(220 days)
Not Found
No
The device is a sample collection and transport system, not a diagnostic or analytical device that would typically incorporate AI/ML for data processing or interpretation. The summary focuses on the physical components, media composition, and performance related to sample stability and inactivation. There is no mention of AI, ML, or related concepts.
No.
The device is intended for the collection, inactivation, stabilization, and transportation of samples for molecular detection, not for treating a disease or condition.
No
Explanation: The device is intended for the collection, inactivation, stabilization, and transportation of samples for molecular detection in a laboratory. It is not designed to perform a diagnostic test itself or provide an interpretation of results.
No
The device description clearly states it is a physical system consisting of a polypropylene tube, transport media, and swabs. It is a hardware device for specimen collection and transport, not a software-only device.
Based on the provided information, the NEST ITM device is an In Vitro Diagnostic (IVD).
Here's why:
- Intended Use: The intended use explicitly states that the system is for the "collection, inactivation stabilization and transportation of pharyngeal and nasal swabs suspected of containing adenovirus, influenza A virus or parainfluenza virus 2 from the collection site to the testing laboratory." It also states that the "specimen transported in NEST ITM can be used for molecular detection in the laboratory." This clearly indicates that the device is intended for use in the examination of specimens derived from the human body to provide information for diagnostic purposes.
- Device Description: The description details a preservation tube and transport media designed to handle human samples suspected of containing specific viruses for laboratory testing.
- Performance Studies: The document includes performance data related to detection limit, viral stability, and inactivation, which are all relevant to the performance of an IVD device used for specimen handling prior to diagnostic testing.
- Predicate Device: The mention of a predicate device (PrimeStore MTM, K number DEN170029) strongly suggests that this device is being compared to another device already classified as an IVD.
Therefore, the NEST ITM fits the definition of an In Vitro Diagnostic device as it is intended for the collection and preparation of human specimens for subsequent diagnostic testing in a laboratory setting.
N/A
Intended Use / Indications for Use
NEST ITM is an enclosed system intended for the collection, inactivation stabilization and transportation of pharyngeal and nasal swabs suspected of containing adenovirus, influenza A virus or parainfluenza virus 2 from the collection site to the testing laboratory. The specimen transported in NEST ITM can be used for molecular detection in the laboratory.
Product codes (comma separated list FDA assigned to the subject device)
QBD
Device Description
The NEST ITM is a medical-grade Polypropylene preservation tube (5 mL and 10 mL) filled with 2.5 mL ITM Inactivated Transport Media for 5 mL tube or 3 mL Inactive Transport Media for 10 mL tube, with or without the sterile swabs. NEST ITM is composed of Guanidine isothiocyanate, TCEP, sodium acetate, PEG-6000, Tris, Hcl, purified water in order to inactivate infectious unprocessed oropharyngeal and nasopharyngeal samples which are suspected of containing adenovirus, influenza A virus or parainfluenza virus 2 from human samples. For both oropharyngeal and nasopharyngeal swabs, the swab head is made of flocked nylon fiber, and the rod is made of ABS (acrylonitrile butadiene styrene).
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
pharyngeal and nasal
Indicated Patient Age Range
Not Found
Intended User / Care Setting
collection site to the testing laboratory
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
Not Found
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Non-Clinical Performance Data: Wuxi Nest completed non-clinical tests to demonstrate safety and effectiveness and substantial equivalence to the predicate device. Results confirmed that design inputs and performance specifications are met.
- FDA Recognized Consensus Standards:
- ASTM F1980-2016, Standard Guide for Accelerated Aging of Sterile Barrier Systems for Medical Devices - Passed
- ASTM D4169-2016, Standard practice for performance testing of shipping containers and systems - Passed
- ISO 11607-1:2019, Packaging for terminally sterilized medical devices -- Part 1: Requirements for materials, sterile barrier systems and packaging systems - Passed
- ISO 11607-2:2019, Packaging for terminally sterilized medical devices-Part 2: Validation requirements for forming, sealing and assembly processes - Passed
- ISO 15223-1:2016, Medical devices-Symbols to be used with medical device labels, labelling and information to be supplied-Part 1: General requirement - Full Compliance with.
- ISO 14971:2019, Medical devices-Application of risk management to medical devices - Full Compliance with.
