Spectrum Saliva Collection Device

K223497 · Spectrum Solutions, LLC · QBD · Feb 17, 2023 · Microbiology

Device Facts

• Record ID: K223497
• Device Name: Spectrum Saliva Collection Device
• Applicant: Spectrum Solutions, LLC
• Product Code: QBD · Microbiology
• Decision Date: Feb 17, 2023
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.2950
• Device Class: Class 2

Indications for Use

The Spectrum Solutions Saliva Collection Device is designed for use in the non-invasive collection, inactivation, and stabilization of viral nucleic acids for in vitro diagnostic testing of saliva samples. The device is intended to inactivate and stabilize human clinical saliva samples from the collection site to the laboratory at room temperature. The saliva sample is stabilized and suitable for use with legally marketed molecular diagnostic devices. The device is intended to be used by a health care provider for samples suspected of containing SARS-CoV-2.

Device Story

Device consists of plastic tube, funnel, and cap with stem flare containing inactivating media. Used by healthcare providers to collect human saliva samples. User deposits saliva into tube via funnel; removing funnel and closing cap releases stabilizing solution into sample. Media lyses lipid membranes, inactivates enzymes, and stabilizes viral nucleic acids. Samples stored at room temperature for transport to laboratory. Output is stabilized saliva sample suitable for molecular diagnostic testing (e.g., RT-PCR). Benefits include safe handling of potentially infectious samples due to viral inactivation and preservation of nucleic acid integrity for diagnostic accuracy.

Clinical Evidence

No clinical data. Bench testing only. Stability study (n=4) confirmed SARS-CoV-2 RNA integrity over 28 days at 20-25°C using TaqPath COVID-19 assay. LoD established at 250 GEC/ml (20 GEC/reaction) with >95% accuracy. Inactivation study demonstrated >4 log10 reduction of SARS-CoV-2 titer within 10 seconds.

Technological Characteristics

Medical-grade polypropylene tube, funnel, and cap. Inactivating media contains guanidine thiocyanate, ethanol, EDTA, and blue dye. Non-sterile, single-use. No electronic components or software.

Indications for Use

Indicated for the non-invasive collection, inactivation, and stabilization of viral nucleic acids from human saliva samples suspected of containing SARS-CoV-2, for use by healthcare providers in conjunction with legally marketed molecular diagnostic devices.

Regulatory Classification

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.

Special Controls

The special controls for this device are: (1) The intended use for the 21 CFR 809.10 labeling must include a detailed description of microorganisms and types of human specimens intended to be preserved. (2) The 21 CFR 809.10(b) labeling must include: (i) A detailed device description, including all device components. (ii) Performance characteristics from applicable analytical studies, including but not limited to, 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. (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, but not limited to, extraction platforms and assays used for the detection of preserved nucleic acids. (iii) The limit of detection (LoD) of the molecular test used to establish microorganism nucleic acid stability.

*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.

Predicate Devices

• Zymo Research DNA/RNA Shield Collection Tube (K202641)

Submission Summary (Full Text)

{0}

FDA

U.S. FOOD & DRUG

ADMINISTRATION

# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

INSTRUMENT ONLY

## I Background Information:

A 510(k) Number

K223497

B Applicant

Spectrum Solutions, LLC

C Proprietary and Established Names

Spectrum Saliva Collection Device

D Regulatory Information

| Product Code(s) | Classification | Regulation Section | Panel |
| --- | --- | --- | --- |
| QBD | Class II | 21 CFR 866.2950 - Microbial Nucleic Acid Storage And Stabilization Device | MI - Microbiology |

## II Submission/Device Overview:

A Purpose for Submission:

To make a substantial equivalence determination for the Spectrum Solutions Saliva Collection Device for the collection, transport, and storage of viral specimens in saliva to the laboratory for downstream testing.

B Measurand:

Storage and stability of nucleic acids from SARS-CoV-2

C Type of Test:

Microbial nucleic acid storage and stabilization device

## III Intended Use/Indications for Use:

Food and Drug Administration

10903 New Hampshire Avenue

Silver Spring, MD 20993-0002

www.fda.gov

{1}

# A Intended Use(s):

See Indications for Use below.

# B Indication(s) for Use:

The Spectrum Solutions Saliva Collection Device is designed for use in the non-invasive collection, inactivation, and stabilization of viral nucleic acids for in vitro diagnostic testing of saliva samples. The device is intended to inactivate and stabilize human clinical saliva samples from the collection site to the laboratory at room temperature. The saliva sample is stabilized and suitable for use with legally marketed molecular diagnostic devices. The device is intended to be used by a health care provider for samples suspected of containing SARS-CoV-2.

