(85 days)
Not Found
No
The device description and performance studies focus on a standard real-time PCR assay for nucleic acid detection. There is no mention of AI or ML in the text.
No
The Lyra™ Adenovirus Assay is an in vitro diagnostic test designed to detect viral DNA, aiding in diagnosis. It does not provide any form of therapy or treatment.
Yes
The product is described as an "in vitro diagnostic test" and its "intended use of this test is to aid in the diagnosis of HAdV".
No
The device description explicitly states it is a real-time PCR in vitro diagnostic test that utilizes an ABI 7500 Fast Dx Real-Time PCR Instrument and involves the detection of viral nucleic acids using primers and fluorescent-labeled probes. This indicates the device includes hardware components and reagents, not just software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use/Indications for Use: The description explicitly states it is an "in vitro diagnostic test" and its intended use is to "aid in the diagnosis of HAdV in conjunction with other clinical and laboratory findings." This clearly aligns with the definition of an IVD, which is used to examine specimens from the human body to provide information for diagnosis, treatment, or prevention of disease.
- Device Description: The description further reinforces this by detailing how the test works on "viral DNA isolated from a patient sample" and uses "target-specific primers and fluorescent-labeled probes that hybridize to conserved regions in the genomes of adenovirus and the PRC." This describes a laboratory test performed outside the body on a biological specimen.
- Performance Studies: The inclusion of performance studies (prospective and retrospective) with metrics like sensitivity, specificity, positive percent agreement, and negative percent agreement are standard for demonstrating the clinical performance of an IVD.
- Predicate Device: The mention of a predicate device (K121942; Adenovirus R-gene® US) which is also an IVD, further confirms the regulatory classification of this device.
Therefore, based on the provided information, the Lyra™ Adenovirus Assay is definitively an In Vitro Diagnostic device.
N/A
Intended Use / Indications for Use
The Lyra™ Adenovirus Assay is a real-time polymerase chain reaction (PCR) in vitro diagnostic test for the qualitative detection of human adenovirus (HAdV) viral DNA isolated from nasal and nasopharyngeal swab specimens obtained from individuals exhibiting signs and symptoms of acute respiratory infections. The intended use of this test is to aid in the diagnosis of HAdV in conjunction with other clinical and laboratory findings.
The test detects, but does not differentiate. HAdV species (A, B, C, D, E, and F) or serotypes (HAdV 1-52).
Negative results do not preclude HAdV infection and should not be used as the sole basis for diagnosis, treatment or other patient management decisions.
Product codes (comma separated list FDA assigned to the subject device)
OCC
Device Description
The Lyra™ Adenovirus Assay is a real-time polymerase chain reaction (PCR) in vitro diagnostic test for the qualitative detection of human adenovirus (HAdV) viral DNA. The assay detects viral nucleic acids that have been extracted from a patient sample. A real-time PCR reaction is carried out using the ABI 7500 Fast Dx Real-Time PCR Instrument under optimized conditions in a single tube, generating amplicons for adenovirus and the Process Control (PRC). Identification of human adenovirus (HAdV) and the PRC occurs by the use of target-specific primers and fluorescent-labeled probes that hybridize to conserved regions in the genomes of adenovirus and the PRC. The HAdV target is labeled with FAM dye, and the PRC target is labeled with Cy5 dye.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
nasal and nasopharyngeal swab specimens
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
Prospective Study:
- Sample Size: 1241 fresh specimens from the upper respiratory tract.
- Data Source: Clinical sites.
- Annotation Protocol: Specimens were processed using bioMérieux NucliSENS® easyMag® for nucleic acid extraction. The ABI 7500 Fast Dx Instrument was used for amplification and detection with the Quidel Lyra™ Adenovirus Assay. Reference method: viral culture with an FDA-cleared composite method of direct fluorescent antibody staining (CDFA) followed by direct fluorescent antibody staining of the specimen (SDFA). A specimen was considered positive if either CDFA or SDFA was positive; negative if both were negative.
Retrospective Study:
- Sample Size: 105 frozen specimens from the upper respiratory tract.
- Data Source: Pediatric hospital in the Southwest United States.
- Annotation Protocol: Specimens tested concurrently with the Lyra™ Adenovirus Assay and an additional FDA-cleared molecular device.
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Precision/Reproducibility
- Study Type: Within Laboratory Repeatability and Reproducibility at three laboratory sites (two external, one in-house).
- Sample Size: Panels of four simulated samples (moderate positive, low positive, high negative, negative). For reproducibility, 90 results per level for each virus (triplicate testing x 2 operators x 3 replicates x 5 days x 3 sites).
