The ProAdeno™+ Assay is a multiplex Real Time PCR in vitro diagnostic test for the qualitative detection of human Adenovirus (HAdV) DNA isolated and purified from nasopharyngeal (NP) swab specimens obtained from individuals exhibiting signs and symptoms of acute respiratory infection. This test is intended for use to aid in the diagnosis of HAdV infections in humans in conjunction with other clinical and laboratory findings. The test detects, but does not differentiate, serotypes 1-51.

Negative results do not preclude HAdV infection and should not be used as the sole basis for treatment or other patient management decisions.

Device Description

The ProAdeno+ Assay enables detection of human adenovirus and an Universal Internal Control.

An overview of the procedure is as follows:

Collect nasopharyngeal swab specimens from symptomatic patients using a polyester, rayon or nylon tipped swab and place into viral transport medium.
Add Universal Internal Control (UIC) to every sample to monitor for inhibitors present in the specimens.
Perform isolation and purification of nucleic acids using a MagNA Pure LC System (Roche) and the MagNA Pure Total Nucleic Acid Isolation Kit (Roche) or a NucliSENS® easyMAG™ System (bioMérieux) and the Automated Magnetic Extraction Reagents (bioMérieux).
Add purified nucleic acids to ProAdeno+ Supermix included in the ProAdeno+ Assay Kit. The ProAdeno+ Mix contains oligonucleotide primers, target-specific oligonucleotide probes, and a Taq DNA polymerase. The primers are complementary to a highly conserved region of human adenovirus. The probes are dual-labeled with a reporter dye attached to the 5'-end and a quencher dye attached to the 3'-end (see table below).
Perform amplification of DNA in a Cepheid SmartCycler® II instrument. In this process, the probe anneals specifically to the template followed by primer extension and amplification. The ProAdeno+ Assay is based on Tagman reagent chemistry, which utilizes the 5' - 3' exonuclease activity of the Taq polymerase to cleave the probe thus separating the reporter dye from the quencher. This generates an increase in fluorescent signal upon excitation from a light source. With each cycle, additional reporter dye molecules are cleaved from their respective probes, further increasing fluorescent signal. The amount of fluorescence at any given cycle is dependent on the amount of amplification products present at that time. Fluorescent intensity is monitored during each PCR cycle by the real-time instrument.

AI/ML Overview

The provided text describes the clinical performance and reproducibility testing for the ProAdeno™+ Assay. Here's a breakdown of the requested information:

Acceptance Criteria and Reported Device Performance

The document does not explicitly state pre-defined acceptance criteria values for sensitivity and specificity. However, the reported performance is presented in a table. For reproducibility, the overall percent agreement is stated.

Metric	Acceptance Criteria (Not explicitly stated in document)	Reported Device Performance
Clinical Performance
Sensitivity	Not explicitly stated	97.5% (95% CI: 87.1% - 94.3%)
Specificity	Not explicitly stated	95.6% (95% CI: 94.3% - 96.7%)
Reproducibility
Overall % Agreement	Not explicitly stated	99.2%

Note: The 95% CI for Sensitivity and Specificity appear to have a typo in the provided text (87.1% - 94.3% combined for both). Assuming the first value is for sensitivity and the second for specificity, the corrected representation would be:

Sensitivity: 97.5% (95% CI: 87.1% - 94.3%)
Specificity: 95.6% (95% CI: 94.3% - 96.7%)

However, the CI values are given as a range, which makes this interpretation potentially problematic. Without further clarification from the original document, the exact intended 95% CI for sensitivity and specificity remains ambiguous as presented. For the purpose of this analysis, I will retain the values as they appear in the source text, acknowledging the potential discrepancy in the CI range.

Study Details

2. Sample size used for the test set and the data provenance

Sample Size: 1167 nasopharyngeal (NP) swab samples were initially tested. One sample was excluded due to unresolved results, leading to a final test set of 1166 samples for the clinical performance analysis.
Data Provenance: The study was a prospective study conducted at 4 U.S. clinical laboratories from October 2009 to August 2010. Samples were NP swab specimens collected for routine respiratory viral testing by each site.

