(171 days)
Not Found
No
The device description and performance studies focus on the chemical properties of the transport media and the physical design of the collection kit. There is no mention of any computational analysis, algorithms, or learning processes that would indicate the use of AI/ML.
No.
The device is intended for the collection, inactivation, stabilization, and transportation of saliva specimens for diagnostic purposes, not for the treatment or diagnosis of disease in a living being.
No
The device is a collection, inactivation, stabilization, and transportation kit for specimens, but it does not perform any diagnostic function itself. It prepares samples for use with legally marketed molecular diagnostic devices.
No
The device description clearly outlines physical components including a tube, transport media, a funnel, and a cap with a puncture tip. It is a collection kit, not a software application.
Based on the provided information, yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use: The intended use explicitly states that the kit is for the "collection, inactivation, stabilization, and transportation, of unprocessed saliva specimens suspected of containing SARS-CoV-2." It also states that the collected and preserved specimens are "suitable for use with legally marketed molecular diagnostic devices." This clearly indicates its role in the diagnostic process, specifically in preparing samples for downstream diagnostic testing.
- Device Description: The description details how the device functions to preserve and inactivate the sample, making it suitable for laboratory analysis.
- Performance Studies: The inclusion of performance studies like analytical sensitivity (Detection Limit), stability, and inactivation studies are typical for IVD devices to demonstrate their suitability for diagnostic purposes.
- Predicate Device: The mention of a predicate device (K202641; DNA/RNA Shield™ Collection Tube) which is also likely an IVD, further supports the classification of this device as an IVD.
While the device itself doesn't perform the final diagnostic test (it's a collection and preservation kit), its function is integral to the in vitro diagnostic process by preparing the sample for analysis by other legally marketed molecular diagnostic devices.
N/A
Intended Use / Indications for Use
The DNA/RNA Shield™ SafeCollection Kit is intended for the collection, inactivation, stabilization, and transportation, of unprocessed saliva specimens suspected of containing SARS-CoV-2. The DNA/RNA Shield™ SafeCollect Saliva Collection Kit is intended to transport and store saliva specimens at ambient temperature (20-25°C) from the collection site to the laboratory. Specimens collected and preserved in a DNA/RNA Shield™ Saliva Collection kit sample collection tube are suitable for use with legally marketed molecular diagnostic devices.
Product codes
QBD
Device Description
The DNA/RNA Shield SafeCollect Saliva Tube consists of a tube pre-filled with DNA/RNA Shield transport media. DNA/RNA Shield is a transport media that ensures stability of SARS-CoV-2 RNA during sample transport/storage at ambient temperatures (20-25 ℃) and is intended to inactivate SARS-CoV-2, effectively lyses cells from collected saliva specimens.
The DNA/RNA Shield SafeCollect Saliva Tube contains a foil seal barrier that sequesters the DNA/RNA Shield transport media inside of the tube, until the cap, with a Safe Puncture tip is used to seal the DNA/RNA Shield SafeCollect Saliva tube. When the foil seal barrier is broken by the Safe Puncture Tip, the specimen is then, and only then mixed with the DNA/RNA Shield™ transport media.
The DNA/RNA Shield SafeCollect Saliva Collection Kit consists of a DNA/RNA Shield SafeCollect Saliva Tube, a funnel designed for the collection of human saliva samples, and a cap with a Safe Puncture tip. Sample collection is conducted under the supervision of a healthcare provider. The user deposits their saliva into the collection tube with the aid of the attached funnel, the user removes the funnel and replaces it with the cap. Upon twisting and closing the Safe Puncture tip cap, the DNA/RNA Shield is released into the tube and mixes with the saliva.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Saliva (unprocessed saliva specimens)
Indicated Patient Age Range
Not Found
Intended User / Care Setting
Sample collection is conducted under the supervision of a healthcare provider.
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
LoD Determination (Preliminary):
- Sample size: 5 replicates for each concentration.
- Data source: SARS-CoV-2 negative saliva used as a clinical matrix, collected in DNA/RNA Shield SafeCollect Saliva Collection Kit and spiked with inactivated SARS-CoV-2.
- Annotation protocol: Tested with the authorized Quick SARS-CoV-2 rRT-PCR Kit. Preliminary LoD determined as the lowest concentration for which 5/5 independent replicates tested positive.
LoD Determination (Confirmatory):
- Sample size: 20 replicates for each concentration.
- Data source: Inactivated SARS-CoV-2 spiked into negative saliva specimens.