- Key Results: The Lyra™ Adenovirus Assay produces highly reproducible results. Combined site data for reproducibility showed high rates of detection for positive panels (100% for Low and Moderate Positive, 86/89 for High Negative) and 0% for Negative panels, with low Coefficients of Variation (CVs) for Ct values.
Limit of Detection (LoD)
- Study Type: Analytical sensitivity study.
- Key Results: LoD ranged from $3.20 \times 10^{-2}$ TCID50/mL to $1.61 \times 10^{1}$ TCID50/mL depending on the HAdV species/serotype.
Analytical Reactivity (Inclusivity)
- Study Type: Inclusivity testing.
- Key Results: 49 HAdV serotypes were detected by the Lyra™ Adenovirus Assay at or near the species LoD. Three serotypes (38, 42, and 52) were evaluated using in silico analysis, showing high agreement of probes and primers with target regions.
Microbial Cross-Reactivity and Interference
- Study Type: Analytical specificity study.
- Key Results: No interference or cross-reactivity was observed with 30 viral, 26 bacterial, and 1 yeast strain tested.
Interfering Substances
- Study Type: Study to evaluate performance in the presence of potentially interfering/cross-reactive substances.
- Key Results: None of the eleven substances tested (e.g., mucin, blood, nasal sprays, lozenges, antiviral drugs) cross-reacted or interfered with the Lyra™ Adenovirus Assay.
Carry-Over and Cross-Contamination
- Study Type: Carry-over and cross-contamination study.
- Key Results: No cross-contamination or amplicon carry-over occurred.
Method Comparison (Clinical Performance)
- Type: Prospective Multi-Center Study
- Sample Size: 1239 fresh specimens.
- Key Results:
- Sensitivity: 100% (35/35; 95% CI: 90.1% to 100%)
- Specificity: 95.5% (1150/1204; 95% CI: 94.2% to 96.5%)
- 45 of 54 Lyra™ Adenovirus Assay positives that were negative by comparator were positive by an additional PCR assay.
- Type: Retrospective Study
- Sample Size: 105 frozen specimens.
- Key Results:
- Positive Percent Agreement: 100% (27/27; 95% CI: 87.5% to 100%)
- Negative Percent Agreement: 98.7% (77/78; 95% CI: 93.1% to 99.8%)
- The one positive by Lyra™ Adenovirus Assay that was negative by predicate was positive by a third FDA-cleared PCR assay.
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Prospective Study (vs. CDFA with SDFA):
- Sensitivity: 100% (35/35)
- Specificity: 95.5% (1150/1204)
Retrospective Study (vs. FDA-Cleared PCR Assay):
- Positive Percent Agreement: 100% (27/27)
- Negative Percent Agreement: 98.7% (77/78)
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.
Not Found
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
§ 866.3980 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
Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health and Human Services. The logo features a stylized caduceus symbol, which is often associated with medicine and healthcare. The words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" are arranged in a circular pattern around the caduceus symbol. The logo is simple, clean, and easily recognizable.
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
October 9, 2014
Quidel Corporation Ronald H. Lollar, Senior Director Clinical and Quality Affairs Diagnostic Hybrids, Inc. (Wholly Owned Subsidiary of Quidel) 2005 East State Street, Suite 100 Athens, OH 45701
Re: K141931
Trade/Device Name: Lyra™ Adenovirus Assay Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory viral panel multiplex nucleic acid assay Regulatory Class: II Product Code: OCC Dated: July 15, 2014 Received: July 16, 2014
Dear Mr. 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 (QS) 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,
Tamara V. Feldblyum -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) K141931
Device Name Lyra™ Adenovirus Assay
Indications for Use (Describe)
The Lyra™ Adenovirus Assay is a real-time polymerase chain reaction (PCR) in vitro diagnostic test for the qualitative detection of human adenovirus (HAdV) viral DNA isolated from nasal and nasopharyngeal swab specimens obtained from individuals exhibiting signs and symptoms of acute respiratory infections. The intended use of this test is to aid in the diagnosis of HAdV in conjunction with other clinical and laboratory findings.
The test detects, but does not differentiate. HAdV species (A, B, C, D, E, and F) or serotypes (HAdV 1-52).
Negative results do not preclude HAdV infection and should not be used as the sole basis for diagnosis, treatment or other patient management decisions.
Type of Use (Select one or both, as applicable)
| Prescription Use (Part 21 CFR 201 Subpart D) |
|---|
| Over-The-Counter Use (21 CFR 330 Subpart D) |
|X 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."