- Non-FDA Recognized Consensus Standard:
- CLSI MM13-A (Replaces MM13-P) Collection, Transport, Preparation, and Storage of Specimens for Molecular Methods; Approved Guideline - Full Compliance with.
- Shelf life:
- The shelf life for the NEST ITM is 12 months after the date of manufacture.
- Stability was performed using Realtime and Accelerated stability on three (3) lots.
- Stability looked for bacterial and fungal growth, media properties (appearance, pH), and viral stabilization at room temperature for 15 days, demonstrating nucleic acid stability.
- Sterilization:
- The DNA/RNA Shield Collection tube with media are not sold as sterile nor are they intended to be sterilized by the user.
- These vials are single use devices that do not require cleaning by the operator.
- The Swabs are individually packaged and sold as sterile.
- Detection Limit:
- LoD testing was conducted to determine the lowest concentration of analyte with a >95% detection rate.
- LoD studies for Adenovirus, Influenza A, and Parainfluenza virus 2 were designed using validated assays.
- Preliminary LoD testing: Adenovirus, Influenza A, and Parainfluenza 2 virus were spiked at 1.0x10^4, 5.0x10^3, and 1.0x10^2 copies/mL into NEST ITM with a swab. A validated PCR assay determined the LoD to be 5.0x10^2 for each virus.
- Confirmatory LoD testing: Performed at 5.0x10^2 copies/mL with 20 replicates. All 20 replicates had recoverable concentrations and met the predefined acceptance criteria.
- Viral Stability:
- Stability of Adenovirus, Influenza A, and Parainfluenza 2 virus at 1 x LoD (5.0x10^2 copies/mL) was evaluated by spiking virus into simulated matrix incubated in NEST ITM at refrigerated temperature (4°C) for 15 days and ambient temperature (25°C) for 15 days.
- Testing used at least 20 replicates for each virus, time point, and storage condition, across three lots.
- The validated PCR assays were run with all applicable controls.
- Pre-defined acceptance criteria: (+/-) 3.0 Ct from the initial time zero value.
- Results: Maximum average variation of 0.8 Ct over 15 days at 25°C and 0.9 Ct over 15 days at 2-8°C.
- Inactivation:
- Adenovirus, Influenza A, and Parainfluenza 2 virus at 1.0 x 10^7 TCID50/ml was incubated with NEST ITM for 10 seconds.
- The NEST ITM rapidly inactivated all viruses tested with a >4.0 log reduction at a 1:10 specimen to media concentration at 10 seconds.
- Viral CPE could not be observed at
§ 866.2950 Microbial nucleic acid storage and stabilization device.
(a)
Identification. A microbial nucleic acid storage and stabilization device is a device that consists of a container and reagents intended to stabilize microbial nucleic acids in human specimens for subsequent isolation and purification of nucleic acids for further molecular testing. The device is not intended for preserving morphology or viability of microorganisms.(b)
Classification. Class II (special controls). The special controls for this device are:(1) The intended use for the labeling required under § 809.10 of this chapter must include a detailed description of microorganisms and types of human specimens intended to be preserved.
(2) The labeling required under § 809.10(b) of this chapter must include the following:
(i) A detailed device description, including all device components;
(ii) Performance characteristics from applicable analytical studies, including nucleic acid stability and microorganism inactivation;
(iii) A limiting statement that erroneous results may occur when the transport device is not compatible with molecular testing; and
(iv) A limiting statement that the device has only been validated to preserve the representative microorganisms used in the analytical studies.
(3) Design verification and validation must include the following:
(i) Overall device design, including all device components and all control elements incorporated into the analytical validation procedures;
(ii) Thorough description of the microorganisms and methodology used in the validation of the device including, extraction platforms and assays used for the detection of preserved nucleic acids; and
(iii) The limit of detection (LoD) of the molecular test used to establish microorganism nucleic acid stability.
0
Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo in blue, with the words "U.S. FOOD & DRUG" stacked on top of "ADMINISTRATION".
Wuxi Nest Biotechnology Co., Ltd. % Giselle Zhang Regulatory Consultant Emergo Global Consulting, LLC 2500 Bee Cave Road 1 Suite 300 Austin, Texas 78746
September 20, 2021
Re: K210440
Trade/Device Name: Disposable Sampler Inactivated Transport Media, Nest ITM Regulation Number: 21 CFR 866.2950 Regulation Name: Microbial Nucleic Acid Storage And Stabilization Device Regulatory Class: Class II Product Code: OBD Dated: February 10, 2021 Received: February 12, 2021
Dear Giselle Zhang:
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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
1
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
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 https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely.
