(96 days)
No
The device description details a standard PCR-based assay with analysis software that interprets fluorescence data based on melting temperatures. There is no mention of AI or ML algorithms being used for data analysis or interpretation.
No
The device is described as an "in vitro diagnostic test" for the detection and typing of HSV DNA, which is used as an aid in diagnosis, not for treatment.
Yes
Explanation: The "Intended Use / Indications for Use" section explicitly states that the device is "a polymerase chain reaction (PCR)-based qualitative in vitro diagnostic test for the detection and typing of herpes simplex virus (HSV1&2) DNA... as an aid in the diagnosis of genital herpes infection."
No
The device is a kit that includes reagents and relies on specific hardware (Roche MagNA Pure LC instrument, Roche LightCycler 1,2 real-time PCR instrument) for sample processing and analysis, in addition to the analysis software.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use/Indications for Use: The very first sentence explicitly states: "The MultiCode®-RTx HSV 1&2 Kit is a polymerase chain reaction (PCR)-based qualitative in vitro diagnostic test for the detection and typing of herpes simplex virus (HSV1&2) DNA in vaginal lesions." This directly identifies it as an IVD.
- Device Description: The description details how the test is performed on patient specimens (vaginal swab specimens) in a laboratory setting, using reagents and instruments to analyze biological samples outside of the body. This is the core characteristic of an in vitro diagnostic device.
- Performance Studies: The document describes clinical performance studies conducted to evaluate the device's accuracy in detecting and typing HSV in patient samples, which is a standard requirement for IVD devices.
- Predicate Device: The mention of a predicate device (K971662; Diagnostic Hybrids, Inc. ELVIS® HSV ID/Typing Test System) further confirms its classification as an IVD, as predicate devices are used for comparison in the regulatory clearance process for new IVDs.
N/A
Intended Use / Indications for Use
The MultiCode®-RTx HSV 1&2 Kit is a polymerase chain reaction (PCR)-based qualitative in vitro diagnostic test for the detection and typing of herpes simplex virus (HSV1&2) DNA in vaginal lesions. It is indicated for use in the detection and typing of HSV-1 or HSV-2 in vaginal lesion swab specimens from symptomatic female patients as an aid in the diagnosis of genital herpes infection.
Warning: The device is not FDA cleared for the use with cerebral spinal fluid (CSF) or any lesions other than vaginal. This assay is not intended to be used for male penile specimens, for prenatal screening, or for females under the age of 18 years.
Product codes
OQO
Device Description
The EraGen MultiCode®-RTx Herpes Simplex Virus 1 & 2 kit is a polymerase chain reaction (PCR)-based qualitative in vitro diagnostic test for the detection and typing of herpes simplex virus (HSV) DNA using vaginal swab specimens.
Patient vaginal swab specimens are collected in Copan Universal Transport Medium, or identical Copan manufactured media formulations (Becton Dickinson Universal Viral Transport Media, Copan branded Universal Transport Medium for LabCorp, and the Quest Viral Culture Media) and transported to the laboratory. An extractable sample processing control (SPC) target is added to the specimen prior to lysis. The SPC controls for specimen lysis, for recovery of extracted nucleic acid, for inhibitory substances and for PCR reagent and instrument integrity. The specimen is lysed and nucleic acid is extracted using the Roche MagNA Pure LC instrument using the Roche MagNA Pure LC Total Nucleic Acid Isolation Kit.
A sample of the extracted nucleic acid is added to the MultiCode® RTx Herpes Simplex Virus 1 & 2 Kit reagents that contain a primer pair specific to HSV-1 and HSV-2 and a second primer pair specific to the SPC sequence. The two specific primer pairs are labeled with distinct fluorophore labels. PCR amplification is performed and assay fluorescence is monitored using the Roche LightCycler 1,2 real-time PCR instrument. Incorporation of the quencher-labeled nucleotide causes a decrease in assay fluorescence. Following amplification, the reaction is slowly heated and fluorescence is monitored. The strands of the amplification products will separate at a specific melting temperature (Tm) that is determined by an increase in fluorescence as the strands are separated. The instrument fluorescence output is analyzed and test results are determined using the MultiCode -RTx Herpes Simplex Virus 1 & 2 Kit Analysis Software. A printed results report is generated.
The following reagents, software and instructions for use are supplied with the MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit:
- HSV-1&2 Reaction Buffer.
- HSV-1&2 Primer Mix.
- HSV-1 Positive Control.
- HSV-2 Positive Control
- Sample Processing Control.
- Nuclease Free Water.
- Analysis Software.
- Certificate of Analysis
- Package Insert.
The MultiCode -RTx Herpes Simplex Virus 1 & 2 Kit reagents are intended to be stored at -15℃ to -30℃ and are stable until the expiration date indicated on the box.
The following ancillary reagents are required to perform the test but are not supplied:
- Specific lots of the ancillary reagent Clontech TITANIUM® Tag DNA Polymerase have been qualified for use with the MultiCode®-RTx HSV 1&2 Kit by EraGen Bioscience.
- The MultiCode®-RTx HSV 1&2 Kit product performance NOTE: requires that only qualified manufacturer lots of Clontech TITANIUM® Taq DNA Polymerase be used with the device. Any lots not specifically qualified by EraGen Bioscience for use with the MultiCode®-RTx HSV 1&2 Kit are not validated for use with this assay, and may cause erroneous results.