3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts

The document does not specify the number or qualifications of experts used for establishing the ground truth. It states that the reference method was "rapid culture (shell vial) followed by direct fluorescent antibody (DFA) screening and identification." Discrepant analysis for samples in disagreement was performed using "PCR primers obtained from literature followed by sequencing." This implies laboratory-based methods for ground truth, rather than expert human interpretation of images or other subjective data.

4. Adjudication method for the test set

The document describes a form of discrepant analysis. For samples where the ProAdeno+ Assay and the initial reference method (culture/DFA) disagreed, further testing was performed using PCR primers obtained from literature followed by sequencing. This served as the adjudicating method to establish the true status of discrepant samples.

5. If a multi reader multi case (MRMC) comparative effectiveness study was done

No, a multi-reader multi-case (MRMC) comparative effectiveness study was not done. The study evaluates the performance of a diagnostic assay (ProAdeno+ Assay) against a reference laboratory method, not human readers with or without AI assistance.

6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done

Yes, this was a standalone performance study. The ProAdeno+ Assay is an in vitro diagnostic test, and its performance was evaluated directly against a reference method without human interpretation as part of the primary outcome.

7. The type of ground truth used

The ground truth was established using a combination of laboratory methods:

Primary reference method: Rapid culture (shell vial) followed by direct fluorescent antibody (DFA) screening and identification.
Discrepant analysis ground truth: PCR primers obtained from literature followed by sequencing.

8. The sample size for the training set

The document does not specify a separate training set. The descriptions provided are for the clinical performance evaluation and reproducibility assessment. This is typical for a diagnostic test validation, where the focus is on evaluating the device's performance characteristics in a clinical setting rather than training a machine learning model.

9. How the ground truth for the training set was established

As no separate training set is mentioned or described, the method for establishing its ground truth is not applicable from the provided text.

Summary

{0}------------------------------------------------

K102952

Page 1 of 5 Date: December 3, 2010

510(k) SUMMARY

CONTACT

Emily Ziegler Gen-Probe Prodesse, Inc. W229 N1870 Westwood Dr. Waukesha. WI 53186

DEL - : 2010

NAME OF DEVICE

Trade Name: Regulation Number: Classification Name: · ProAdeno™+ Assay 21 CFR 866.3980 Respiratory viral panel multiplex nucleic acid assay

PREDICATE DEVICE

K063765, K081483, K091667 ID Tag Respiratory Virus Panel, Luminex Molecular . Diagnostics

INTENDED USE

Negative results do not preclude HAdV infection and should not be used as the sole basis for treatment or other patient management decisions.

{1}------------------------------------------------

PRODUCT DESCRIPTION .

The ProAdeno+ Assay enables detection of human adenovirus and an Universal Internal Control.

An overview of the procedure is as follows:

1. Collect nasopharyngeal swab specimens from symptomatic patients using a polyester, rayon or nylon tipped swab and place into viral transport medium.
1. Add Universal Internal Control (UIC) to every sample to monitor for inhibitors present in the specimens.
1. Perform isolation and purification of nucleic acids using a MagNA Pure LC System (Roche) and the MagNA Pure Total Nucleic Acid Isolation Kit (Roche) or a NucliSENS® easyMAG™ System (bioMérieux) and the Automated Magnetic Extraction Reagents (bioMérieux).
1. Add purified nucleic acids to ProAdeno+ Supermix included in the ProAdeno+ Assay Kit. The ProAdeno+ Mix contains oligonucleotide primers, target-specific oligonucleotide probes, and a Taq DNA polymerase. The primers are complementary to a highly conserved region of human adenovirus. The probes are dual-labeled with a reporter dye attached to the 5'-end and a quencher dye attached to the 3'-end (see table below).
1. Perform amplification of DNA in a Cepheid SmartCycler® II instrument. In this process, the probe anneals specifically to the template followed by primer extension and amplification. The ProAdeno+ Assay is based on Tagman reagent chemistry, which utilizes the 5' - 3' exonuclease activity of the Taq polymerase to cleave the probe thus separating the reporter dye from the quencher. This generates an increase in fluorescent signal upon excitation from a light source. With each cycle, additional reporter dye molecules are cleaved from their respective probes, further increasing fluorescent signal. The amount of fluorescence at any given cycle is dependent on the amount of amplification products present at that time. Fluorescent intensity is monitored during each PCR cycle by the real-time instrument.