# C Special Conditions for Use Statement(s):

For Prescription Use Only

# D Special Instrument Requirements:

None

# IV Device/System Characteristics:

# A Device Description:

The Saliva Collection Device consists of a plastic tube designed for the collection of human saliva samples, a funnel, a cap with a stem flare, and a fluid chamber containing Spectrum's inactivating media. Sample collection is conducted under the supervision of a Health care 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 cap, the inactivating media is released into the tube and mixes with the saliva.

Figure 1. Saliva Collection Device Sample Collection Steps

# B Principle of Operation:

The components of the inactivating media for the Spectrum device are intended to inactivate SARS CoV-2 capsids, disrupt/lyse membranes, denature proteins, inactivate enzymes, and stabilize SARS CoV-2 RNA. The inactivating solution includes buffering salts, purified water, and surfactants as well as the following:

K223497 - Page 2 of 10

{2}

- Guanidine Thiocyanate
- Ethanol
- EDTA
- Blue dye

## C Instrument Description Information:

1. Instrument Name:
N/A

2. Specimen Identification:
N/A

3. Specimen Sampling and Handling:
N/A

4. Calibration:
N/A

5. Quality Control:
N/A

## V Substantial Equivalence Information:

### A Predicate Device Name(s):
Zymo Research DNA/RNA Shield Collection Tube

### B Predicate 510(k) Number(s):
K202641

## C Comparison with Predicate(s):

| Feature: | Device: K223497 | Predicate: K202641 |
| --- | --- | --- |
| Device Trade Name | Spectrum Solutions Saliva Collection Device | DNA/RNA Shield Collection Tube |
| General Device Characteristic Similarities | | |
| Intended Use/Indications For Use | The Spectrum Solutions Saliva Collection Device is designed for use in the non-invasive collection, | The DNA/RNA Shield collection tube is intended for the stabilization and inactivation of upper and |

K223497 - Page 3 of 10

{3}

K223497 - Page 4 of 10
| | inactivation, and stabilization of viral nucleic acids for in vitro diagnostic testing of saliva samples. The device is intended to inactivate and stabilize human clinical saliva samples from the collection site to the laboratory at room temperature. The saliva sample is stabilized and suitable for use with legally marketed molecular diagnostic devices. The device is intended to be used by a health care provider for samples suspected of containing SARS-CoV-2. | 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. |
| --- | --- | --- |
| Additive/Reagent | Nucleic Acid Stabilization solution | Same |
| Analyte | RNA from SARS-CoV-2 | Same |
| Limit of Detection | 250 GEC/ml (20 GEC/reaction) | Same |
| Specimen Stability | SARS-CoV-2 for 28 days at 20-25 °C | Same |
| Transport Media | Disrupt/lyses lipid membranes, inactivates enzymes, and stabilizes nucleic acids | Same |
| Material | Medical-grade polypropylene | Same |
| Sterility | Non-Sterile | Same |
| General Device Characteristic Differences | | |
| Specimen Type | Saliva only | Upper, lower respiratory, and saliva |
| Device Material | Tube - Polypropylene Tube and Funnel containing Spectrum’s Nucleic Acid Stabilization Solution and a Polypropylene Cap. | Plastic Collection Tube pre-filled with DNA/RNA Shield transport media. |
| Inactivation test | Greater than 4 Log viral reduction in 10 seconds | Greater than 2 log viral reduction in 30 minutes |

{4}

VI Standards/Guidance Documents Referenced:

Special controls that are applicable to regulation 21 CFR 866.2950

VII Performance Characteristics (if/when applicable):

A Analytical Performance:

1. Precision/Reproducibility:
N/A

2. Linearity:
N/A

3. Analytical Specificity/Interference:
N/A

4. Assay Reportable Range:
N/A

5. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods):

Shelf life: The shelf life for the Spectrum Saliva Collection Device is 24 months at room temperature (15-30°C) after the date of manufacture when stored at 5-30°C. The stability of the Spectrum Saliva Collection Device was performed using real-time analysis on three (3) lots of device caps (tube tops) containing inactivating media. All of the 24-month-aged caps tested with inactivating media met the acceptance criteria for stability with regards to color appearance of dye (blue), pH, assays for guanidine thiocyanate, denatured alcohol, and EDTA, specific gravity, bacterial and fungal contamination, and weight. Analytical testing was run on finished commercial products after two years.

