | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) | SD | CV(%) |
| Negative |