3
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 740-593-8437 – Fax lollar@dhiusa.com
Date of preparation of 510(k) summary:
July 15, 2014
Device Name:
Trade name - Lyra™ Adenovirus Assay Classification name - Respiratory viral panel multiplex nucleic acid assay Product Code - OCC Regulation Section - 21CFR 866.3980
Substantial Equivalency
The Lyra™ Adenovirus Assay used an FDA-cleared composite method of direct fluorescent antibody (DFA) staining of viral culture followed by DFA staining of the specimen as the reference comparator. The predicate device for the assay is the Adenovirus R-gene® US. The characteristics of the Lyra™ Adenovirus Assay ("Subject Device") and the legally marketed device are described in the Table below:
4
| Item | Subject Device
LyraTM Adenovirus
Assay | Predicate Device
(K121942)
Adenovirus R-gene
US Assay |
|------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | The LyraTM Adenovirus
Assay is a real-time
polymerase chain
reaction (PCR) in vitro
diagnostic test for the
qualitative detection of
human adenovirus
(HAdV) viral DNA
isolated from nasal and
nasopharyngeal swab
specimens obtained from
individuals exhibiting
signs and symptoms of
acute respiratory
infection. The intended
use of this test is to aid
in the diagnosis of
HAdV in conjunction
with other clinical and
laboratory findings.
The test detects, but does
not differentiate, HAdV
species (A, B, C, D, E,
and F) or serotypes
(HAdV 1-52).
Negative results do not
preclude HAdV
infection and should not
be used as the sole basis
for treatment or other
patient management
decisions. | Adenovirus R-gene US
is a Real Time PCR in
vitro diagnostic test for
the rapid and
qualitative detection of
Adenovirus viral DNA
isolated and purified
from nasopharyngeal
specimens (swab or
wash/aspirate) obtained
from individuals
exhibiting signs and
symptoms of acute
respiratory infection.
The intended use for
this test is to aid in the
diagnosis of
Adenovirus infections
in humans.
Negative results do not
preclude Adenovirus
infection and should
not be used as the sole
basis for treatment or
other management
decisions. |
| DNA
Amplification
Technology | Real-time polymerase
chain reaction (PCR) | Same |
5
| Item | Subject Device
Lyra™ Adenovirus
Assay | Predicate Device
(K121942)
Adenovirus R-gene
US Assay | | | |
|--------------------------------------|-------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------|--|--|--|
| Assay Results | Qualitative | Same | | | |
| Viral Target
Sequence | Penton base gene | Hexon gene | | | |
| Sample Types | Nasal and
Nasopharyngeal swabs | Nasopharyngeal swabs
and Nasopharyngeal
aspirates/washes | | | |
| Amplification | Self-contained and
automated | Same | | | |
| Detection
Techniques | Self-contained and
automated | Same | | | |
| Nucleic Acid
Extraction
Method | bioMérieux NucliSENS
easyMAG System | Same | | | |
| Collection and
Transport Media | Universal Transport
Medium (Copan/DHI),
MicroTest M4, M4-RT,
M5, or M6 Transport
Medium (Remel) | Universal Transport
Medium (DHI)
MicroTest M4RT
Transport (Remel) | | | |
| Instrument/Assay
Platform | Applied Biosystems
(ABI) 7500 Fast Dx
Real-Time PCR
Instrument | Cepheid SmartCycler
II | | | |
| Controls
Included with
Assav | Internal process control
(MS2 Bacteriophage;
Zeptometrix ) | Positive control
(Plasmid DNA)
Negative Control
(molecular grade
water)
Internal control (IC2)
phage particle | | | |
6
Intended Use
The Lyra™ Adenovirus Assay is a real-time polymerase chain reaction (PCR) in vitro diagnostic test for the qualitative detection of human adenovirus (HAdV) viral DNA isolated from nasal and nasopharyngeal swab specimens obtained from individuals exhibiting signs and symptoms of acute respiratory infections. The intended use of this test is to aid in the diagnosis of HAdV in conjunction with other clinical and laboratory findings.
The test detects, but does not differentiate, HAdV species (A, B, C, D, E, and F) or serotypes (HAdV 1-52).
Negative results do not preclude HAdV infection and should not be used as the sole basis for diagnosis, treatment or other patient management decisions
Methodology
The assay detects viral nucleic acids that have been extracted from a patient sample. A real-time PCR reaction is carried out using the ABI 7500 Fast Dx Real-Time PCR Instrument ("ABI Fast Dx Instrument") under optimized conditions in a single tube generating amplicons for adenovirus and the Process Control (PRC). Identification of human adenovirus (HAdV) and the PRC occurs by the use of target-specific primers and fluorescent-labeled probes that hybridize to conserved regions in the genomes of adenovirus and the PRC.
| Table 1. Lyra™ Probe Labels | |
|---|---|
| Target | Dye |
| HAdV | FAM |
| PRC | Cy5 |
Performance Data
Precision/Reproducibility
Precision
The Precision/Within Laboratory Repeatability study was assessed using a panel of four (4) simulated samples that include moderate positive and low positive, high negative and negative. The four (4) member panel consisting of HAdV and negative samples were tested by two (2) operators, in triplicate for twelve (12) days.