Kristian Roth, Ph.D. Branch Chief Bacterial Respiratory and Medical Counter Measures Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Ouality Center for Devices and Radiological Health
2
Indications for Use
Form Approved: OMB No. 0910-0120 Expiration Date: 06/30/2023 See PRA Statement below.
510(k) Number (if known)
Device Name
Disposable Sampler Inactivated Transport Media
Indications for Use (Describe)
NEST ITM is an enclosed system intended for the collection, inactivation stabilization and transportation of pharyngeal and nasal swabs suspected of containing adenovirus, influenza A virus or parainfluenza virus 2 from the collection site to the testing laboratory. The specimen transported in NEST ITM can be used for molecular detection in the laboratory.
| Type of Use (Select one or both, as applicable) | |
|---|---|
| Prescription Use (Part 21 CFR 801 Subpart D) | Over-The-Counter Use (21 CFR 801 Subpart C) |
CONTINUE ON A SEPARATE PAGE IF NEEDED.
This section applies only to requirements of the Paperwork Reduction Act of 1995.
DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.
The burden time for this collection of information is estimated to average 79 hours per response, including the
time to review instructions, search existing data sources, gather and maintain the data needed and complete
and review the collection of information. Send comments regarding this burden estimate or any other aspect
of this information collection, including suggestions for reducing this burden, to:
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"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
5. 510(k) Summary
The following information is provided in accordance with 21 CFR 807.92 for the Premarket 510(k) Summary:
5.1 Submitter Information
| Company: | Cheng Zhiwei
RA Manager
Wuxi NEST Biotechnology Co., Ltd.
No.530 Xida Road, New District
Wuxi, Jiangsu 214012 China
Telephone: 86+510-68006788
Fax: N/A
project01@nest-wuxi.com |
|------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Contact: | Giselle Zhang
Regulatory Consultant
Emergo Global Consulting, LLC
2500 Bee Cave Road, Building 1, Suite 300
Austin, Texas 78746 USA
Telephone: (512) 327-9997
Fax: (512) 327-9998
LST.AUS.ProjectManagement@ul.com |
| Date Summary Prepared: | May 7, 2021 |
5.2 Name of the Device
| Trade Name: | Disposable Sampler Inactivated Transport Media |
|---|---|
| Common Name: | Microbial nucleic acid storage and stabilization device |
| Classification Name: | Microbiology |
| Review Panel: | Microbiology (MI) |
| Regulation: | 866.2950 |
| Class: | Class II |
| Product Code: | QBD |
5.3 Equivalence Claimed to Predicate Device
The Disposable Sampler Inactivated Transport Media is equivalent to the PrimeStore MTM (DEN170029), manufactured by Longhorn Vaccines and Diagnostics, LLC.
4
5.4 Device Description
The NEST ITM is a medical-grade Polypropylene preservation tube (5 mL and 10 mL) filled with 2.5 mL ITM Inactivated Transport Media for 5 mL tube or 3 mL Inactive Transport Media for 10 mL tube, with or without the sterile swabs. NEST ITM is composed of Guanidine isothiocyanate, TCEP, sodium acetate, PEG-6000, Tris, Hcl, purified water in order to inactivate infectious unprocessed oropharyngeal and nasopharyngeal samples which are suspected of containing adenovirus, influenza A virus or parainfluenza virus 2 from human samples. For both oropharyngeal and nasopharyngeal swabs, the swab head is made of flocked nylon fiber, and the rod is made of ABS (acrylonitrile butadiene styrene).
5.5 Indication for Use Statement
NEST ITM is an enclosed system intended for the collection stabilization and transportation of pharyngeal and nasal swabs suspected of containing adenovirus, influenza A virus or parainfluenza virus 2 from the collection site to the testing laboratory. The specimen transported in NEST ITM can be used for molecular detection in the laboratory.