To find an up to date list of Qualified Clontech TITANIUM® Taq DNA Polymerase go to the EraGen Bioscience website Support page support(@eragen.com or call Customer Support at (866) 327-3290.
Use ancillary reagents only with the instructions for use contained here in this package insert. Discard any instructions for use that may be packaged with these ancillary reagents.
Any assay problems or failures that are suspected to involve the Clontech TITANIUM® Tag DNA Polymerase should be reported to EraGen Biosciences, Inc.
- The MultiCode®-RTx HSV 1&2 Kit product performance NOTE: requires that only qualified manufacturer lots of Clontech TITANIUM® Taq DNA Polymerase be used with the device. Any lots not specifically qualified by EraGen Bioscience for use with the MultiCode®-RTx HSV 1&2 Kit are not validated for use with this assay, and may cause erroneous results.
- Copan Universal Transport Medium, or identical Copan manufactured media formulations (Becton Dickinson Universal Viral Transport Media, Copan branded Universal Transport Medium for LabCorp, and the Ouest Viral Culture Media) - for use as the Kit Negative Control
- Roche MagNA Pure LC Total Nucleic Acid Isolation Kit
Ancillary reagents are only to be used in accordance with the MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit package insert.
The following equipment and supplies are required to perform the test, but are not supplied:
- Roche MagNA Pure LC Instrument
- Roche LightCycler version 1.2 instrument with LightCycler software version 3.5
- LightCycler Carousel Centrifuge
- LightCycler Capillaries
- LightCycler Centrifuge Adapters in Cooling Block
- Vortex
- Minicentrifuge
- Appropriate sized pipetters and associated aerosol-barrier tips
- Appropriate sized screw cap tubes and caps
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Vaginal lesions
Indicated Patient Age Range
Not intended for females under the age of 18 years.
Intended User / Care Setting
Clinical laboratories
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
A prospective study was performed at three U.S. clinical laboratories during 2008-2009. Vaginal lesion swab specimens were sent to the clinical laboratory for routine Herpes Simplex Virus testing. The reference method was a cell culture based ELVIS® Herpes Simplex Virus identification and typing test. A total of 1041 vaginal swab specimens were tested by the MultiCode -RTx HSV 1&2 Kit and by the reference method.
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Analytical Performance
- Precision/Reproducibility: Evaluated at three U.S. clinical laboratories. A panel was prepared containing six simulated HSV-1 and HSV-2 samples (High Negative, Low Positive, and High Positive samples). The panel, along with external HSV-1 and HSV-2 positive and negative controls, was assayed in triplicate. Kit positive and negative controls were included with each assay run. Panels and controls were tested at each site by 2 operators, 1 time each per day for 5 days (N = 900).
- Total agreement with expected results for HSV-1 Positive Control, HSV-2 Positive Control, HSV-1/HSV-2 Negative Control, HSV-1 High Negative, HSV-2 High Negative, HSV-2 Low Positive, HSV-2 High Positive was 100%.
- Total agreement with expected results for PN 1750 HSV-1 Positive External Control and PN 1751 HSV-2 Positive External Control was 98.9%.
- Total agreement with expected results for PN 1754 HSV-1/HSV-2 Negative External Control and HSV-1 High Positive was 100%.
- Total agreement with expected results for HSV-1 Low Positive was 98.9%.
- Limit of Detection (LoD) / Limit of Blank (LoB): Study performed at EraGen Biosciences using quantified (TCID50/mL) cultures of HSV-1 and HSV-2 serially diluted in Copan Universal Transport Media. Each viral strain was extracted and tested in replicates of 60 per concentration.
- LoD for HSV-1 was 2.0 x 10^3 TCID50/mL (100% detection, 95% CI: 93.94 - 100%).
- LoD for HSV-2 was 6.4 x 10^1 TCID50/mL (98.3% detection, 95% CI: 91.06 – 99.96%).
- LoB for HSV-1 was 2.5 x 10^2 TCID50/mL (3.33% detected, 95% CI: 0.41 – 11.53%).
- LoB for HSV-2 was 4.0 TCID50/mL (0.00% detected, 95% CI: 0.00 - 5.96%).
- Analytical Specificity and Cross Reactivity: Evaluated at EraGen Biosciences using a panel of 22 different organisms. No HSV positive results were observed for any of the organisms tested. No interference was observed when the panel was spiked into HSV-1 and HSV-2 near the LoD.
- Interfering Substances: Evaluated at EraGen Biosciences using a panel of 6 substances. No interference was observed in the presence of exogenous and endogenous substances.
- Competitive Inhibition: Evaluated internally using simulated samples with varying concentrations of HSV-1 and HSV-2.
- Highest concentration of co-infecting target (HSV-2) that can be present while maintaining 95% detection of 3X LoD HSV-1 target was 1X LoD.
- Highest concentration of co-infecting target (HSV-1) that can be present while maintaining 95% detection of 3X LoD HSV-2 target was 1X LoD.
- No competitive inhibition observed when HSV-1 and HSV-2 were at equal concentrations (5X LoD to 500X LoD).
- Carry-over/Contamination: Evaluated internally with HSV-1 target. No carry-over/cross-contamination observed.
Clinical Performance:
- Prospective Study: Performed at three U.S. clinical laboratories (2008-2009). 1041 vaginal swab specimens were tested.
- HSV-1 Comparison Results:
- N=1041 total samples.
- Sequence analysis detected HSV-1 in 12 of 16 MultiCode-RTx positive/Reference Method negative discordant samples.