| Table 2. Results Summary – Cts and Positivity | |||||||
|---|---|---|---|---|---|---|---|
| Platform | Target | Pos. Control | Ct Values | ||||
| 5X LoD | 2X LoD | 0.5X LoD | Neg. Matrix | Neg. Control | |||
| ABI 7500 | |||||||
| Fast Dx | Operator 1 Avg Ct | 20.8 | 25.5 | 26.6 | 29.7 | NEG | NEG |
| Operator 2 Avg Ct | 20.4 | 25.4 | 26.7 | 29.3 | NEG | NEG |
7
Quidel Corporation
| Table 2. | Results Summary – Cts and Positivity | ||||||
|---|---|---|---|---|---|---|---|
| Platform | Target | Ct Values | |||||
| Pos. | |||||||
| Control | 5X LoD | 2X LoD | 0.5X | ||||
| LoD | Neg. | ||||||
| Matrix | Neg. | ||||||
| Control | |||||||
| Positivity | 100% | 100% | 100% | 100% | 0% | 0% |
Section 05, 510(k) Summary
The Lyra™ Adenovirus Assay produces results that are highly reproducible.
Reproducibility
The reproducibility of the Lyra™ Adenovirus Assay was evaluated at three (3) laboratory sites (two external, one in-house). Reproducibility was assessed using a panel of four (4) simulated samples that include moderate positive and low positive, high negative and negative adenovirus samples. The panels and controls were processed and tested on the ABI 7500 Fast Dx Instrument. Panels and controls were tested at each site by 2 operators for 5 non-consecutive days (triplicate testing x 2 operators x 3 replicates x 5 days x 3 sites = 90 results per level for each virus). The LoD values were based on the values obtained in the LoD study.
| Table 3. | Reproducibility Results | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Panel | Site 1 | Site 2 | Site 3 | Combined Site Data | ||||||||
| Member | ||||||||||||
| ID | Rate of | |||||||||||
| Detection | AVE | |||||||||||
| Ct | % | |||||||||||
| CV | Rate of | |||||||||||
| Detection | AVE | |||||||||||
| Ct | % | |||||||||||
| CV | Rate of | |||||||||||
| Detection | AVE | |||||||||||
| Ct | % | |||||||||||
| CV | Rate of | |||||||||||
| Detection | AVE | |||||||||||
| Ct | %CV | |||||||||||
| HAdV | ||||||||||||
| High | ||||||||||||
| Negative | ||||||||||||
| (0.5x LoD) | 30/30 | 29.0 | 4.9 | 30/30 | 28.9 | 4.2 | 26/29* | 27.8 | 8.8 | 86/89* | 28.6 | 6.2 |
| HAdV | ||||||||||||
| Low | ||||||||||||
| Positive | ||||||||||||
| (2x LoD) | 30/30 | 26.7 | 2.3 | 30/30 | 26.2 | 3.0 | 30/30 | 26.1 | 9.3 | 90/90 | 26.4 | 5.9 |
| HAdV | ||||||||||||
| Moderate | ||||||||||||
| Positive | ||||||||||||
| (5x LoD) | 30/30 | 25.2 | 1.7 | 30/30 | 24.8 | 3.0 | 30/30 | 24.1 | 6.8 | 90/90 | 24.8 | 4.8 |
| HAdV | ||||||||||||
| Negative | ||||||||||||
| Nasal | ||||||||||||
| Matrix | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/90 | N/A | N/A |
| HAdV | ||||||||||||
| Positive | ||||||||||||
| Control | 30/30 | 20.5 | 2.5 | 30/30 | 18.2 | 3.9 | 30/30 | 17.1 | 3.7 | 90/90 | 18.6 | 8.5 |
| HAdV | ||||||||||||
| Negative | ||||||||||||
| Control | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/30 | N/A | N/A | 0/90 | N/A | N/A |
8
- One aliquot was removed from the analysis because it had an invalid PRC result. Three aliquots (from the same replicate) tested negative.
The data from the combined sites indicates that the Lyra™ Adenovirus Assay, on the ABI 7500 Fast Dx Instrument, generates reproducible results for the detection of adenovirus and the internal control.