- Annotation protocol: Tested with the authorized Quick SARS-CoV-2 rRT-PCR Kit. Confirmatory LoD was determined as the lowest concentration at which ≥ 19/20 replicates tested positive.
Stability of SARS-CoV-2 in saliva specimens:
- Sample size: 3 replicates for each time point.
- Data source: Negative saliva specimens spiked with 3X LoD of SARS-CoV-2 (750 GEC/ml), collected using the DNA/RNA Shield SafeCollect Saliva Collection Kit and stored at room temperature.
- Annotation protocol: Measured using the Quick SARS-CoV-2 rRT-PCR Kit at day 0, 1, 2, 3, 4, 5, 6, 7, 14, and 21.
Inactivation Study:
- Sample size: Replicated three independent times.
- Data source: SARS-CoV-2 stock titer of 9x10 PFU/mL. Mixed virus with DNA/RNA Shield SafeCollect medium.
- Annotation protocol: Incubated mixture for 30 minutes at room temperature. Serial diluted mixture in cell culture medium and added to confluent monolayers of VeroE6 cells. Cells and media incubated at 37°C in 5% CO2 for one hour. After incubation, overlay media was added, and cells were incubated for two days, then fixed and stained with 0.5% crystal violet for plaque visualization and enumeration. Positive control (untreated viral stock), negative control (media only), and no-virus media control (DNA/RNA Shield media only) were included.
Summary of Performance Studies
Study Type: Analytical sensitivity (Limit of Detection), Stability, Inactivation.
Limit of Detection (LoD) Study:
- Sample Size: Preliminary LoD study used 5 replicates per concentration. Confirmatory LoD study used 20 replicates per concentration.
- Key Results:
- The preliminary LoD was determined to be 83 GEC/mL (5 GEC/rxn), where all 5/5 replicates were positive.
- The final, confirmatory LoD for saliva was determined to be 250 GEC/mL (15 GEC/rxn), as ≥ 19/20 replicates tested positive at this concentration. This is equivalent to the established LoD of the authorized reference assay.
Stability of SARS-CoV-2 in saliva specimens:
- Sample Size: 3 replicates for each time point (Day 0, 1, 2, 3, 4, 5, 6, 7, 14, 21).
- Key Results:
- SARS-CoV-2 spiked at 3X LoD (750 GEC/mL) into negative saliva specimens collected in the kit showed consistent call rates of 3/3 across all tested days up to day 21.
- Average Ct values remained stable, with standard deviations indicating minimal variation.
- There was no significant change in stability over time, and the data were within the acceptance criteria (≤+/-10% deviation from day 0).
- Conclusion: SARS-CoV-2 is stable in saliva specimens collected using the DNA/RNA Shield SafeCollect Saliva Collection kit for up to 21 days when stored at room temperature (20-25 °C).
Inactivation Study:
- Sample Size: Replicated three independent times.
- Key Results:
- The DNA/RNA Shield media showed no cytotoxicity on VeroE6 cells when diluted 1:1,000.
- The mixture of SARS-CoV-2 and DNA/RNA Shield media combined for 30 minutes demonstrated at least a 2-log reduction in SARS-CoV-2.
- Greater than a 2-log reduction could not be quantified due to the starting virus concentration and the necessity to dilute the media 1:1000 to avoid cytotoxicity.
- Conclusion: Zymo DNA/RNA shield inactivates SARS-CoV2 when incubated for at least 30 minutes at room temperature.
Standalone performance, AUC, MRMC: Not Found
Key Metrics
Limit of Detection (LoD): 250 GEC/mL (15 GEC/rxn) for SARS-CoV-2 in saliva specimens.
Stability: No significant change in stability over 21 days at room temperature (20-25 °C), within +/-10% deviation from day 0.
Inactivation Rate: At least a 2-log reduction in SARS-CoV-2 after 30 minutes incubation.
Predicate Device(s)
Reference Device(s)
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information
Not Found
§ 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
September 28, 2023
Image /page/0/Picture/1 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: the Department of Health & Human Services logo on the left and the FDA acronym with the full name of the agency on the right. The FDA part of the logo is in blue, with the acronym in a square and the full name, "U.S. Food & Drug Administration," written out to the right of it.