5.6 Substantial Equivalence Discussion
The following table compares the Disposable Sampler Inactivated Transport Media to the predicate device with respect to indications for use, principles of operation, technological characteristics, materials, and performance, and forms the basis for the determination of substantial equivalence. The subject device does not raise any new questions of safety or effectiveness as compared to the predicate device.
| Device & Predicate Device(s): | K210440 | DEN170029 |
|---|---|---|
| Device Trade Name | Nest ITM | PrimeStore MTM |
| General Device Characteristic | ||
| Similarities | K210440 | DEN170029 |
| Intended Use/Indications for | ||
| Use | NEST ITM is an enclosed system | |
| intended for the collection, | ||
| inactivation stabilization and | ||
| transportation of pharyngeal | ||
| and nasal swabs suspected of | ||
| containing adenovirus, influenza | ||
| A virus or parainfluenza virus 2 | ||
| from the collection site to the | ||
| testing laboratory. The | ||
| specimen transported in NEST | ||
| ITM can be used for molecular | ||
| detection in the laboratory. | PrimeStore MTM is intended for | |
| the stabilization, transportation | ||
| and inactivation of infectious | ||
| unprocessed nasal washes | ||
| suspected of containing | ||
| Influenza A virus RNA. | ||
| PrimeStore MTM is also | ||
| intended for the stabilization, | ||
| transportation and inactivation | ||
| of infectious unprocessed | ||
| sputum samples suspected of | ||
| containing Mycobacterium |
5
| | | tuberculosis DNA from human
samples. |
|----------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------|
| Inactivation tested | >4.0 log reduction in
concentration at 10 seconds | Same |
| Storage temperatures | 2-8 ºC up to 25ºC | Same |
| General Device Characteristic
Differences | K210440 | DEN170029 |
| Specimen stability | NEST ITM preserves Adenovirus,
Influenza A virus or
Parainfluenza for 15 days at 2-8
ºC and 25ºC | Primestore MTM medium
preserves influenza A RNA for
up to 8 days at 27°C and 29 days
at 4°C |
| Specimen Type | Nasopharyngeal or
Oropharyngeal Swab | Nasal washes and sputum
samples |
| Analyte | Nasopharyngeal or
Oropharyngeal swab suspected
of containing Adenovirus,
Influenza A virus or
Parainfluenza virus 2. | Nasal wash suspected of
containing Influenza A virus.
Sputum samples suspected of
containing MTB. |
5.7 Non-Clinical Performance Data
To demonstrate safety and effectiveness of Disposable Sampler Inactivated Transport Media and to show substantial equivalence to the predicate device, Wuxi Nest completed the following non-clinical tests. Results confirm that the design inputs and performance specifications for the device are met. The Disposable Sampler Inactivated Transport Media passed the testing in accordance with internal requirements, national standards, and international standards shown below, supporting its safety and effectiveness, and its substantial equivalence to the predicate device:
-
- FDA Recognized Consensus Standards:
- ASTM F1980-2016, Standard Guide for Accelerated Aging of Sterile Barrier Systems for Medical Devices -Passed
- . ASTM D4169-2016, Standard practice for performance testing of shipping containers and systems - Passed
- ISO 11607-1:2019, Packaging for terminally sterilized medical devices -- Part 1: Requirements for materials, sterile barrier systems and packaging systems - Passed
- . ISO 11607-2:2019, Packaging for terminally sterilized medical devices-Part 2: Validation requirements for forming, sealing and assembly processes - Passed
6
- . ISO 15223-1:2016, Medical devices-Symbols to be used with medical device labels, labelling and information to be supplied-Part 1: General requirement - Full Compliance with.
- . ISO 14971:2019, Medical devices-Application of risk management to medical devices - Full Compliance with.
2. Non-FDA Recognized Consensus Standard:
- . CLSI MM13-A (Replaces MM13-P) Collection, Transport, Preparation, and Storage of Specimens for Molecular Methods; Approved Guideline - Full Compliance with. Justification: The standard is not recognized by FDA, but Wuxi Nest believes that the standard is applicable to the type of device to ensure the safety and performance of the device.
3. Shelf life:
The shelf life for the NEST ITM is 12 months after the date of manufacture. The stability of the NEST ITM was performed using Realtime and Accelerated stability on a total of three (3) lots. Stability looked for bacterial and fungal growth in the media along with properties of the media, appearance, pH, and then confirmed with viral stabilization at room temperature to the claimed 15 days demonstrating the stability of nucleic acids was not diminished with the age of media.