Limit of Detection (LoD)
The analytical sensitivity (limit of detection or LoD) of the Lyra™ Adenovirus Assay was determined using quantified (TCID50/mL) stocks of representatives of the six (6) species of adenovirus diluted in a negative matrix. Testing was performed with three manufactured device lots. Analytical sensitivity (LoD) is defined as the lowest concentration at which 95% of all replicates tested positive.
| Table 4. Summary of LoD Study | ||
|---|---|---|
| Species/ Serotype | LoD | |
| (TCID50/mL) | Avg. Ct Value | |
| A/31 | $8.00 x10^{-2}$ | 28.5 |
| B/3 | $8.00 x10^{-2}$ | 28.3 |
| C/1 | $8.00 x10^{-2}$ | 28.1 |
| D/19 | $1.61 x10^{1}$ | 28.6 |
| E/4 | $1.00 x10^{0}$ | 27.7 |
| F/41 | $3.20 x10^{-2}$ | 28.9 |
Analytical Reactivity (Inclusivity)
To verify the Lyra™ Adenovirus Assay detects the fifty-two (52) known serotypes of HAdV, 49 serotypes were initially tested at or near the species LoD (see above). Higher concentrations were tested if the organism was not detected at the LoD. Each of the 49 serotypes tested was detected by the Lyra Adenovirus Assay. Three (3) of the serotypes (38, 42, and 52) were unavailable for testing with the device. These serotypes were evaluated using in silico analysis.
| Species | Serotype | Concentration
Tested
(TCID50/mL) | Multiple of
LoD
Detected |
|--------------------------------------------|----------|----------------------------------------|--------------------------------|
| A | HAdV-12 | 1.60 x10-1 | 2x LoD |
| A | HAdV-18 | 1.60 x10-1 | 2x LoD |
| B | HAdV-7 | 1.60 x10-1 | 2x LoD |
| Species | Serotype | Concentration
Tested
(TCID50/mL) | Multiple
LoD
Detected |
| | HAdV-11 | 1.60 x10-1 | 2x LoD |
| | HAdV-14 | 1.60 x10-1 | 2x LoD |
| | HAdV-16 | 1.60 x10-1 | 2x LoD |
| | HAdV-21 | 2.00 x101 | 250x LoD |
| | HAdV-34 | 1.60 x10-1 | 2x LoD |
| | HAdV-35 | 1.60 x10-1 | 2x LoD |
| | HAdV-50 | 1.60 x10-1 | 2x LoD |
| | HAdV-2 | 1.60 x10-1 | 2x LoD |
| C | HAdV-5 | 1.60 x10-1 | 2x LoD |
| | HAdV-6 | 1.60 x10-1 | 2x LoD |
| F | HAdV-40 | 6.40 x10-2 | 2x LoD |
| | HAdV-8 | 3.22 x10-1 | 2x LoD |
| | HAdV-9 | 3.22 x10-1 | 2x LoD |
| | HAdV-10 | 3.22 x10-1 | 2x LoD |
| | HAdV-13 | 3.22 x10-1 | 2x LoD |
| | HAdV-15 | 9.66 x101 | 6x LoDb |
| | HAdV-17 | 9.66 x101 | 6x LoDc |
| | HAdV-20 | 3.22 x10-1 | 2x LoD |
| | HAdV-22 | 3.22 x10-1 | 2x LoD |
| | HAdV-23 | 3.22 x10-1 | 2x LoD |
| | HAdV-24 | 3.22 x10-1 | 2x LoD |
| D | HAdV-25 | 3.22 x10-1 | 2x LoD |
| | HAdV-26 | 3.22 x10-1 | 2x LoD |
| | HAdV-27 | 3.22 x10-1 | 2x LoD |
| | HAdV-28 | 3.22 x10-1 | 2x LoD |
| | HAdV-29 | 3.22 x10-1 | 2x LoD |
| | HAdV-30 | 3.22 x10-1 | 2x LoD |
| | HAdV-32 | 3.22 x10-1 | 2x LoD |
| | HAdV-33 | 3.22 x10-1 | 2x LoD |
| | HAdV-36 | 3.22 x10-1 | 2x LoD |
| | HAdV-37 | 3.22 x10-1 | 2x LoD |
| Table 5. HAdV Serotypes Reactivity Summary | | | |
| Species | Serotype | Concentration
(TCID50/mL) | Multiple of
LoD
Detected |
| | HAdV-43 | 3.22 x10-1 | 2x LoD |
| | HAdV-44 | 3.22 x10-1 | 2x LoD |
| | HAdV-45 | 3.22 x10-1 | 2x LoD |
| | HAdV-46 | 3.22 x10-1 | 2x LoD |
| | HAdV-47 | 3.22 x10-1 | 2x LoD |
| | HAdV-48 | 3.22 x10-1 | 2x LoD |
| | HAdV-49 | 3.22 x10-1 | 2x LoD |
| | HAdV-51 | 3.22 x10-1 | 2x LoD |
9
10
4 HAdV-21 required repeat testing and was ultimately found to be detected at 250X LoD.
b HAdV-15 required repeat testing and was ultimately found to be detected at 6X LoD.