Analyte	Gene Targeted	Probe Fluorophore	Absorbance Peak	Emission Peak	Instrument Channel
Adenovirus	hexon	FAM	495 nm	520 nm	FAM
Universal Internal Control	NA	Quasar 670	647 nm	667 nm	Cy5

{2}------------------------------------------------

SUBSTANTIAL EQUIVALENCE

Clinical Performance

Performance characteristics of the ProAdeno+ Assay were established during a prospective study at 4 U.S. clinical laboratories from October 2009- August 2010. Samples used for this study were nasopharyngeal (NP) swab specimens that were collected for routine respiratory viral testing by each site.

The reference method was rapid culture (shell vial) followed by direct fluorescent antibody (DFA) screening and identification.

A total of 1167 NP swab samples were tested with the ProAdeno+ Assay and by culture. One sample that initially gave unresolved results remained unresolved upon retesting with the ProAdeno+ Assay and is not included in the analysis below. The sample was culture negative.

Discrepant analysis for samples where ProAdeno+ Assay and culture results were in disagreement was performed using PCR primers obtained from literature followed by sequencing.

Results from Prospective Study

Adenovirus Comparison Results

	Reference Method
	Positive	Negative	Total	Comments
ProAdeno+ AssayPositive	39	49a	88	Sensitivity 97.5% (87.1% - 94.3%)95% CI
ProAdeno+ AssayNegative	1b	1077	1078	Specificity 95.6% (94.3% - 96.7%)95% CI
Total	40	1126	1166

435 samples positive for HAdV by bi-directional sequence analysis, 14 samples negative for HAdV by bi-directional sequence analysis.

b1 sample negative for HAdV by bi-directional sequence analysis

{3}------------------------------------------------

Age Group	Total (N)	ProspectiveTotal # Positive byProAdeno+ Assay	Observed Prevalence
< 2 years	485	58	12.0%
2-5 years	184	18	9.8%
6-11 years	101	7	6.9%
12-18 years	67	3	4.5%
19-64 years	240	2	0.8%
>65 years	89	0	0%
Total	1166	88	7.5%

{4}------------------------------------------------

Reproducibility

The reproducibility of the ProAdeno+ Assay was evaluated at 3 laboratory sites. Reproducibility was assessed using a panel of 12 simulated clinical samples that included two adenovirus serotypes at medium and low positive levels (near the assay limit of detection, > 95% positive) and two high negative samples (high negative-1 at 0.1xLoD; high negative-2 at 0.001xLoD). Panels and controls were tested at each site by 2 operators for 5 days (12 samples and 3 controls X 2 operators X 5 days X 3 sites = 450). The overall percent agreement for the ProAdeno+ Assay was 99.2%.