4. Sterilization:
The DNA/RNA Shield Collection tube with media are not sold as sterile nor are they intended to be sterilized by the user. These vials are single use devices that do not require cleaning by the operator. The Swabs are individually packages and sold as sterile.
5. Detection Limit:
LoD testing was conducted to determine the lowest concentration of analyte that can be detected with a greater than 95% detection rate. The LoD studies for Adenovirus, Influenza A, and Parainfluenza virus 2 were designed using validated assays to establish a concentration of organisms used for additional testing noted below.
LoD testing was initially performed by spiking multiple concentrations of Adenovirus, Influenza A, and Parainfluenza 2 virus into contrived matrix and spiking it onto a swab. Adenovirus, Influenza A, and Parainfluenza 2 virus were spike at a final concertation range of 1.0x10*, 5.0x10', and 1.0x10². copies/mL into the NEST ITM with a swab. A validated PCR assay was use to detemine the LoD to be 5.0x104 for each of the three viruses. Table 1 below shows the results of preliminary LoD for Adenovirus, influenza A, and Parainfluenza virus 2.
| Concentration
(copies/mL) | Adenovirus, 5
Reps Average
(Ct) | SD
(Ct) | Influenza A, 5
Rep Average
(Ct) | SD
(Ct) | Parainfluenza
virus 2, 5 Rep
Average (Ct) | SD
(Ct) |
|------------------------------|---------------------------------------|------------|---------------------------------------|------------|-------------------------------------------------|------------|
| 1.0×104 | 29.3 | 0.18 | 32.04 | 1.08% | 31.07 | 1.08% |
Table 1. Preliminary Limit of Detection
7
| 1.0x103 | 32.0 | 0.26 | 34.75 | 2.11% | 34.28 | 2.18% |
|---|---|---|---|---|---|---|
| 5.0x102 | 34.3 | 0.95 | 35.48 | 1.97% | 36.89 | 2.98% |
| 1.0x102 | >40 | - | >40 | - | >40 | - |
Confirmatory LoD testing was provided at a concentration of 5.0x102 copies/mL with 20 replicates. The validated assay had an LoD with an acceptance criteria of virus detection at a concentration range 5.0x102 copies/mL. The same detection range was replicated with the NEST ITM and further determine by the concentration that yielded at least a 95% of the replicates were recoverable within this range. At a concentration of 5.0x102 copies/mL, 20 of 20 replicates had recoverable concentrations. Viral nuclic acids were extracted using a nucleic acid (DNA/RNA) extraction and purification kits (spin column) (SC903-50) (Wuxi TechstarTechnology Co., Ltd.) and amplified using the respective kits on the ABI 7500. The average Ct values for each virus are listed below in Table. 2.
| Ct Value | |||
|---|---|---|---|
| Replicates | Adenovirus | Influenza A | Parainfluenza virus 2 |
| 1 | 36.2 | 32.0 | 38.0 |
| 2 | 37.0 | 32.4 | 38.2 |
| 3 | 38.1 | 35.4 | 37.8 |
| 4 | 38.1 | 35.2 | 36.7 |
| 5 | 36.8 | 35.4 | 36.4 |
| 6 | 36.9 | 37.0 | 38.4 |
| 7 | 35.5 | 35.8 | 37.3 |
| 8 | 36.1 | 35.0 | 35.2 |
| 9 | 37.2 | 35.4 | 35.9 |
| 10 | 35.8 | 35.7 | 36.5 |
| 11 | 37.7 | 35.8 | 36.2 |
| 12 | 36.3 | 35.8 | 36.2 |
| 13 | 36.4 | 35.3 | 34.2 |
| 14 | 37.5 | 36.2 | 35.3 |
| 15 | 36.7 | 34.1 | 36.1 |
| 16 | 35.7 | 35.6 | 35.1 |
| 17 | 37.0 | 35.9 | 36.6 |
| 18 | 37.0 | 35.7 | 35.8 |
| 19 | 36.8 | 36.2 | 37.5 |
| 20 | 37.1 | 35.0 | 36.3 |
| AVG: | 36.8 | 35.2 | 36.3 |
| SD: | 0.73 | 1.2 | 1.1 |
Table 2 Average Ct Values for Each Virus
LoD testing at 5.0x102 copies/mL resulted in all 20 replicates for the concentration meeting the predefined acceptance criteria.