- Sequence analysis detected HSV-1 in 1 of 8 MultiCode-RTx negative/Reference Method positive discordant samples.
- HSV-2 Comparison Results:
- N=1041 total samples.
- Sequence analysis detected HSV-2 in 43 of 53 MultiCode-RTx positive/Reference Method negative discordant samples.
- Sequence analysis detected HSV-2 in 2 of 10 MultiCode-RTx negative/Reference Method positive discordant samples.
- HSV-1 Comparison Results:
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Herpes Simplex Virus Type 1 Comparison Results
- Sensitivity: 92.4% (95% Confidence Interval: 85.7 - 96.1%)
- Specificity: 98.3% (95% Confidence Interval: 97.2 - 98.9%)
Herpes Simplex Virus Type 2 Comparison Results
- Sensitivity: 95.2% (95% Confidence Interval: 91.4 – 97.4%)
- Specificity: 93.6% (95% Confidence Interval: 91.8 – 95.1%)
Predicate Device(s)
Reference Device(s)
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information
Not Found
§ 866.3305 Herpes simplex virus serological assays.
(a)
Identification. Herpes simplex virus serological assays are devices that consist of antigens and antisera used in various serological tests to identify antibodies to herpes simplex virus in serum. Additionally, some of the assays consist of herpes simplex virus antisera conjugated with a fluorescent dye (immunofluorescent assays) used to identify herpes simplex virus directly from clinical specimens or tissue culture isolates derived from clinical specimens. The identification aids in the diagnosis of diseases caused by herpes simplex viruses and provides epidemiological information on these diseases. Herpes simplex viral infections range from common and mild lesions of the skin and mucous membranes to a severe form of encephalitis (inflammation of the brain). Neonatal herpes virus infections range from a mild infection to a severe generalized disease with a fatal outcome.(b)
Classification. Class II (special controls). The device is classified as class II (special controls). The special control for the device is FDA's revised guidance document entitled “Class II Special Controls Guidance Document: Herpes Simplex Virus Types 1 and 2 Serological Assays.” For availability of the guidance revised document, see § 866.1(e).
0
MAY 1 2 2010
510(k) Summary MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit
1.0 Submitter
EraGen Biosciences, Inc. 918 Deming Way Suite 201 Madison, Wisconsin 53717 Telephone: (608) 662-9000
Establishment Registration Number: 3003232610
Contact Person
Randal Vader Vice President, Quality, Regulatory and Clinical Affairs Telephone: (608) 692-7839
Date of Summary Preparation
February 1, 2010
2.0 Device Identification
| Product Name: | MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit |
|---|---|
| Common Name: | Herpes Simplex Virus 1 and 2 Real-Time PCR Assay |
| Classification: | Class II |
| Product Code: | OQO |
| Regulation Number: | 21 CFR 866.3305 |
3.0 Device to Which Substantial Equivalence is Claimed
Diagnostic Hybrids, Inc. ELVIS® HSV ID/Typing Test System 510(k) Number: K971662
1
4.0 Description of Device
The EraGen MultiCode® -RTx Herpes Simplex Virus 1 & 2 kit is a polymerase chain reaction (PCR)-based qualitative in vitro diagnostic test for the detection and typing of herpes simplex virus (HSV) DNA using vaginal swab specimens.
Patient vaginal swab specimens are collected in Copan Universal Transport Medium, or identical Copan manufactured media formulations (Becton Dickinson Universal Viral Transport Media, Copan branded Universal Transport Medium for LabCorp, and the Quest Viral Culture Media) and transported to the laboratory. An extractable sample processing control (SPC) target is added to the specimen prior to lysis. The SPC controls for specimen lysis, for recovery of extracted nucleic acid, for inhibitory substances and for PCR reagent and instrument integrity. The specimen is lysed and nucleic acid is extracted using the Roche MagNA Pure LC instrument using the Roche MagNA Pure LC Total Nucleic Acid Isolation Kit.
A sample of the extracted nucleic acid is added to the MultiCode® RTx Herpes Simplex Virus 1 & 2 Kit reagents that contain a primer pair specific to HSV-1 and HSV-2 and a second primer pair specific to the SPC sequence. The two specific primer pairs are labeled with distinct fluorophore labels. PCR amplification is performed and assay fluorescence is monitored using the Roche LightCycler 1,2 real-time PCR instrument. Incorporation of the quencher-labeled nucleotide causes a decrease in assay fluorescence. Following amplification, the reaction is slowly heated and fluorescence is monitored. The strands of the amplification products will separate at a specific melting temperature (Tm) that is determined by an increase in fluorescence as the strands are separated. The instrument fluorescence output is analyzed and test results are determined using the MultiCode -RTx Herpes Simplex Virus 1 & 2 Kit Analysis Software. A printed results report is generated.
The following reagents, software and instructions for use are supplied with the MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit:
- HSV-1&2 Reaction Buffer .
- HSV-1&2 Primer Mix .
- HSV-1 Positive Control .
- HSV-2 Positive Control �
- Sample Processing Control .
- Nuclease Free Water . .
- Analysis Software .
2
- . Certificate of Analysis
- Package Insert .
The MultiCode -RTx Herpes Simplex Virus 1 & 2 Kit reagents are intended to be stored at -15℃ to -30℃ and are stable until the expiration date indicated on the box.