	PanelMember IDConcentration	HAdV-3 highnegative-2"0.001X LoD	HAdV-3 highnegative-10.1XLoD	HAdV-3 low positive2XLoD	HAdV-3 mediumpositive10XLoD	HAdV-31 highnegative-2"0.001X LoD	HAdV-31 highnegative-10.1XLoD	HAdV-31 lowpositive2XLoD	HAdV-31 mediumpositive10XLoD	ExtractionControlN/A	Adenovirus DNAControlN/A	Negative Control"N/A	Total % Agreement
Site 1°	Agreementwith ExpectedResult	15/15	5/15b	15/15	15/15	15/15	10/15b	15/15	15/15	10/10	10/10	10/10	120/120d (100%)
	Mean CTValue	36.9	40.4c	35.8	33.9	37.2	38.6c	33.8	29.3	31.8	32.5	36.7
	% CV	2.5	2.8c	1.2	1.0	2.2	6.2c	1.7	1.4	0.5	0.8	1.0
Site 2r	Agreementwith ExpectedResult	14/14	3/16b	16/16	14/14	15/16	6/14b	14/14	16/16	11/11	11/11	11/11	122/123d (99.2%)
	Mean CTValue	36.5	39.8c	36.9	34.9	36.9	39.6c	35.1	30.9	34.9	33.3	36.5
	% CV	1.0	3.5c	1.3	0.9	2.0	6.6c	2.8	3.4	2.2	1.2	1.1
Site 3r	Agreementwith ExpectedResult	15/15	2/15b	15/15	15/15	13/15	3/15b	15/15	15/15	10/10	10/10	10/10	118/120d (98.3%)
	Mean CTValue	36.5	40.1c	36.7	34.6	36.3	39.2c	34.9	30.8	32.3	32.5	36.5
	% CV	1.1	5.3c	2.6	1.1	1.3	6.1c	1.3	1.1	1.0	1.5	1.1
	TotalAgreementwith ExpectedResult	44/44	10/46b	46/46	44/44	43/46	19/44b	44/44	46/46	31/31	31/31	31/31	360/363d (99.2%)
	95% CI	92.0-100%	N/A	92.3-100	92.0-100%	82.1-98.6%	N/A	92.0-100%	92.3-100	88.8-100%	88.8-100%	88.8-100%	98.0-99.9%
	Overall MeanCT Value	36.6	40.2c	36.5	34.4	36.8	39.0c	34.6	30.3	33.1	32.8	36.6
	Overall% CV	1.8	3.1c	2.2	1.6	2.1	6.1c	2.6	3.3	4.5	1.7	1.1

4Mean Cr calculated from Universal Internal Control

'Number of positive samples

· Average and %CV based on number of positive samples

4Does not include intermediate samples as those are at a concentration that is not reproducible

*Performed study using the bioMérieux NucliSENS easyMAG

Performed study using the Roche MagNA Pure

{5}------------------------------------------------

Image /page/5/Picture/0 description: The image shows the logo for the Department of Health & Human Services USA. The logo features a stylized eagle with three stripes representing the department's mission. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES USA" is arranged in a circular pattern around the eagle.

Food and Drug Administration 10903 New Hampshire Avenue Document Mail Center - WO66-0609 Silver Spring, MD 20993-0002

Gen-Probe Prodesse, Inc. c/o Emily Ziegler Research Associate III W229 N1870 Westwood Dr. Waukesha. WI 53186

DEC - 3 2010

Re: K102952

Trade/Device Name:	ProAdeno™+
Regulation Number:	21 CFR §866.3980
Regulation Name:	Respiratory viral panel multiplex nucleic acid assay
Regulatory Class:	Class II
Product Code:	OCC
Dated:	October 4, 2010
Received:	October 5, 2010

Dear Ms. Ziegler:

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 In Incolumn include 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 1 (1) The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbrandi, insand adulteration.

ff your device is classified (see above) into class II (Special Controls), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21. Code of Federal Regulations (CFR), Parts 800 to 895. 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); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820). This letter will allow you to begin marketing your device as described in your Section 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed

{6}------------------------------------------------

Page 2 - Emily Ziegler

predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific advice for your device on our labeling regulation (21 CFR Parts 801 and 809), please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 796-5450. 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/McdicalDevices/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 Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely yours,

Satiyagn

Sally A. Hojvat, M.Sc., P Director Division of Microbiology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

{7}------------------------------------------------

Indications for Use

510(k) Number (if known): K102952

Device Name: ProAdeno™+ Assay

Indication for Use:

Negative results do not preclude HAdV infection and should not be used as the sole basis for treatment or other patient management decisions.

Prescription Use X (21 CFR Part 801 Subpart D) And/Or

Over the Counter Use (21 CFR Part 801 Subpart C)

(PLEASE DO NOT WRITE BELOW THIS LINE; CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of In Vitro Diagnostic Device Evaluation and Safety (OIVD)

Uve Schuf

Division Sign-Off Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) lo2952

Regulation Number and Section

§ 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.