-
- Viral Stability
8
The stability of Adenovirus, Influenza A, and Parainfluenza 2 virus at 1 x LoD (5.0x10 copies/mL) was evalutaed by spiking virus into simulated matrix incubated in the NEST ITM at refridgerated temperature (4°C, 39°F) for 15 days (see Table 3), and ambient temperature (25°C, 77° F) for 15 days (see Table 4). Validated PCR assays was used to determine stability of Adenovirus, Influenza A, and Parainfluenza 2 virus in the NEST ITM. The stability study analyzed a total of three lots near the manufacturer claimed 12-month stability. Testing used at least 20 replicates for each virus, time point and stoage condition. An initial time point designated as Day 0 was included as the initial G average for each of the two temperature ranges tested. Testing at three time points was performed at Day 0, 9 and 15 for refrigerated temperature (2-8°C, 36-39°F), and three time points, Day 0, 9, and 15, for ambient temperature (25°C, 77°F).
The validated PCR assays were run with all applicalbe controls to valid and confirm the detection of the target virus, Adenovirus, Influenza A, and Parainfluenza virus 2. A pre-defined acceptance criteria of (+/-) 3.0 C; from the initial time zero value was the acceptance criteria.
| Day 0 | Day 9 | Day 15 | |
|---|---|---|---|
| Adenovirus AVG (Ct): | 36.0 | 36.7 | 36.9 |
| CV (Ct): | 2.15% | 1.8% | 2.0% |
| Influenza A AVG (Ct): | 35.4 | 35.4 | 35.7 |
| CV (Ct): | 2.0% | 2.3% | 2.5% |
| Parainfluenza 2 AVG (Ct): | 36.1 | 37.0 | 37.0 |
| CV (Ct): | 2.0% | 2.0% | 2.4% |
Table 3. Adenovirus, Influenza A, and Parainfluenza 2 virus (5.0x10² copies/mL) stability at 4°C
| Table 4. Adenovirus, Influenza A, and Parainfluenza 2 virus (5.0x102 copies/mL) stability 25°C | |||||
|---|---|---|---|---|---|
| ------------------------------------------------------------------------------------------------ | -- | -- | -- | -- | -- |
| Day 0 | Day 9 | Day 15 | |
|---|---|---|---|
| Adenovirus AVG (Ct): | 36.1 | 37.0 | 36.9 |
| CV (Ct): | 2.0% | 2.4% | 2.0% |
| Influenza A AVG (Ct): | 35.3 | 36.1 | 35.3 |
| CV (Ct): | 2.4% | 2.6% | 2.1% |
| Parainfluenza 2 AVG (Ct): | 36.2 | 36.9 | 37.0 |
| CV (Ct): | 2.3% | 2.6% | 2.1% |
Stability testing of RNA from whole Adenovirus, Influenza A, and Parainfluenza 2 virus were spiked into matrix and stored in NEST ITM, resulted in a maximum average variation of 0.8 G over 15 days at 25°C and a maximum variation of 0.9 Ct over 15 days at 2-8°C.
-
- Inactivation
Adenovirus, Influenza A, and Parainfluenza 2 virus at a concentration of 1.0 x 107 TClD50/ml was incubated with NEST ITM for 10 seconds. Each virus only and NEST ITM were also incubated accordingly to serve as controls. Adenovirus, Influenza A, and Parainfluenza 2 virus (virus alone), Virus and NEST ITM, or NEST ITM alone was then inculated on to cell cultures after incubation. Four days after inoculation, the cells were fixed and stained with 0.06% crystal violet in 1% glutaraldeyde. Cells that did not take up the stain were considered evidence of a viral cytopathic effect (CPE) and as
- Inactivation
9
a result were considered a measure of viral viability. The titer of the virus CPE was calcuated and recorded as the TCID50.
Inactivation time:
The NEST ITM showed no cytotoxicity on MDCK cells at a 1:1,000 dilution factor and greater; therefore at least a 1:1,000 dilution factor is needed to avoid a direct cytotoxic effect of the NEST ITM. Adenovirus, Influenza A, and Parainfluenza 2 virus were then exposed to NEST ITM for 10 seconds prior to serial 10 fold dilutions and incubations (final concentration 4.0 log reduction at a 1:10 specimen to media concentration at 10 seconds. Viral CPE could not be observed at