The following ancillary reagents are required to perform the test but are not supplied:
- . Specific lots of the ancillary reagent Clontech TITANIUM® Tag DNA Polymerase have been qualified for use with the MultiCode®-RTx HSV 1&2 Kit by EraGen Bioscience.
- The MultiCode®-RTx HSV 1&2 Kit product performance NOTE: requires that only qualified manufacturer lots of Clontech TITANIUM® Taq DNA Polymerase be used with the device. Any lots not specifically qualified by EraGen Bioscience for use with the MultiCode®-RTx HSV 1&2 Kit are not validated for use with this assay, and may cause erroneous results.
To find an up to date list of Qualified Clontech TITANIUM® Taq DNA Polymerase go to the EraGen Bioscience website Support page support(@eragen.com or call Customer Support at (866) 327-3290.
Use ancillary reagents only with the instructions for use contained here in this package insert. Discard any instructions for use that may be packaged with these ancillary reagents.
Any assay problems or failures that are suspected to involve the Clontech TITANIUM® Tag DNA Polymerase should be reported to EraGen Biosciences, Inc.
- Copan Universal Transport Medium, or identical Copan manufactured . media formulations (Becton Dickinson Universal Viral Transport Media, Copan branded Universal Transport Medium for LabCorp, and the Ouest Viral Culture Media) - for use as the Kit Negative Control
- . Roche MagNA Pure LC Total Nucleic Acid Isolation Kit
Ancillary reagents are only to be used in accordance with the MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit package insert.
3
The following equipment and supplies are required to perform the test, but are not supplied:
- . Roche MagNA Pure LC Instrument
- Roche LightCycler version 1.2 instrument with LightCycler software . version 3.5
- LightCycler Carousel Centrifuge .
- LightCycler Capillaries .
- . LightCycler Centrifuge Adapters in Cooling Block
- Vortex .
- . Minicentrifuge
- Appropriate sized pipetters and associated aerosol-barrier tips .
- Appropriate sized screw cap tubes and caps .
5.0 Indications for Use
The MultiCode® -RTx HSV 1&2 Kit is a polymerase chain reaction (PCR)-based qualitative in vitro diagnostic test for the detection and tvping of herpes simplex virus (HSV1&2) DNA in vaginal lesions. It is indicated for use in the detection and typing of HSV-1 or HSV-2 in vaginal lesion swab specimens from symptomatic female patients as an aid in the diagnosis of genital herpes infection.
Warning: The device is not FDA cleared for the use with cerebral spinal fluid (CSF) or any lesions other than vaginal. This assay is not intended to be used for male penile specimens, for prenatal screening, or for females under the age of 18 years.
6.0 Intended Use
The MultiCode® -RTx HSV 1&2 Kit is a polymerase chain reaction (PCR)-based qualitative in vitro diagnostic test for the detection and typing of herpes simplex virus (HSV1&2) DNA in vaginal lesions. It is indicated for use in the detection and typing of HSV-1 or HSV-2 in vaginal lesion swab specimens from symptomatic female patients as an aid in the diagnosis of genital herpes infection.
Warning: The device is not FDA cleared for the use with cerebral spinal fluid (CSF) or any lesions other than vaginal. This assay is not intended to be used for male penile specimens, for prenatal screening, or for females under the age of 18 years.
4
7.0 Comparison of the New Device with the Predicate Device
The EraGen Bioscience, Inc. MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit claims substantial equivalence to the Diagnostics Hybrid, Inc. ELVIS® HSV ID/Typing Test System (K971662). Table 1 identifies the characteristics of EraGen Bioscience, Inc. MultiCode® -RTx Herpes Simplex Virus 1 & 2 Kit (New Device) and the Diagnostics Hybrid, Inc. ELVIS® HSV ID/Typing Test System (Predicate Device).
| Device
Characteristic | EraGen Biosciences
MultiCode®-RTx Herpes
Simplex Virus 1 & 2 Kit
(New Device) | Diagnostics Hybrid, Inc.
ELVIS® HSV ID/Typing Test
System
(Predicate Device) |
|----------------------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Indications for Use | The MultiCode®-RTx HSV 1&2
Kit is a polymerase chain reaction
(PCR)-based qualitative in vitro
diagnostic test for the detection
and typing of herpes simplex virus
(HSV1&2) DNA in vaginal
lesions. It is indicated for use in the
detection and typing of HSV-1 or
HSV-2 in vaginal lesion swab
specimens from symptomatic
female patients as an aid in the
diagnosis of genital herpes
infection.
Warning: The device is not FDA
cleared for the use with cerebral
spinal fluid (CSF) or any lesions
other than vaginal. This assay is
not intended to be used for male
penile specimens, for prenatal
screening, or for females under
the age of 18 years. | The ELVIS® HSV ID Typing Test
System is a qualitative test
indicated for use in isolation and
identification of HSV from lesions
and body fluids suspected of
containing viable HSV-1 and/or
HSV-2. Both serotypes have been
isolated in various parts of the
body, particularly when HSV-
associated disease is indicated.
Performance of this assay has not
been established for use with
antiviral therapy or prenatal
monitoring. |
| Identification and
Typing of HSV-1
and HSV-2 | Yes. | Yes. |
| Assay Results | Qualitative. | Qualitative. |
| Assay Type | Real-Time PCR. | Cell Culture using an enzyme
linked virus inducible system. |
| Packaging | The product is supplied in labeled,
sterile tubes. The outer container
is a labeled box. | The product is supplied in a
labeled, sterile tube. The outer
container is a labeled box. |
| Analysis Software
Provided | Yes. | No. |
Table 1. Comparison of New Device with Predicate Device
5
EraGen Biosciences, Inc. - 510(k) Premarket Notification MultiCode -RTx Herpes Simplex Virus 1 & 2 Kit
| Device
Characteristic | EraGen Biosciences
MultiCode®-RTx Herpes
Simplex Virus 1 & 2 Kit
(New Device) | Diagnostics Hybrid, Inc.