Sterilization: The Spectrum Solutions Saliva Collection Devices 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.

Sample Stability: Studies were conducted to evaluate the stability of SARS-CoV-2 in the Spectrum inactivating media for up to 28 days. Contrived samples containing SARS-CoV-2 at a concentration of 3x LoD (62.5 GEC/reaction; 1562.5 GEC/ml) were prepared in saliva matrix collected from a single negative donor. Sample stability was evaluated in a study that utilized two different device lots to collect two samples each (for a total of 4 test samples) that were then stored at ambient temperature (20-25°C) for increasing lengths of time. Aliquots of each contrived sample was tested for SARS-CoV-2 using the TaqPath COVID-19 assay at T=0 and on Days 1, 2, 3, 4, 5, 6, 7, 14, 21, and 28. Aliquots of samples were removed and stored at -20°C at various times and then all aliquots (total of 44 samples) were

K223497 - Page 5 of 10

{5}

tested as a single testing batch. The acceptance criteria for the stability study included a deviation of no more than $+ / - 3$ Ct for each target at given time point from $\mathrm{T} = 0$

Results of stability testing are shown below in Tables 1 and 2.

Table 1. Specimen Stability Study Results

| Time | Orf1ab Ct average | Orf1ab Ct SD | Orf1ab Positive Count | N gene Ct average | N gene Ct SD | N gene Positive Count | S gene Ct average | S gene Ct SD | S gene Positive Count |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| T0 | 33.1 | 0.98 | 4/4 | 31.7 | 0.32 | 4/4 | 32.8 | 0.34 | 4/4 |
| D1 | 31.9 | 0.85 | 4/4 | 31.3 | 0.45 | 4/4 | 33.0 | 0.49 | 4/4 |
| D2 | 32.4 | 0.33 | 4/4 | 31.4 | 0.33 | 4/4 | 33.8 | 0.61 | 4/4 |
| D3 | 32.4 | 0.40 | 4/4 | 31.4 | 0.35 | 4/4 | 33.6 | 0.26 | 4/4 |
| D4 | 32.7 | 1.23 | 4/4 | 31.6 | 0.46 | 4/4 | 34.0 | 1.74 | 4/4 |
| D5 | 32.7 | 0.47 | 4/4 | 31.5 | 0.28 | 4/4 | 34.2 | 0.86 | 4/4 |
| D6 | 32.7 | 0.43 | 4/4 | 31.6 | 0.29 | 4/4 | 33.4 | 0.47 | 3/4 |
| D7 | 33.2 | 0.87 | 4/4 | 31.8 | 0.32 | 4/4 | 34.5 | 1.15 | 4/4 |
| D14 | 33.7 | 0.84 | 4/4 | 32.1 | 0.19 | 4/4 | 33.8 | NA | 1/4 |
| D21 | 34.4 | 0.79 | 4/4 | 33.1 | 0.68 | 4/4 | NA | NA | 0/4 |
| D28 | 33.4 | 3.25 | 4/4 | 34.4 | 0.53 | 4/4 | NA | NA | 0/4 |

Table 2. Specimen Stability Study - Ct Changes Observed

| Time | Orf1ab Ct change from T0 | N gene Ct change from T0 | S gene Ct change from T0 |
| --- | --- | --- | --- |
| T0 | NA | NA | NA |
| D1 | -1.25 | -0.38 | 0.29 |
| D2 | -0.73 | -0.34 | 1.04 |
| D3 | -0.72 | -0.29 | 0.85 |
| D4 | -0.47 | -0.15 | 1.19 |
| D5 | -0.43 | -0.26 | 1.45 |
| D6 | -0.47 | -0.10 | 0.69 |
| D7 | 0.05 | 0.05 | 1.75 |
| D14 | 0.52 | 0.38 | 1.04 |
| D21 | 1.28 | 1.40 | ND |
| D28 | 0.24 | 2.71 | ND |

43/44 of the samples tested positive for SARS-CoV-2 at all time points during initial testing. One Day 21 sampled failed to produce a conclusive result when first tested, but upon retesting the sample (per the TaqPath COVID-19 protocol) the sample gave a positive result. All samples gave expected internal control results (amplification of MS2 phage nucleic acid) to monitor the integrity of nucleic acid extractions and RT-PCR for each sample. There were minimal Ct changes over the 28 days of the experiment with the highest deviation shown for N gene amplification on day 28, however the observed difference was still within the study's acceptance criteria. The S gene target did show a decreased overall reactivity, however this is consistent and expected due to SARS-CoV-2 viral mutation as discussed above. Since the

K223497 - Page 6 of 10

{6}

TaqPath COVID-19 assay requires 2/3 targets to amplify for positive results, this decreased reactivity did not impact assay performance.