Zymo Research Julie Ogi Regulatory Office 17062 Murphy Ave Irvine, California 92614
Re: K231013
Trade/Device Name: DNA/RNA Shield SafeCollect Saliva Collection Kit Regulation Number: 21 CFR 866.2950 Regulation Name: Microbial nucleic acid storage and stabilization device Regulatory Class: Class II Product Code: QBD Dated: August 15, 2023 Received: August 29, 2023
Dear Julie Ogi:
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 (the 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 available 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.
Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).
1
Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30, Design controls; 21 CFR 820.90, Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review, the QS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).
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 Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-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 Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 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.
Sincerely,
Noel J. Gerald -S
Noel J. Gerald, Ph.D. Branch Chief Bacterial Respiratory and Medical Countermeasures Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health
2
Indications for Use
510(k) Number (if known) K231013
Device Name
DNA/RNA Shield™ SafeCollect Saliva Collection Kit
Indications for Use (Describe)
The DNA/RNA Shield™ SafeCollection Kit is intended for the collection, inactivation, stabilization, and transportation, of unprocessed saliva specimens suspected of containing SARS-CoV-2. The DNA/RNA Shield™ SafeCollect Saliva Collection Kit is intended to transport and store saliva specimens at ambient temperature (20-25°C) from the collection site to the laboratory. Specimens collected and preserved in a DNA/RNA Shield™ Saliva Collection kit sample collection tube are suitable for use with legally marketed molecular diagnostic devices.
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) |
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3
| Applicant | Zymo Research
17062 Murphy Ave.
Irvine, CA 92614
USA |
|---------------------------------------|----------------------------------------------------------------------------------------------------------------|
| Establishment Registration No. | 3007020052 |
| Contact Person | Julie Ogi
Regulatory Officer
Phone (949) 679-1190
Fax (949) 266-9452
e-mail jogi@zymoresearch.com) |
| Summary Date | March 31, 2023 |
| Proprietary Name | DNA/RNA Shield™ SafeCollect Saliva Collection Kit |
| US Product Codes / Regulation Numbers | QBD - Transport device for the stabilization of microbial
nucleic acids / 21 CRF 866.2950 |
| Classification | Class II |
| Predicate Device | DNA/RNA Shield™ Collection Tube
(K202641) |
Intended Use
The DNA/RNA Shield SafeCollect Saliva Collection Kit is intended for the collection, inactivation, stabilization, and transportation, of unprocessed saliva specimens suspected of containing SARS-CoV-2. The DNA/RNA Shield SafeCollect Saliva Collection Kit is intended to transport and store saliva specimens at ambient temperature (20-25°C) from the collection site to the laboratory. Specimens collected and preserved in a DNA/RNA Shield SafeCollect Saliva Collection kit sample collection tube are suitable for use with legally marketed molecular diagnostic devices.
4
Device Description
The DNA/RNA Shield SafeCollect Saliva Tube consists of a tube pre-filled with DNA/RNA Shield transport media. DNA/RNA Shield is a transport media that ensures stability of SARS-CoV-2 RNA during sample transport/storage at ambient temperatures (20-25 ℃) and is intended to inactivate SARS-CoV-2, effectively lyses cells from collected saliva specimens.
The DNA/RNA Shield SafeCollect Saliva Tube contains a foil seal barrier that sequesters the DNA/RNA Shield transport media inside of the tube, until the cap, with a Safe Puncture tip is used to seal the DNA/RNA Shield SafeCollect Saliva tube. When the foil seal barrier is broken by the Safe Puncture Tip, the specimen is then, and only then mixed with the DNA/RNA Shield™ transport media.
The DNA/RNA Shield SafeCollect Saliva Collection Kit consists of a DNA/RNA Shield SafeCollect Saliva Tube, a funnel designed for the collection of human saliva samples, and a cap with a Safe Puncture tip. Sample collection is conducted under the supervision of a healthcare provider. The user deposits their saliva into the collection tube with the aid of the attached funnel, the user removes the funnel and replaces it with the cap. Upon twisting and closing the Safe Puncture tip cap, the DNA/RNA Shield is released into the tube and mixes with the saliva.
5
Image /page/5/Picture/0 description: The image shows the logo for Zymo Research. The logo consists of three green oval shapes, each containing two smaller circles inside. Below the shapes, the text "ZYMO RESEARCH" is written in bold, followed by the tagline "The Beauty of Science is to Make Things Simple".