C HAdV-17 required repeat testing and was ultimately found to be detected at 6X LoD.
HAdV Species D serotypes 38 and 42 and Species G serotype 52 (often associated with conjunctivitis and gastroenteritis) were not evaluated for inclusivity as the sponsor could not obtain these strains. Instead, the manufacturer conducted an in silico analysis where they aligned their probe and primers with the target regions from a single sequence (obtained from GenBank) for each of these adenovirus types. The alignments showed a 100% agreement of the forward primer with types 38, 42, and 52. For the reverse primer, there were 3 mismatches detected on the sequences for types 38 and 42 (84% homology) and 2 mismatches with the sequence for type 52 (89.5% homology). For the probe, there was 100% agreement with types 38 and 42, while three mismatches were observed with the sequence for type 52 (85% homology).
Microbial Cross-Reactivity and Interference
The analytical specificity of the Lyra Adenovirus Assay was evaluated by testing a panel consisting of 30 viral, 26 bacterial, and 1 yeast strain representing respiratory pathogens or flora commonly present in the nasopharynx. The organisms were tested in the presence and absence of 2X LoD HAdV to determine if there was interference or cross-reactivity, respectively. Samples were extracted using the NucliSENS easyMAG instrument and tested in triplicate. All HAdV positive samples remained positive and all HAdV negative samples remained negative. indicating that there was no interference or cross-reactivity in the presence of the organisms tested. Each microorganism was tested three times, once in the presence of HAdV type 4, once in the presence of HAdV type 31, and once in the presence of negative nasal matrix.
11
| Table 6. Organisms used for the study were the following: | Concentration tested (TCID50/mL or CFU/mL) | Lyra Adenovirus Assay Result | |
|---|---|---|---|
| Organism | Interference (+ HAdV) | Cross-reactivity (- HAdV) | |
| Bordetella pertussis | 9.08E+08 | Positive | Negative |
| Bordetella bronchiseptica | 5.40E+08 | Positive | Negative |
| Chlamydophila pneumonia | 0.11 ng/ µL | Positive | Negative |
| Chlamydia trachomatis | 2.10E+06 | Positive | Negative |
| Legionella pneumophila | 1.42E+09 | Positive | Negative |
| Mycobacterium intracellualre | 1.53E+09 | Positive | Negative |
| Mycobacterium tuberculosis | 9.30E+06 | Positive | Negative |
| Mycobacterium avium | 3.18E+09 | Positive | Negative |
| Mycoplasma pneumoniae | 3.16E+06 | Positive | Negative |
| Haemophilus influenzae | 4.00E+08 | Positive | Negative |
| Pseudomonas aeruginosa | 1.32E+09 | Positive | Negative |
| Proteus vulgaris | 6.53E+08 | Positive | Negative |
| Proteus mirabilis | 1.19E+09 | Positive | Negative |
| Neisseria gonorrhoeae | 1.40E+09 | Positive | Negative |
| Neisseria meningitidis | 1.29E+08 | Positive | Negative |
| Neisseria mucosa | 1.61E+08 | Positive | Negative |
| Klebsiella pneumoniae | 9.75E+08 | Positive | Negative |
| Escherichia coli | 1.13E+09 | Positive | Negative |
| Moraxella catarrhalis | 1.26E+09 | Positive | Negative |
| Corynebacterium diptheriae | 3.44E+08 | Positive | Negative |
| Lactobacillus plantarum | 3.18E+08 | Positive | Negative |
| Streptococcus pneumoniae | 1.43E+08 | Positive | Negative |
| Streptococcus pyogenes | 6.38E+08 | Positive | Negative |
| Streptococcus salivarius | 5.40E+08 | Positive | Negative |
| Staphylococcus epidermidis | 9.23E+08 | Positive | Negative |
| Staphylococcus aureus | 6.08E+08 | Positive | Negative |
| Candida albican | 9.70E+07 | Positive | Negative |
| Coronavirus 229E | 2.46E+07 | Positive | Negative |
| Coronavirus NL63 | 1.41E+04 | Positive | Negative |
| Coronavirus OC43 | 2.42E+07 | Positive | Negative |
| Coxsackievirus B4 | 2.00E+07 | Positive | Negative |