These test results fall within the established acceptance criteria and indicate samples are stable in the Spectrum inactivating media for up to 28 days.

# 6. Detection Limit:

Limit of Detection (LoD) testing was conducted to determine the lowest concentration of whole genome (Genome Equivalent Copies; GEC) that contains measurable nucleic acids that can be repeatedly recovered from the transport media with greater than $95\%$ accuracy. LoD was obtained by spiking the Spectrum SDNA-1000 collecting device containing negative matrix (saliva + preservative) with various concentrations of SARS-CoV-2 before performing RNA extractions using the MagMAX Viral/Pathogen II (MVP II) Nucleic Acid Isolation Kit on the KingFisher Flex Purification System. Viral RNA was subsequently amplified using the TaqPath COVID-19 Combo Kit using the Applied Biosystems QuantStudio 5 (QS5) Fast Real-Time PCR System. The RNA extraction and RT-PCR kits contained an internal MS2 Phage control to monitor the integrity of nucleic acid extraction and RT-PCR for each specimen. Ct values and detection of virus were determined using the EUA IFU guidelines and the Applied Biosystems COVID-19 Interpretative Software v2.5.

For the COVID-19 TaqPath assay, Ct values $< 37$ are considered positive for SARS-CoV-2. Ct values of 37 to $< 40$ were considered inconclusive and those samples retested to determine SARS-CoV-2 status, as per the protocol Qualitative Detection of Nucleic Acid from SARS-CoV-2 using the COVID-19 TaqPath Combo Kit (JB-PRO-000105, v.2.1). Three gene targets in SARS-CoV-2 were subjected to amplification (ORF1ab, N gene, and S gene), with a sample considered positive if 2/3 of these gene targets were amplified.

Preliminary LoD testing was initially performed by spiking multiple concentrations of SARS-CoV-2 into clinically negative matrix (saliva) to achieve final concentrations of 0, 10, 20, 50, 100, 250, and 500 GEC/reaction. Five samples of each of these contrived concentrations were tested with results shown below in Table 3.

Table 3. Preliminary LoD Estimation Results

| GEC/reaction | Orf1ab detection | ORF1a Mean Ct | N gene detection | N gene Mean Ct | S gene detection | S gene Mean Ct | SARS-CoV-2 positive result |
| --- | --- | --- | --- | --- | --- | --- | --- |
| 0 | 0/5 | ND | 0/5 | ND | 0/5 | ND | 0/5 |
| 10 | 5/5 | 34.45 | 0/5 | ND | 0/5 | ND | 5/5 |
| 20 | 5/5 | 32.89 | 5/5 | 33.97 | 0/5 | ND | 5/5 |
| 50 | 5/5 | 31.81 | 5/5 | 31.90 | 5/5 | 33.45 | 5/5 |
| 100 | 5/5 | 30.43 | 5/5 | 30.30 | 5/5 | 31.66 | 5/5 |
| 250 | 5/5 | 29.44 | 5/5 | 29.06 | 5/5 | 30.13 | 5/5 |
| 500 | 5/5 | 29.05 | 5/5 | 28.84 | 5/5 | 29.67 | 5/5 |

The estimated LoD based on the preliminary results was identified as 20 GEC/reaction, but one of the three gene targets was successfully amplified (ORF1ab) results from the 10

K223497 - Page 7 of 10

{7}

GEC/reaction dilution. To further confirm this preliminary LoD assessment, both 15 and 20 GEC/reactions were evaluated in a set of 20 replicates (see Table 4). A positive result for a sample was defined as amplification of at least 2 of the 3 gene targets from SARS-CoV-2 (ORF1ab, N gene, S gene) per the IFU from the authorized device.

Confirmatory testing supported an LoD of 20 GEC/reaction with 20/20 replicates for amplification of ORF1ab and 20/20 replicates for N gene, which met the pre-defined criteria for RT-PCR reactions (greater than $95\%$ accuracy for detecting 2/3 targets in samples). However, for the 15 GEC/reaction dilution only 16/20 samples met the requirement of 2/3 targets amplified ( $80\%$ accuracy), which fell below the pre-defined criteria. Therefore, 20 GEC/reaction is the established LoD.