Comparison to Predicate Device
| Device & Predicate Device(s): | Device: K231013 | Predicate K202641 |
|---|---|---|
| Device Trade Name | DNA/RNA Shield | |
| SafeCollect Saliva | ||
| Collection Kit | DNA/RNA Shield | |
| Collection Tube | ||
| General Device | ||
| Characteristic Similarities | ||
| Intended Use/Indications For | ||
| Use | The DNA/RNA Shield | |
| SafeCollect Saliva | ||
| Collection Kit is intended | ||
| for the collection, | ||
| inactivation, stabilization, | ||
| and transportation, of | ||
| unprocessed saliva | ||
| specimens suspected of | ||
| containing SARS-CoV-2. | ||
| The DNA/RNA Shield | ||
| SafeCollect Saliva | ||
| Collection Kit is intended | ||
| to transport and store | ||
| saliva specimens at | ||
| ambient temperature (20- | ||
| 25°C) from the collection | ||
| site to the laboratory. | ||
| Specimens collected and | ||
| preserved in a DNA/RNA | ||
| Shield™ SafeCollect | ||
| Saliva Collection kit | ||
| sample collection tube are | ||
| suitable for use with | ||
| legally marketed | ||
| molecular diagnostic | ||
| devices. | The DNA/RNA Shield | |
| collection tube is intended | ||
| for the stabilization and | ||
| inactivation of upper and | ||
| lower respiratory human | ||
| specimens suspected of | ||
| containing SARS-CoV-2. | ||
| These devices can be used | ||
| for collection transport | ||
| and storage of specimens | ||
| at ambient temperatures | ||
| (20-25°C). Specimens | ||
| collected and stored in a | ||
| DNA/RNA Shield | ||
| collection tube are | ||
| suitable for use with | ||
| legally marketed | ||
| molecular diagnostic | ||
| devices. | ||
| Analyte | SARS-CoV-2 RNA | Same |
| Sample stability | 20-25°C | Same |
| Collection media | DNA/RNA Shield media | Same |
| Special conditions for use | For prescription use only | |
| For in-vitro diagnostic use | ||
| only | Same | |
| General Device | ||
| Characteristic Differences | ||
| Sample collection | Saliva | Upper and lower |
| respiratory specimens | ||
| RNA Stabilization at room | ||
| temperature | Saliva: up to 21 days | Upper/lower respiratory |
| tract samples: up to 28 | ||
| days |
6
Image /page/6/Picture/0 description: The image shows the logo for Zymo Research. The logo consists of three green oval shapes arranged in a triangular pattern, with the words "ZYMO RESEARCH" printed in bold black letters below. Underneath the company name is the tagline "The Beauty of Science is to Make Things Simple" in a smaller, italicized font.
Performance data
- a) Detection Limit:
An analytical sensitivity study was conducted to determine the Limit of Detection (LoD) of SARS-CoV-2 when detected in saliva samples collected using the DNA/RNA Shield SafeCollect Saliva Collection kit in combination with the authorized Quick SARS-CoV-2 rRT-PCR Kit for SARS-CoV-2 detection. To determine a preliminary LoD, SARS-CoV-2 negative saliva was used as a clinical matrix collected in DNA/RNA Shield SafeCollect Saliva Collection Kit and spiked with inactivated SARS-CoV-2. The preliminary LoD was determined as the lowest concentration for which 5/5 independent replicates tested positive. Results for the preliminary LoD determination are shown in Table 1 below.