| Coxsackievirus B5/10/2006 | 3.62E+05 | Positive | Negative |
| Cytomegalovirus | 2.14E+06 | Positive | Negative |
| Table 6. Organisms used for the study were the following: | |||
| Lyra Adenovirus Assay Result | |||
| Organism | Concentration tested | ||
| (TCID50/mL or | |||
| CFU/mL) | Interference | ||
| (+ HAdV) | Cross- | ||
| reactivity | |||
| (- HAdV) | |||
| Echovirus 6 | 7.60E+08 | Positive | Negative |
| Echovirus 7 | 4.45E+06 | Positive | Negative |
| Echovirus 9 | 2.17E+07 | Positive | Negative |
| Echovirus 11 | 2.17E+05 | Positive | Negative |
| Enterovirus 70 | 2.41E+06 | Positive | Negative |
| Enterovirus 71 | 2.03E+05 | Positive | Negative |
| Epstein Barr Virus | 9.27E+07 | Positive | Negative |
| HSV Type 1 MacIntyre Strain | 5.89E+06 | Positive | Negative |
| HSV Type 2 Strain G | 1.96E+06 | Positive | Negative |
| Human Metapneumovirus (A1) | 3.66E+05 | Positive | Negative |
| Human Rhinovirus 45 | 2.94E+04 | Positive | Negative |
| Human Rhinovirus 52 | 2.63E+04 | Positive | Negative |
| Influenza A/Mexico/4108/2009 | 4.08E+05 | Positive | Negative |
| Influenza A/Port Chalmers | 3.55E+08 | Positive | Negative |
| Influenza B/Florida/04/2006 | 1.54E+06 | Positive | Negative |
| Measles | 1.95E+06 | Positive | Negative |
| Mumps Virus | 2.75E+08 | Positive | Negative |
| Parainfluenza Type 1 | 3.97E+06 | Positive | Negative |
| Parainfluenza Type 2 | 3.15E+08 | Positive | Negative |
| Parainfluenza Type 3 | 2.36E+07 | Positive | Negative |
| Parainfluenza Type 4A | 1.04E+05 | Positive | Negative |
| RSV A (Long) | 4.36E+04 | Positive | Negative |
| RSV B (Wash/18537/62) | 3.43E+05 | Positive | Negative |
| Varicella Zoster Virus | 1.11E+04 | Positive | Negative |
12
Interfering Substances
A study was performed on the ABI 7500 Fast Dx Instrument to evaluate the performance of the Lyra™ Adenovirus Assay in the presence of eleven (11) of potentially interfering/cross-reactive substances, at clinically relevant levels, that might be present in specimens. Each substance was tested in the presence of 2X LoD HAdV and with negative matrix.
13
| Table 7.
| Interfering/Cross-reactive Substances Summary | |||||||
|---|---|---|---|---|---|---|---|
| HAdV | |||||||
| Substance Name | Species E | ||||||
| Serotype 4 | Species A | ||||||
| Serotype 31 | No Analyte | Inhibition | |||||
| (yes/no) | |||||||
| Ct Avg. | SD | Ct Avg. | SD | Ct Avg. | SD | ||
| Controls | 26.0 | 0.6 | 27.6 | 0.9 | NEG | N/A | N/A |
| Mucin (Bovine Submaxillary | |||||||
| Gland, type I-S) | 26.7 | 0.8 | 27.1 | 0.2 | NEG | N/A | No |
| Blood (human), EDTA | |||||||
| anticoagulated | 28.0 | 1.2 | 27.6 | 0.6 | NEG | N/A | No |
| Neo-Synephrine | 27.2 | 1.0 | 26.9 | 0.2 | NEG | N/A | No |
| Afrin Nasal Spray | 26.2 | 0.8 | 26.7 | 0.1 | NEG | N/A | No |
| Zicam Homeopathic Non- | |||||||
| Drowsy Allergy Relief No | |||||||
| Drip Liquid Nasal Gel | 25.6 | 0.3 | 26.9 | 0.3 | NEG | N/A | No |
| Saline Nasal Spray | 25.8 | 0.5 | 28.2 | 1.3 | NEG | N/A | No |
| OTC Throat Lozenges: | |||||||
| Ricola Action Cherry | 26.0 | 0.3 | 26.3 | 1.3 | NEG | N/A | No |
| Zanamivir | 25.8 | 0.8 | 26.5 | 0.3 | NEG | N/A | No |
| Tobramycin | 26.6 | 0.2 | 26.8 | 0.1 | NEG | N/A | No |
| Mupirocin | 26.4 | 0.4 | 27.7 | 0.8 | NEG | N/A | No |
| Oseltamivir phosphate | 26.0 | 0.0 | 27.2 | 0.2 | NEG | N/A | No |
None of the eleven (11) of substances tested cross-reacts with the Lyra™ Adenovirus Assay.
Carry-Over and Cross-Contamination
Studies were performed on the ABI 7500 Fast Dx Instrument using a 96-sample panel consisting of 48 high positive (1.0 x 10 TCID50 mL HAdV type 4) and 48 negative specimens (negative nasal matrix). The high positive samples were extracted and added to the plate in a checkerboard pattern that alternated with the negative samples. This testing was repeated over a 5-day period.