Table 4. Confirmatory LoD Study Results

| GEC/reaction | Orf1ab detection | ORF1a Mean Ct | N gene detection | N gene Mean Ct | S gene detection | S gene Mean Ct | SARS-CoV-2 positive result |
| --- | --- | --- | --- | --- | --- | --- | --- |
| 15 | 20/20 | 33.92 | 16/20 | 35.29 | 0/20 | ND | 0/5 |
| 20 | 20/20 | 33.81 | 20/20 | 35.23 | 2/20 | 36.77 | 5/5 |

Ct values for each of the three amplified genes of the 20 replicates of the confirmatory tests for the 20 GEC/reaction concentration are shown in Table 5.

Table 5. LoD Confirmation Study - Individual Replicate Ct Values

K223497 - Page 8 of 10

{8}

| | Ct Value | | |
| --- | --- | --- | --- |
| Replicate | ORF1ab | N gene | S gene |
| 1 | 36.14 | 34.83 | |
| 2 | 33.09 | 35.84 | |
| 3 | 33.35 | 36.09 | |
| 4 | 30.03 | 36.79 | |
| 5 | 34.50 | 34.48 | |
| 6 | 32.18 | 36.48 | |
| 7 | 34.60 | 34.79 | 36.66 |
| 8 | 33.55 | 34.42 | |
| 9 | 33.86 | 35.72 | |
| 10 | 34.71 | 36.63 | |
| 11 | 34.06 | 33.37 | |
| 12 | 34.24 | 36.01 | 36.87 |
| 13 | 33.85 | 34.47 | 37.61 |
| 14 | 32.80 | 35.45 | 37.83 |
| 15 | 34.17 | 34.02 | |
| 16 | 33.73 | 34.94 | 37.64 |
| 17 | 33.93 | 36.54 | 39.65 |
| 18 | 35.94 | 34.37 | |
| 19 | 33.15 | 35.12 | |
| 20 | 34.30 | 34.28 | |
| AVG: | 33.81 | 35.23 | 37.71 |
| SD: | 1.29 | 0.99 | 1.06 |

Note: The 69-70del mutation interferes with the detection of the S gene target by the TaqPath COVID-19 assay, which may have rendered the S gene target as not detected ("S gene target failure"). But due to the multi-target test design, the overall test performance of the TaqPath assay is not impacted.

In summary, the testing performed established 250 GEC/ml (20 GEC/reaction) as the LoD or the lowest concentration tested that resulted in $>95\%$ accuracy of detection of contrived samples.

# 7. Inactivation

An inactivation study was conducted to verify that the Spectrum SDNA-1000 collecting device inactivates SARS-CoV-2 as efficiently as the predicate device. Specifically, viral inactivation testing of SARS-CoV-2 was conducted using a Virucidal Suspension Test. Viral preparations containing SARS-CoV-2 isolate (B.1.1.529 Omicron variant) of $3.2 \times 10^{6}$ $\mathrm{TCID}_{50}$ (approximately $2.24 \times 10^{6}$ PFU) or lower were combined with various combinations of Spectrum inactivating media and/or neutralization fluid (for the inactivating media).

Following treatment of virus, 10-fold dilutions were made of each preparation and incubated with Vero E6 kidney cells grown to $80 - 90\%$ confluency. After a one hour incubation with the treatments listed above, cells were returned to culture conditions and incubated before cytopathic effects were determined visually with an inverted compound microscope.

K223497 - Page 9 of 10

{9}

The acceptance criteria for the inactivation study included at least a $4\log_{10}$ of $\mathrm{TCID}_{50}$ would be recovered from the virus control (no inactivation) and at least a $3\log_{10}$ reduction in titer could be demonstrated in Spectrum media

Three different lots of devices were tested for viricidal activity, with each test performed in two replicates per device. Plating of each replicate was performed four times.

Under the experimental conditions described above the Spectrum inactivating media inactivates SARS-CoV-2 by an average of $\geq 4.00\log_{10}(>99.99\%)$ following a 10-second exposure and by an average of $\geq 4.00\log_{10}(>99.99\%)$ following a 60-minute exposure.

8. Assay Cut-Off:
N/A

9. Accuracy (Instrument):
N/A

10. Carry-Over:
N/A

B Comparison Studies:
N/A

C Clinical Studies:
N/A

D Clinical Cut-Off:
N/A

E Expected Values/Reference Range:
N/A

F Other Supportive Instrument Performance Characteristics Data:
N/A

VIII Proposed Labeling:
The labeling supports the finding of substantial equivalence for this device.

IX Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

K223497 - Page 10 of 10