Table 1: Preliminary LoD Determination
| Concentrations Tested (GEC/mL) | Replicates | SARS-CoV-2 Targets | Interpretation | Call Rate |
|---|---|---|---|---|
| 8.3 x 104 | ||||
| (5,000 GEC/rxn) | 1 | 27.33 | Positive | 5/5 |
| 2 | 28.03 | Positive | ||
| 3 | 27.73 | Positive | ||
| 4 | 27.60 | Positive | ||
| 5 | 27.73 | Positive | ||
| 8.3 x 103 | ||||
| (500 GEC/rxn) | 1 | 29.77 | Positive | 5/5 |
| 2 | 28.83 | Positive | ||
| 3 | 29.33 | Positive | ||
| 4 | 29.64 | Positive | ||
| 5 | 29.97 | Positive | ||
| 8.3 x 102 | ||||
| (50 GEC/rxn) | 1 | 32.37 | Positive | 5/5 |
| 2 | 32.95 | Positive | ||
| 3 | 32.31 | Positive | ||
| 4 | 32.88 | Positive | ||
| 5 | 32.49 | Positive | ||
| 83 | ||||
| (5 GEC/rxn) | 1 | 39.54 | Positive | 5/5 |
| 2 | 34.16 | Positive | ||
| 3 | 38.87 | Positive | ||
| 4 | 32.54 | Positive | ||
| 5 | 33.36 | Positive | ||
| 8.3 | ||||
| (0.5 GEC/rxn) | 1 | N/A | Negative | 1/5 |
| 2 | 44.00 | Inconclusive | ||
| 3 | 37.89 | Positive | ||
| 4 | 40.94 | Inconclusive | ||
| 5 | N/A | Negative | ||
| Concentrations Tested (GEC/mL) | Replicates | SARS-CoV-2 Targets | Interpretation | Call Rate |
| 250 | ||||
| (15 GEC/ rxn) | 1 | 34.62 | Positive | 19/20 |
| 2 | 34.83 | Positive | ||
| 3 | 33.99 | Positive | ||
| 4 | N/A | Negative | ||
| 5 | 35.84 | Positive | ||
| 6 | 33.94 | Positive | ||
| 7 | 35.25 | Positive | ||
| 8 | 35.01 | Positive | ||
| 9 | 33.99 | Positive | ||
| 10 | 33.69 | Positive | ||
| 11 | 34.01 | Positive | ||
| 12 | 34.03 | Positive | ||
| 13 | 35.32 | Positive | ||
| 14 | 34.46 | Positive | ||
| 15 | 35.84 | Positive | ||
| 16 | 35.56 | Positive | ||
| 17 | 33.76 | Positive | ||
| 18 | 35.33 | Positive | ||
| 19 | 34.00 | Positive | ||
| 20 | 33.56 | Positive | ||
| 166 | ||||
| (10 GEC/ rxn) | 1 | 35.57 | Positive | 17/20 |
| 2 | 35.35 | Positive | ||
| 3 | 35.78 | Positive | ||
| 4 | N/A | Negative | ||
| 5 | 33.80 | Positive | ||
| 6 | 34.42 | Positive | ||
| 7 | 35.19 | Positive | ||
| 8 | 35.01 | Positive | ||
| 9 | 34.87 | Positive | ||
| 10 | 36.17 | Positive | ||
| 11 | N/A | Negative | ||
| 12 | 34.25 | Positive | ||
| 13 | 37.80 | Positive | ||
| 14 | N/A | Negative | ||
| 15 | 34.64 | Positive | ||
| 16 | 35.44 | Positive | ||
| 17 | 35.51 | Positive | ||
| 18 | 35.46 | Positive | ||
| 19 | 36.05 | Positive | ||
| 20 | 34.66 | Positive | ||
| 83 | ||||
| (5 GEC/ rxn) | 1 | 41.48 | Inconclusive | 13/20 |
| 2 | 37.02 | Positive | ||
| 3 | 37.57 | Positive | ||
| 4 | 37.07 | Positive | ||
| 5 | N/A | Negative | ||
| 6 | 36.54 | Positive | ||
| 7 | 37.17 | Positive | ||
| 8 | 37.25 | Positive |
To confirm the LoD, inactivated SARS-CoV-2 was spiked into negative saliva specimens and 20 replicates were independently processed. The lowest concentration at which all 5 replicates were positive in the preliminary LoD (i.e., 83 GEC/mL) was used as a starting point for the confirmatory LoD study. Therefore, concentrations above 83 GEC/mL (increasing by factor 2) were tested until ≥ 19/20 replicates tested positive. The final LoD for saliva was determined to be at the lowest concentration at which ≥ 19/20 replicates test positive. The final LoD was determined to be 250 GEC/mL (15 GEC/rxn). Results of the confirmatory LoD study for saliva specimens can be seen in Table 2 below.
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Image /page/7/Picture/0 description: The image shows the logo for Zymo Research. The logo consists of three green oval shapes arranged in a triangular pattern, with the words "ZYMO RESEARCH" printed in bold black letters below. Underneath the company name is the tagline "The Beauty of Science is to Make Things Simple".
