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.

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).

AI/ML Overview

The provided text describes the non-clinical performance data for the "Disposable Sampler Inactivated Transport Media (NEST ITM)". This device is a microbial nucleic acid storage and stabilization device, not an AI/ML-based device. Therefore, many of the requested criteria related to AI/ML device performance (e.g., human-in-the-loop, MRMC studies, ground truth establishment for deep learning models) are not applicable.

However, I can extract information related to the device's functional performance, which serves as its "acceptance criteria" for demonstrating substantial equivalence to a predicate device.

Here's the relevant information:

Device: Disposable Sampler Inactivated Transport Media (NEST ITM)
Purpose: Collection, inactivation, stabilization, and transportation of pharyngeal and nasal swabs suspected of containing adenovirus, influenza A virus, or parainfluenza virus 2 for molecular detection.

Acceptance Criteria and Reported Device Performance

The "acceptance criteria" for this device are its ability to:

Inactivate target viruses rapidly.
Preserve the nucleic acids of the target viruses over a specified period and temperatures (stability).
Detect the target viruses at a low concentration (Limit of Detection - LoD).

Table of Acceptance Criteria and Reported Device Performance:

Feature/Test	Acceptance Criteria (Internal/Implicit)	Reported Device Performance
Inactivation	>4.0 log reduction in viral concentration (similar to predicate) within 10 seconds. Absence of viral cytopathic effect (CPE) at specific dilutions.	Rapidly inactivated all tested viruses (Adenovirus, Influenza A, Parainfluenza 2 virus) with >4.0 log reduction at a 1:10 specimen to media concentration at 10 seconds. Viral CPE could not be observed at < 3.0 logs due to cellular destruction by NEST ITM.
LoD (Limit of Detection)	Virus detection at a concentration range of 5.0x10^2 copies/mL, with at least 95% of replicates recoverable.	Confirmatory LoD testing confirmed a detection range of 5.0x10^2 copies/mL for Adenovirus, Influenza A, and Parainfluenza 2 virus. 20 out of 20 replicates (100%) had recoverable concentrations at this level. Preliminary LoD also showed detection down to 5.0x10^2 copies/mL.
Viral Stability	Pre-defined acceptance criteria of (+/-) 3.0 Ct from the initial time zero value for all tested viruses (Adenovirus, Influenza A, Parainfluenza 2 virus).	Stability evaluated at 1 x LoD (5.0x10^2 copies/mL).At 4°C for 15 days: Max average variation of 0.9 Ct.At 25°C for 15 days: Max average variation of 0.8 Ct.The device preserves Adenovirus, Influenza A virus, or Parainfluenza for 15 days at 2-8 ºC and 25ºC (stated as a difference from predicate, indicating performance). All data presented is well within the +/- 3.0 Ct acceptance criteria.
Shelf Life	Demonstrate stability for 12 months post-manufacture.	Stability studies (Realtime and Accelerated) on 3 lots demonstrated stability of nucleic acids (no diminished detection with age of media) over the claimed 12-month shelf life. Tested for bacterial/fungal growth, appearance, pH, and viral stabilization.

Study Details:

Sample Size Used for Test Set and Data Provenance:
- LoD Test Set:
  - Preliminary LoD: 5 replicates per concentration for each of the 3 viruses (Adenovirus, Influenza A, Parainfluenza 2 virus) at multiple concentrations (1.0x10^4, 1.0x10^3, 5.0x10^2, 1.0x10^2 copies/mL).
  - Confirmatory LoD: 20 replicates per virus (Adenovirus, Influenza A, Parainfluenza 2 virus) at 5.0x10^2 copies/mL.
- Viral Stability Test Set: At least 20 replicates for each virus (Adenovirus, Influenza A, Parainfluenza 2 virus) per time point (Day 0, 9, 15) and storage condition (4°C, 25°C). This means a total of 20 x 3 x 2 = 120 samples per virus for stability, across 3 lots.
- Inactivation Test Set: Not explicitly stated as a number of replicates, but the method suggests controlled laboratory experiments comparing virus only, virus + NEST ITM, and NEST ITM only.
- Data Provenance: The studies were conducted by Wuxi Nest Biotechnology Co., Ltd. The data is based on laboratory testing of synthetic viral material (spiked into contrived matrix) rather than patient samples. No mention of country of origin of data beyond the manufacturer's location (China). These are non-clinical, prospective studies designed to evaluate product performance.
Number of Experts Used to Establish Ground Truth for Test Set and Qualifications of those Experts:
- Not applicable. This is a non-clinical, in-vitro diagnostic device component. The "ground truth" for these tests is the known concentration of spiked virus or the measured reduction in viral titer / preservation of nucleic acids using validated laboratory assays (e.g., PCR, TCID50). There's no human expert adjudication of images or clinical outcomes in these studies.
Adjudication Method for the Test Set:
- Not applicable. As described above, these are lab-based quantitative measurements using predefined assay thresholds and controls.
If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was done:
- No. This type of study is typically performed for AI/ML diagnostic aids for image interpretation or similar tasks that directly assist human readers. This device is a sample transport media.
If a Standalone (i.e., algorithm only without human-in-the-loop performance) was done:
- Not applicable. This is not an algorithm. The "performance" is the physical and chemical properties of the transport media.
Type of Ground Truth Used:
- For LoD and Viral Stability: The "ground truth" is the known concentration of spiked virus (e.g., 5.0x10^2 copies/mL) and subsequent detection/quantification using validated molecular methods (PCR, measured in Ct values).
- For Inactivation: The "ground truth" is the initial viral titer (TCID50) and the measured reduction in viable virus after exposure to the transport media, assessed by absence of cytopathic effect (CPE) in cell culture.
Sample Size for the Training Set:
- Not applicable. This is not an AI/ML device that requires a training set. The device formulation and manufacturing processes are developed through R&D and quality control, not machine learning.
How the Ground Truth for the Training Set Was Established:
- Not applicable.