ELVIS® HSV ID/Typing Test
System
(Predicate Device) |
|------------------------------------|----------------------------------------------------------------------------------------|---------------------------------------------------------------------------------------|
| Printed Results
Report Provided | Yes. | No. |
| Kit Reagent
Storage Conditions | -15°C to -30°C | 2°C to 8°C and 22°C to 28°C |
8.0 Test Principle
Following nucleic acid extraction from a patient swab specimen, a fluorophorelabeled PCR primer pair amplifies a segment of the glycoprotein B gene of HSV-I and HSV-2. The sample processing control (SPC) will also be amplified by a distinct fluorophore-labeled PCR primer pair unless there are sample processing errors, inhibitory substances in the PCR reaction, reagent failure, or instrument malfunction.
The MultiCode®-RTx system is based on an expanded genetic alphabet technology, consisting of 2'-deoxy-5-methyl-isocytidine (iC) and 2'-deoxyisoguanosine (iG) nucleotide bases also known as isobases. The isobases pair specifically with each other and not with natural nucleotides. In addition isobases are efficiently incorporated during PCR. The isobase pair allows site-specific incorporation of a dabcyl quencher directly adjacent to a fluorophore-labeled primer. During PCR amplification, a quencher-modified iGTP is incorporated by the polymerase opposite an iC and a fluorophore reporter attached to a PCR primer. If target is present and is amplified, assay fluorescence decreases with every cycle as amplification product accumulates. The decrease in assay fluorescence is monitored in real time using the Roche LightCycler 1.2 instrument. Following PCR, the amplification products are thermally denatured and assay fluorescence is monitored. The strands of the amplification products are separated and assay fluorescence increases, thus determining the melting temperature (Tm) profile of the amplicon. The sequences between the PCR primer binding sites of the HSV-1 and HSV-2 amplicons have different base compositions that are distinguished by their different melting temperatures.
9.0 Performance Characteristics
- 】. Analytical Performance
- Precision/Reproducibility: a.
The Precision/Reproducibility of the MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit was evaluated at three U.S. clinical laboratories. A panel was prepared containing six simulated HSV-1 and HSV-2 samples that included High Negative. Low Positive (near the assay limit of detection) and High Positive samples. The
6
panel, along with external HSV-1 and HSV-2 positive and negative controls, was assayed in triplicate. Kit positive and negative controls were included with each assay run. Panels and controls were tested at each site by 2 operators, 1 time each per day for 5 days (N = 900).
| | Site #1 | | | Site #2 | | | Site #3 | | | Total agreement
with
expected
results (%) | 95%
Confidence
Interval | | | |
|------------------------------------------------------|---------------------------------------------------------------------|---------|--------|----------------------|---------------------------------------------------------------------|---------|---------|----------------------|---------------------------------------------------------------------|----------------------------------------------------|-------------------------------|----------------------|------------------|------------|
| Targets | Agreement
with
expected
results-
correct /
#tested | Avg Tm¹ | %CV-Tm | Avg %
Deflection² | Agreement
with
expected
results-
correct /
#tested | Avg Tm¹ | %CV-Tm | Avg %
Deflection² | Agreement
with
expected
results-
correct /
#tested | Avg Tm¹ | %CV-Tm | Avg %
Deflection² | | |
| HS V-1 Positive Control | 10/10 | 84.3 | 0.19 | 85.0 | 10/10 | 84.7 | 0.36 | 78.3 | 10/10 | 84.7 | 0.22 | 72.8 | 30/30
(100%) | 88.4%-100% |
| HS V-2 Positive Control | 10/10 | 87.2 | 0.21 | 70.9 | 10/10 | 87.7 | 0.32 | 70.3 | 10/10 | 87.7 | 0.17 | 68.8 | 30/30
(100%) | 88.4%-100% |
| HS V-1/HS V-2 Negative
Control¹ | 10/10 | 77.9 | 0.18 | 1.5 | 10/10 | 78.0 | 0.28 | 1.4 | 10/10 | 78.0 | 0.21 | 1.8 | 30/30
(100%) | 88.4%-100% |
| PN 1750 HSV-1 Positive
External Control | 30/30 | 84.5 | 0.37 | 79.2 | 29/30 | 84.6 | 0.27 | 68.3 | 30/30 | 84.7 | 0.24 | 67.3 | 89/90
(98.9%) | 93.9%-100% |
| PN 1751 HSV-2 Positive
External Control | 30/30 | 87.4 | 0.40 | 73.0 | 29/30 | 87.6 | 0.28 | 68.2 | 30/30 | 87.7 | 0.23 | 69.3 | 89/90
(98.9%) | 93.9%-100% |
| PN 1754 HSV-1/HSV-2
Negative External
Control¹ | 30/30 | 77.6 | 0.40 | 1.6 | 30/30 | 77.7 | 0.43 | 1.3 | 30/30 | 77.8 | 0.29 | 1.1 | 90/90
(100%) | 95.9%-100% |
| HSV-1 High Negative¹ | 30/30 | 77.6 | 0.39 | 2.2 | 30/30 | 77.7 | 0.44 | 2.3 | 30/30 | 77.8 | 0.32 | 1.4 | 90/90
(100%) | 95.9%-100% |
| HSV-1 -Low Positive | 30/30 | 84.4 | 0.37 | 75.7 | 29/30 | 84.5 | 0.26 | 64.3 | 30/30 | 84.6 | 0.21 | 65.1 | 89/90
(98.9%) | 93.9%-100% |
| HSV-1 High Positive | 30/30 | 84.5 | 0.36 | 94.4 | 30/30 | 84.6 | 0.20 | 92.8 | 30/30 | 84.8 | 0.21 | 94.3 | 90/90
(100%) | 95.9%-100% |
| HSV-2 High Negative¹ | 30/30 | 77.6 | 0.41 | 1.2 | 30/30 | 77.7 | 0.28 | 1.5 | 30/30 | 77.7 | 0.27 | 1.3 | 90/90
(100%) | 95.9%-100% |
| HSV-2 Low Positive | 30/30 | 87.3 | 0.41 | 71.9 | 30/30 | 87.5 | 0.25 | 63.5 | 30/30 | 87.6 | 0.19 | 67.5 | 90/90
(100%) | 95.9%-100% |
| HSV-2 High Positive | 30/30 | 87.4 | 0.39 | 92.9 | 30/30 | 87.5 | 0.19 | 89.4 | 30/30 | 87.7 | 0.20 | 91.9 | 90/90
(100%) | 95.9%-100% |
as used, since these targets did not generate any detectable HSV-1 or HSV-2 signal.