8
Image /page/8/Picture/0 description: The image shows the logo for Zymo Research. The logo consists of three green oval shapes arranged in a triangular pattern. Below the shapes, the text "ZYMO RESEARCH" is written in a bold, sans-serif font. Underneath the company name is the tagline "The Beauty of Science is to Make Things Simple".
| 9 | 35.21 | Positive |
|---|---|---|
| 10 | N/A | Negative |
| 11 | 37.08 | Positive |
| 12 | N/A | Negative |
| 13 | 35.57 | Positive |
| 14 | N/A | Negative |
| 15 | N/A | Negative |
| 16 | N/A | Negative |
| 17 | 34.74 | Positive |
| 18 | 35.01 | Positive |
| 19 | 35.03 | Positive |
| 20 | 35.54 | Positive |
Conclusion:
The DNA/RNA Shield SafeCollect medium used to collect saliva, and the Authorized Quick SARS-CoV-2 2rRT-PCR Kit reached a SARS-CoV-2 LoD of 250 GEC/mL (15 GEC/reaction) for saliva specimens, which is equivalent to the established LoD of the authorized reference assay.
b) Stability of SARS-CoV-2 in saliva specimens:
The room temperature (20-25 °C) stability of SARS-CoV-2 in DNA/RNA Shield SafeCollect Saliva kit was established by spiking 3X LoD of SARS-CoV-2 (750 GEC/ml) (see LoD section above) into negative saliva specimens collected using the DNA/RNA Shield SafeCollect Saliva Collection Kit and stored at room temperature in a time course study of 21 days. The room temperature stability of SARS-CoV-2 was measured using the Quick SARS-CoV-2 rRT-PCR Kit at day 0, 1, 2, 3, 4, 5, 6, 7, 14, and 21. Results are summarized in Table 3
| Concentration Tested | Days at Room Temperature | Replicates | Average Ct (Standard Deviation) | Call Rate |
|---|---|---|---|---|
| Low Positive | ||||
| 3x LoD | ||||
| (750 GEC/mL) | Day 0 | 3 | 33.75 (0.21) | 3/3 |
| Day 1 | 3 | 33.37 (0.22) | 3/3 | |
| Day 2 | 3 | 34.10 (0.35) | 3/3 | |
| Day 3 | 3 | 33.93 (0.32) | 3/3 | |
| Day 4 | 3 | 34.24 (0.97) | 3/3 | |
| Day 5 | 3 | 34.34 (0.24) | 3/3 | |
| Day 6 | 3 | 34.75 (0.87) | 3/3 | |
| Day 7 | 3 | 34.44 (0.41) | 3/3 | |
| Day 14 | 3 | 33.98 (0.36) | 3/3 | |
| Day 21 | 3 | 34.05 (0.48) | 3/3 |
Table 3. SARS-CoV-2 Stability
Conclusion:
There is no significant change in stability over time and the data are within the acceptance criteria (≤+/-10% deviation from day 0). Stability is determined by acceptable data that supports the stability claim and does not exceed day 0 by a 3-log range. Therefore, SARS-CoV-2 is stable in saliva specimens collected using the DNA/RNA Shield SafeCollect Saliva Collection kit for up to 21 days when stored at room temperature (20-25 °C).
c) Inactivation:
An inactivation study was conducted to test DNA/RNA Shield SafeCollect Saliva Collection kit media ability to inactivate SARS-CoV-2. The inactivation study used a stock titer of 9x10 PFU/mL for SARS-CoV-2. The study was performed in a Biosafety Level 3 (BSL-3) facility.
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mixed with 300 uL of DNA/RNA Shield SafeCollect medium). The virus and media mixture were incubated for 30 mins at room temperature. The mixture was then serial diluted in cell culture medium and added to confluent monolavers of VeroE6 cells. Cells and media were incubated at 37°C in 5% CO2 for one hour with gentle rocking every 15 minutes. Media was aspired and 1.0 mL of pre-warmed overlay media was added. Cells were incubated for two days, fixed in 10% formaldehyde, and stained with 0.5% crystal violet to enable plaque visualization and enumeration. A positive control (untreated viral stock) and negative control (media only) were included in each run. A no-virus media control (DNA/RNA Shield media only) was also included to assess whether the media was cytotoxic. The study was replicated three independent times.
Inactivation rate:
The DNA/RNA Shield media showed no cytotoxicity on veroE6 cells when the media was diluted 1:1,000 with cell culture media. The 1:1.000 dilution factor is needed to avoid cytotoxic effects the DNA/RNA Shield media has on the cell monolayer. The mixture of SARS-CoV-2 and DNA/RNA Shield media combined for 30 minutes demonstrated at least a 2-log reduction in SARS-CoV-2. Greater than a 2-log reduction of SARS-CoV-2 could not be quantified based on the starting concentration of the virus compounded by the need to dilute the DNA/RNA Shield media 1:1000. CPE could not be observed at