Over the course of 5 days, cross-contamination and amplicon carry-over did not occur with the Lyra™ Adenovirus Assay when extracted using the NucliSens easyMAG automated nucleic acid extraction instrument and analyzed on the ABI 7500 Fast Dx Instrument.
14
Method Comparison
Prospective Study
The evaluation of the Lyra™ Adenovirus Assay occurred in two separate studies: a prospective multi-center study using one thousand two hundred and forty-one (1241) fresh specimens from the upper respiratory tract; and a retrospective study using one hundred five (105) frozen specimens from the upper respiratory tract. In both studies the specimens were processed the bioMérieux NucliSENS® easyMag® at all sites for the extraction of nucleic acids from the clinical specimens. The ABI 7500 Fast Dx Instrument was used with the Quidel Lyra™ Adenovirus Assay for the amplification and detection of the target nucleic acids. The prospective specimens were also processed and tested with viral culture with an FDA-cleared composite method of direct fluorescent antibody staining (CDFA) followed by direct fluorescent antibody staining of the specimen (SDFA). A specimen was called positive if either the CDFA or the SDFA were positive. For a specimen to be called negative, it must be negative for both CDFA and SDFA. The retrospective specimens were extracted and tested with an additional FDAcleared molecular assay.
| Age | # Tested | Lyra
Adenovirus
Assay Positive | Expected
Value | Total Positive
by Reference
Method | Observed
Prevalence |
|----------|----------|--------------------------------------|-------------------|------------------------------------------|------------------------|
| 21 | 437 | 4 | 0.9% | 3 | 0.7% |
| Total | 1241 | 89 | 7.2% | 35 | 2.8% |
Expected Values for the Winter of 2013 and 2014 (Combined)
One thousand two hundred forty-one (1241) fresh specimens were collected and transported to each laboratory for testing with the Lyra™ Adenovirus Assay. Two (2) specimens were invalid when initially tested with the Lyra™ Adenovirus Assay. The specimens were re-tested according to the instructions for use and were invalid upon repeat testing. The specimens have been removed from the data presented below. The specimens shipped daily with cold packs for CDFA and SDFA to the central location and were tested within 72-hours of collection. The table below details the HAdV results for the remaining one thousand two hundred thirty-nine (1239) specimens.
15
| Combined Site Data | |||
|---|---|---|---|
| Comparator: CDFA with SDFA | |||
| LyraTM Adenovirus | |||
| Assay | Positive | Negative | Total |
| Positive | 35 | 54* | 89 |
| Negative | 0 | 1150 | 1150 |
| Total | 35 | 1204 | 1239 |
| 95% CI | |||
| Sensitivity | 35/35 | 100% | 90.1% to 100% |
| Specificity | 1150/1204 | 95.5% | 94.2 % to 96.5% |
- Forty-five (45) of the fifty-four (54) positives were positive by an additional PCR assay. Four (4) of the fifty-four (54) positives were negative by an additional PCR assay. Two (2) of the fifty-four (54) positives were invalid by an additional PCR assay. Three (3) of the fifty-four (54) positives had insufficient volume for testing by an additional PCR assay.
Retrospective Study
Due to the low prevalence of HAdV at the clinical sites during the study period, a retrospective study was conducted at Site 1 with specimens obtained from a pediatric hospital in the Southwest United States. One hundred five (105) frozen specimens from the upper respiratory tract were tested concurrently with the Lyra™ Adenovirus Assay and an additional FDA-cleared molecular device.
| Adenovirus | |||
|---|---|---|---|
| Comparator: FDA-Cleared PCR Assay | |||
| Lyra™ Adenovirus Assay | Positive | Negative | Total |
| Positive | 27 | 1* | 28 |
| Negative | 0 | 77 | 77 |
| Total | 27 | 78 | 105 |
| 95% CI | |||
| Positive Percent Agreement | 27/27 | 100% | 87.5% to 100% |
| Negative Percent Agreement | 77/78 | 98.7% | 93.1% to 99.8% |
- One (1) of one (1) positive was positive by a third FDA-cleared PCR assay.
Statement of Safety and Effectiveness
In prospectively collected clinical specimens, when performed on the ABI 7500 Fast Dx Instrument, the Lyra™ Adenovirus Assay vielded good sensitivity and specificity when compared to the composite reference method of direct specimen fluorescent antibody (SDFA) and viral culture with DFA (CDFA).
16
In retrospectively collected clinical specimens, when performed on the ABI 7500 Fast Dx Instrument, the Lyra™ Adenovirus Assay yielded good positive and negative percent agreement when compared to a FDA-cleared molecular device.