? % defection is the individual sample net cares and can a percent of the mass as a percent of the mainum dellection of the mell career in hat assey non. Aveage % deficeris is the average deflection for that panel member across all the 10 runs for the site.
-
Linearity/Assay Reportable Range: b.
No applicable -
Traceability, Stability, Expected Values: c.
Stability studies were performed in-house to determine the shelf life for the MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit. Product claims are as follows: -
Accelerated Stability: The product is stable for a minimum of ● 15 months when stored at an accelerated temperature of 4℃.
7
-
. Real-Time Stability: The product is stable for a minimum of 6 months when stored the recommended storage temperature of -15°C to -30°C.
Real time stability studies will continue to be performed to support the shelf life of this product. -
Limit of Detection /Limit of Blank: d.
A Limit of Detection (LoD) and Limit of Blank (LoB) study was performed at EraGen Biosciences to determine the analytical LoD and LoB performance using quantified (TCID50/mL) cultures of HSV-1 and HSV-2 serially diluted in Copan Universal Transport Media. Each viral strain was extracted using the MagNA Pure LC and tested in replicates of 60 per concentration of virus. The LoD was determined to be the lowest concentration of target that was detected in at least 95% of replicates. The LoB was determined to be the highest concentration of target that was detected in ≤ 5% of replicates.
The LoD for HSV-1 was determined to be 2.0 x 103 TCID50/mL. At this concentration, 100% of samples were detected with a 95% Confidence Interval of 93.94 - 100%. The LoD for HSV-2 was determined to be 6.4 x 10 TCID50/mL. At this concentration. 98.3% of samples were detected with a 95% Confidence Interval of 91.06 – 99.96%.
The LoB for HSV-1 was determined to be 2.5 x 102 TCID50/mL. At this concentration, 3.33% of samples were detected with a 95% Confidence Interval of 0.41 – 11.53%. The LoB for HSV-2 was determined to be 4.0 TCID50/mL. At this concentration, 0.00% of samples were detected with a 95% Confidence Interval of 0.00 -5.96%.
8
| TCID50/mL | Pos/Total Calls | Positivity | 95% C.I. | |
|---|---|---|---|---|
| 4.00 x 103 | 59/59* | 100.00% | 91.06 ± 99.96% | |
| 2.00 x 103 | 59/59* | 100.00% | 91.06 ± 99.96% | LoD |
| 1.00 x 103 | 17/60 | 28.33% | 17.45 ± 41.44% | |
| 5.00 x 102 | 6/59* | 10.17% | 4.82 ± 22.57% | |
| 2.50 x 102 | 2/60 | 3.33% | 0.41 ± 11.53% | LoB |
| 1.25 x 102 | 0/60 | 0.00% | 0.00 ± 5.96% |
LoD for HSV-1
*One "Invalid" call due to baselines of amp curves by newest V2.0 RC1 software
LoD for HSV-2
| TCID50/mL | Pos/Total Calls | Positivity | 95% C.I. |
|---|---|---|---|
| 1.28 x 102 | 60/60 | 100.00% | 94.04 ± 100.00% |
| 6.40 x 101 | 59/60 | 98.33% | 91.06 ± 99.96% |
| 3.20 x 101 | 56/60 | 93.33% | 83.80 ± 98.15% |
| 1.60 x 101 | 17/57* | 29.82% | 18.43 ± 43.40% |
| 8.00 | 4/60 | 6.67% | 1.85 ± 16.20% |
| 4.00 | 0/60 | 0.00% | 0.00 ± 5.96% |
*Three "Invalid" calls due to baselines of amp curves by newest V2.0 RC1 software
Analytical Specificity and Cross Reactivity: e.
Analytical Specificity/Cross Reactivity of the MultiCode® -RTx HSV 1&2 Kit was evaluated at EraGen Biosciences. A panel was prepared containing 22 different organisms representing nearneighbors to the HSV-1 and HSV-2 virus and organisms reasonably expected to be present in vaginal swab specimens.