In summary, the document details the rigorous non-clinical laboratory testing performed to demonstrate that the NEST ITM meets its functional performance requirements for viral inactivation, nucleic acid stabilization, and detection limit, thereby supporting its substantial equivalence to the predicate device. The acceptance criteria and the proof align with the nature of a microbial nucleic acid storage and stabilization device.

Summary

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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.

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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

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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)

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.

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DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

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and review the collection of information. Send comments regarding this burden estimate or any other aspect
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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."

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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 ZhiweiRA ManagerWuxi NEST Biotechnology Co., Ltd.No.530 Xida Road, New DistrictWuxi, Jiangsu 214012 ChinaTelephone: 86+510-68006788Fax: N/Aproject01@nest-wuxi.com
Contact:	Giselle ZhangRegulatory ConsultantEmergo Global Consulting, LLC2500 Bee Cave Road, Building 1, Suite 300Austin, Texas 78746 USATelephone: (512) 327-9997Fax: (512) 327-9998LST.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.

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5.4 Device Description

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 CharacteristicSimilarities	K210440	DEN170029
Intended Use/Indications forUse	NEST ITM is an enclosed systemintended for the collection,inactivation stabilization andtransportation of pharyngealand nasal swabs suspected ofcontaining adenovirus, influenzaA virus or parainfluenza virus 2from the collection site to thetesting laboratory. Thespecimen transported in NESTITM can be used for moleculardetection in the laboratory.	PrimeStore MTM is intended forthe stabilization, transportationand inactivation of infectiousunprocessed nasal washessuspected of containingInfluenza A virus RNA.PrimeStore MTM is alsointended for the stabilization,transportation and inactivationof infectious unprocessedsputum samples suspected ofcontaining Mycobacterium

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		tuberculosis DNA from humansamples.
Inactivation tested	>4.0 log reduction inconcentration at 10 seconds	Same
Storage temperatures	2-8 ºC up to 25ºC	Same
General Device CharacteristicDifferences	K210440	DEN170029
Specimen stability	NEST ITM preserves Adenovirus,Influenza A virus orParainfluenza for 15 days at 2-8ºC and 25ºC	Primestore MTM mediumpreserves influenza A RNA forup to 8 days at 27°C and 29 daysat 4°C
Specimen Type	Nasopharyngeal orOropharyngeal Swab	Nasal washes and sputumsamples
Analyte	Nasopharyngeal orOropharyngeal swab suspectedof containing Adenovirus,Influenza A virus orParainfluenza virus 2.	Nasal wash suspected ofcontaining Influenza A virus.Sputum samples suspected ofcontaining 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:

1. 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

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. 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, 5Reps Average(Ct)	SD(Ct)	Influenza A, 5Rep Average(Ct)	SD(Ct)	Parainfluenzavirus 2, 5 RepAverage (Ct)	SD(Ct)
1.0×104	29.3	0.18	32.04	1.08%	31.07	1.08%

Table 1. Preliminary Limit of Detection

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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.

1. Viral Stability

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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.

1. 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

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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 <10 TCIDso/mL), while Influenza Adenovirus, Influenza A, and Parainfluenza 2 virus only samples had viral loads of greater than 1.0 x 10° TCID50 of virus and NEST ITM alone was diluted 1:1000 to see no CPE. 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 < 3.0 logs due to cellular destruction by NEST ITM See Table 5 below.

10s incubation	Adenovirus TCID50 (log)	Influenza A TCID50 (log)	Influenza A TCID50 (log)
Virus only	6.14	6.75	6.63
Virus and NEST ITM	< 3.0	< 3.0	< 3.0
NEST ITM only*	$ \leq 3.0 $	$ \leq 3.0 $	$ \leq 3.0 $

NEST ITM shows cytotoxicity on MDCK cells when diluted to 1:1,000.

NEST ITM must be used at a ratio of at least 1:10 at a minimum of 10 seconds exposure time to demonstrate inactivation of Adenovirus, Influenza A, and Parainfluenza 2 virus. Measuring Adenovirus, Influenza A, and Parainfluenza 2 virus inactivation below 1 x 10 TCIDso was not possible because of the cytotoxic affects NEST ITM has on the cell culture based assay.

5.8 Statement of Substantial Equivalence

The Disposable Sampler Inactivated Transport Media has the same intended use as the PrimeStore MTM predicate device, and the same or similar technological characteristics. The differences in technological characteristics do not raise new or different questions of safety and effectiveness. Performance testing has demonstrated the Disposable Sampler Inactivated Transport Media is as safe and effective as the predicate device. Therefore, the Disposable Sampler Inactivated Transport Media is substantially equivalent to the predicate device.

Regulation Number and Section

§ 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.