The cross-reactivity panel was tested in a background of Copan Universal Transport Media at the concentration indicated in the table below. Samples were extracted using the Roche MagNA Pure LC and tested in triplicate. No HSV positive results were observed for any of the organisms tested and the DNA SPC was detected in all cases. The organism panel was also spiked into HSV-1 and HSV-2 near the device's Limit of Detection (LoD) and tested.
No interference was observed from any of the tested organisms, and all the results were positive for HSV-1 or HSV-2 as expected.
9
| Organism | Concentration |
|---|---|
| Candida albicans | 2.8 x107 CFU/mL |
| Chlamydia trachomatis | 2.5 x108 EBs/mL |
| Escherichia coli | 2.0 x107 CFU/mL |
| Mycoplasma hominis | 4.5 x105 CFU/mL |
| Neisseria gonorrhoeae | 1.8 x106 CFU/mL |
| Staphylococcus aureus | 2.4 x107 CFU/mL |
| Staphylococcus saprophyticus | 1.2 x107 CFU/mL |
| Streptococcus pyogenes | 1.4 x107 CFU/mL |
| Trichomonas vaginalis | 1.1 x107 CFU/mL |
| Bacteroides fragilis | 3.3 x107 CFU/mL |
| Gardnerella vaginalis | 1.4 x106 CFU/mL |
| Mobiluncus mulieris | 1.0 x107 CFU/mL |
| Toxoplasma gondii | 6.6 x105 tachyzoites/mL |
| Treponema pallidum | 1.0 x107 CFU/mL |
| Cytomegalovirus (AD169 strain) | 4.2 x103 TCID50/mL |
| Enterovirus (Type 71) | 1.4 x104 TCID50/mL |
| Epstein-Barr virus (B95-8 strain) | 9.3 x107 copies/mL |
| Varicella Zoster virus | 2.4 x107 copies/mL |
| Human Herpes 6 virus (Z29 strain) | 1.9 x106 TCID50/mL |
| Human Herpes 7 virus (SB strain) | 3.4 x106 TCID50/mL |
| Human Papilloma virus | 5-8 x105 copies/mL |
| Rubella virus | 1.7 x104 TCID50/mL |
f. Interfering Substances:
An Interfering Substance study was performed at EraGen Biosciences to evaluate the effects of potential interfering substances on the MultiCode®-RTx HSV 1&2 Kit. A panel was prepared containing 6 substances that could reasonably be expected to be present in vaginal swab specimens. The substance panel was tested near the device's Limit of Detection (LoD) for HSV-1 and HSV-2. No interference was observed in the presence of exogenous and endogenous substances in an extractable sample.
| Substance | Concentration |
|---|---|
| Whole Blood (with EDTA) | 10% |
| Whole Blood (with EDTA) | 1% |
| Female Urine | 10% |
| Female Urine | 1% |
| Protein (Albumin) | 10 mg/ml |
10
| Substance | Concentration |
|---|---|
| Protein (Albumin) | 1 mg/ml |
| Protein (Casein) | 10 mg/ml |
| Protein (Casein) | 1 mg/ml |
| K-Y Brand Jelly | 5% |
| K-Y Brand Jelly | 0.5% |
| Acyclovir (Acycloguanosine) | 2.5 mg/ml |
| Acyclovir (Acycloguanosine) | 0.25 mg/ml |
Competitive Inhibition: ದ್ದ.
Competitive Inhibition of the MultiCode®-RTx HSV 1&2 Kit was evaluated internally using simulated samples with varying concentrations of HSV-1 virus (1X LoD to 1000X LoD) and HSV-2 virus (1X LoD to 1000X LoD). Competitive inhibition was observed. The highest concentration of co-infecting target that can be present while maintaining 95% detection of the 3X LoD target for HSV-1 was 1X LoD for HSV-2. The highest concentration of co-infecting target that can be present while maintaining 95% detection of the 3X LoD target for HSV-2 was 1X LoD for HSV-1.
Competitive Inhibition was also evaluated internally using simulated samples with equal concentrations of HSV-1 virus and HSV-2 virus (5X LoD to 500X LoD). Competitive inhibition was not observed at any of the concentrations tested when the concentrations of HSV-1 and HSV-2 were equal.
-
h. Carry-over/Contamination:
Carry-over/Contamination studies were done only with HSV-1 target, since both HSV-1 and HSV-2 share a single set of primers. The MultiCode®-RTx HSV 1&2 Kit was evaluated internally using simulated samples at the LoB and High Positive HSV-1 (1700X LoD) samples. Ten sets of samples in a sequence pattern of High Positive-LoB-LoB were aliquoted into a 32-well sample plate and extracted using the Roche MagNA Pure LC instrument. The high positive sample was positive for HSV-1 in all cases and all LoB samples were negative for HSV-1 indicating that there was no carry-over/cross-contamination of HSV-1 in the LoB samples. -
i. Assay Cut-Off:
Not Applicable
11
2. Clinical Performance:
Performance characteristics of the MultiCode®-RTx HSV 1&2 Kit were established during a prospective study performed at three U.S. clinical laboratories during 2008-2009. Vaginal lesion swab specimens were sent to the clinical laboratory for routine Herpes Simplex Virus testing.
The reference method was a cell culture based ELVIS® Herpes Simplex Virus identification and typing test. A total of 1041 vaginal swab specimens were tested by the MultiCode -RTx HSV 1&2 Kit and by the reference method.
| Herpes Simplex Virus Type 1 Comparison Results | ||||
|---|---|---|---|---|
| Reference Method | ||||
| Positive | Negative | Total | ||
| MultiCode®-RTx | ||||
| HSV 1&2 Kit | Positive | 97 | 16a | 113 |
| Negative | 8b | 920 | 928 | |
| Total | 105 | 936 | 1041 | |
| Value | 95% Confidence Interval | |||
| Sensitivity | 92.4% | 85.7 - 96.1% | ||
| Specificity | 98.3% | 97.2 - 98.9% |
Results from Prospective Study:
8 Sequence analysis detected HSV-1 in 12 of the 16 discordant samples identified as HSV-1 by MultiCode -RTx. Sequence analysis did not detect HSV-1 in 4 of the discordant samples.
b Sequence analysis detected HSV-1 in 1 of the 8 discordant samples identified as HSV-1 negative by MultiCode -RTx. Sequence analysis did not detect HSV-1 in 7 of the discordant samples. Of these 7 discordant samples: 4 of the samples were identified as HSV-2 by both MultiCode®-RTx and sequencing, 2 of the samples were negative by MultiCode -RTx and not detected by sequencing, and 1 sample was negative by MultiCode®-RTx and HSV-2 positive by sequencing.
12
| Herpes Simplex Virus Type 2 Comparison Results | ||||
|---|---|---|---|---|
| Reference Method | ||||
| Positive | Negative | Total | ||
| MultiCode®-RTx | ||||
| HSV 1&2 Kit | Positive | 198 | 53a | 251 |
| Negative | 10b | 780 | 790 | |
| Total | 208 | 833 | 1041 | |
| Value | 95% Confidence Interval | |||
| Sensitivity | 95.2% | 91.4 – 97.4% | ||
| Specificity | 93.6% | 91.8 – 95.1% |
4 Sequence analysis detected HSV-2 in 43 of the 53 discordant samples identified as HSV-2 by MultiCode -RTx. Sequence analysis did not detect HSV-2 in 10 of the discordant samples.
P Sequence analysis detected HSV-2 in 2 of the 10 discordant samples identified as HSV-2 negative by MultiCode - RTx. Sequence analysis did not detect HSV-2 in 8 of the discordant samples. These 8 samples were identified as HSV-1 by both MultiCode® RTx and sequencing.
A total of 69 specimens were reference method negative and MultiCode -RTx HSV 1&2 Kit positive for HSV-1 or HSV-2. DNA sequencing analysis agreed in 55 of these 69 specimens with the MultiCode -RTx HSV 1&2 Kit.
10.0 Statement of Supporting Data
Real time stability studies will be ongoing to support the shelf life of this product. Supporting data for all studies performed is retained on file at EraGen Biosciences, Inc.
13
Image /page/13/Picture/0 description: The image shows the seal of the Department of Health & Human Services - USA. The seal is circular and contains the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. In the center of the seal is a stylized image of three human figures.
DEPARTMENT OF HEALTH & HUMAN SERVICES
Public Health Service
Food and Drug Administration 10903 New Hampshire Avenue Document Mail Center-WO66-G609 Silver Spring, MD 20993-0002
MAY 1 2 2010
Mr. Randal Vader Vice President Quality, Regulatory and Clinical Affairs Eragen Biosciences, Inc. 918 Deming Way Suite 201 Madison, Wisconsin 53717
Re: K100336
| Trade/Device Name: | MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit |
|---|---|
| Regulation Number: | 21CFR §866.3305 |
| Regulatory Name: | Herpes Simplex Virus Nucleic Acid Amplification Assay |
| Regulatory Class: | Class II |
| Product Code: | OQO |
| Dated: | March 6, 2010 |
| Received: | March 8, 2010 |
Dear Mr. Vader:
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.
If 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 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 of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements
14
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. 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 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 Part 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/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 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,
Vally, actopse
Sally A. Hojvat, M.Sc., Ph.D Director Division of Microbiology Devices Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health
Enclosure
15
Indications for Use
510(k) Number (if known): K100336
Device Name: MultiCode®-RTx Herpes Simplex Virus 1 & 2 Kit
Indications for Use:
The MultiCode®-RTx HSV 1&2 Kit is a polymerase chain reaction (PCR)-based qualitative in vitro diagnostic test for the detection and typing of herpes simplex virus (HSV1&2) DNA in vaginal lesions. It is indicated for use in the detection and typing of HSV-1 or HSV-2 in vaginal lesion swab specimens from symptomatic female patients as an aid in the diagnosis of genital herpes infection.
Warning: The device is not FDA cleared for the use with cerebral spinal fluid (CSF) or any lesions other than vaginal. This assay is not intended to be used for male penile specimens, for prenatal screening, or for females under the age of 18 years.
| Prescription Use
| (Part 21 CFR 801 Subpart D) | X AN |
|---|---|
| ------------------------------------------------- | ------ |
ND/OR
Over-The-Counter Use (21 CFR 801 Subpart C)
(PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE OF NEEDED)
Concurrence of CDRH, Office of Device Evaluation (ODE)
| Division Sign-Off | |
|---|---|
| ------------------- | -- |
Office of In Vitro Diagnostic Device Evaluation and Safety
| 510(k) | k100 336 |
|---|---|
| Page | 1 of 1 |