The Accula™ Strep A Test performed on the Accula Dock is a molecular in vitro diagnostic test utilizing polymerase chain reaction (PCR) and lateral flow technologies for the qualitative, visual detection of Streptococcus pyogenes (Group A B-hemolytic Streptococcus, Strep A) bacterial nucleic acid. It is intended to aid in the rapid diagnosis of Group A Streptococcus bacterial infections from throat swabs of patients with signs and symptoms of pharyngitis.

All negative test results should be confirmed by bacterial culture because negative results do not preclude infection with Group A Streptococcus and should not be used as the sole basis for treatment.

Device Description

The Accula™ Strep A Test is a semi-automated, colorimetric polymerase chain reaction (PCR) nucleic acid amplification test to qualitatively detect Streptococcus pyogenes (Group A Bhemolytic Streptococcus, Strep A) bacterial nucleic acid from unprocessed throat swabs that have not undergone prior nucleic acid extraction. The system integrates nucleic acid extraction, a novel Mesa Biotech PCR nucleic acid amplification technology named OscAR™, and hybridization-based visual detection into a completely self-contained and automated system. The Accula Strep A system consists of a small reusable Dock to drive the automated testing process, and a single-use disposable test cassette that contains all the enzymes and reagents.

AI/ML Overview

The Mesa Biotech Accula Strep A Test is an in vitro diagnostic test for the qualitative, visual detection of Streptococcus pyogenes (Group A B-hemolytic Streptococcus, Strep A) bacterial nucleic acid from throat swabs. The device integrates nucleic acid extraction, OscAR™ PCR amplification technology, and hybridization-based visual detection.

Acceptance Criteria and Device Performance:

The primary performance metrics for the Accula Strep A Test were Sensitivity, Specificity, Positive Percent Agreement (PPA), and Negative Percent Agreement (NPA). These were evaluated against a reference bacterial culture and an FDA-cleared molecular comparator.

Metric	Acceptance Criteria (Implied)	Reported Device Performance (vs. Blood Agar Culture)	Reported Device Performance (vs. Molecular Comparator)
Sensitivity	High, expected to be comparable to or better than predicate	96.2% (126/131) (95% CI: 91.4%-98.4%)	N/A (PPA used for molecular comparison)
Specificity	High, expected to be comparable to or better than predicate	97.5% (510/523) (95% CI: 95.8%-98.5%)	N/A (NPA used for molecular comparison)
Positive Percent Agreement	High, expected to be comparable to or better than predicate	N/A	93.8% (137/146) (95% CI: 88.7%-96.7%)
Negative Percent Agreement	High, expected to be comparable to or better than predicate	N/A	99.8% (501/502) (95% CI: 98.9%-100%)
Reproducibility (Low Positive)	High agreement (e.g., >95%) across sites, operators, and days	98.9% (89/90) (95% CI: 94.0%-99.8%)	N/A
Reproducibility (Moderate Positive)	High agreement (e.g., >95%) across sites, operators, and days	97.8% (87/89) (95% CI: 92.2%-99.4%)	N/A
Reproducibility (Negative)	High agreement (e.g., >95%) across sites, operators, and days	97.8% (88/90) (95% CI: 92.3%-99.4%)	N/A
Limit of Detection	Expected to detect Strep A at low concentrations	BAA-946: 75 CFU/mL, ATCC 19615: 10 CFU/mL	N/A
Analytical Reactivity	100% detection of tested Strep A strains at appropriate levels	100% detection for 3/4 strains at 1.5x LoD, 100% for all at 3.0x LoD	N/A
Analytical Specificity	No cross-reactivity with common respiratory pathogens and flora	All 47 tested organisms showed 0/3 positive when Strep A absent; 3/3 positive when Strep A present for all but two cases subsequently resolved by lower concentration	N/A
Interfering Substances	No interference from common substances found in throat samples	100% agreement with expected results for most tested substances at specified concentrations	N/A

Study Details:

Sample Size and Data Provenance:
- Test Set (Clinical Study):
  - Evaluable for Accula vs. Culture: 654 samples from 669 enrolled subjects.
  - Evaluable for Accula vs. Molecular Comparator: 648 samples from 669 enrolled subjects.
  - Provenance: Prospective clinical study conducted at nine U.S. sites from May 2019 to January 2020.
- Reproducibility/Near-Cutoff Study: 90 samples per condition (Low Positive, Moderate Positive, Negative) tested across three CLIA-waived sites. These were contrived throat swabs.
- Limit of Detection (LoD): Replicates of 20 for confirmatory testing of two Strep A strains.
- Analytical Reactivity: 3 replicates per strain (4 strains total) at two concentrations.
- Analytical Specificity (Cross-Reactivity): 3 replicates per organism (47 organisms total), both in presence and absence of Strep A.
- Interfering Substances: 3 replicates per substance, positive and negative Strep A samples.
Number of Experts and Qualifications for Test Set Ground Truth:
- The document does not explicitly state the "number of experts" used to establish the ground truth for the clinical test set. However, for the reference methods:
  - Bacterial Culture (Blood Agar Culture): Performed at a "central laboratory" according to "instructions from the reference laboratory." This implies trained laboratory personnel, but specific qualifications are not detailed.
  - FDA-cleared molecular test (comparator): Performed at the central laboratory.
  - Second FDA-cleared molecular test (discrepant analysis): Used for all discrepant results.
Adjudication Method for the Test Set:
- For the clinical study, a discrepant analysis method was used. "All specimens generating discrepant results between the Accula Strep A Test and Blood Agar Culture, or between Accula and the molecular comparator test, were tested with a second FDA-cleared molecular test." This effectively acts as a "tie-breaker" or confirmatory method for unusual results.
Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study:
- No MRMC comparative effectiveness study was specifically described in terms of human readers' improvement with AI vs. without AI assistance.
- However, the Reproducibility/Near-Cutoff Study and the CLIA Waiver Studies involved "non-laboratory personnel" at "CLIA-waived sites" (Point of Care sites) to demonstrate that the device could be accurately used by intended users in the intended environment. This indirectly assesses the effectiveness of the device in the hands of typical users, rather than an AI assistance to human readers.
Standalone (Algorithm Only) Performance:
- Yes, the clinical performance described (Sensitivity, Specificity, PPA, NPA) represents the standalone performance of the Accula Strep A Test against established reference methods (Blood Agar Culture and FDA-cleared molecular tests). The device itself is a semi-automated system; these metrics evaluate its diagnostic accuracy independent of a human interpretation layer (beyond reading the visual result in the lateral flow).
Type of Ground Truth Used:
- For the clinical study, the primary ground truth was bacterial culture (Blood Agar Culture) for Streptococcus pyogenes.
- A second FDA-cleared molecular test was used as a confirmatory ground truth for discrepant results.
- Additionally, an FDA-cleared molecular comparator method served as another reference standard for direct comparison.
- For analytical studies (LoD, Reactivity, Specificity, Interfering Substances), the ground truth was established by contriving samples with known concentrations of specific organisms or substances.
Sample Size for the Training Set:
- The document describes performance studies (validation). It does not provide information on a "training set" in the context of machine learning, as this is a nucleic acid amplification test, not an AI-driven image analysis or algorithm that would typically require a training set. The device's components (enzymes, reagents, PCR technology) are developed and optimized rather than "trained."
How Ground Truth for the Training Set Was Established:
- Not applicable as described in item 7. The device operates on molecular principles and does not involve a machine learning training phase with a labeled dataset in the traditional sense.

Summary

{0}------------------------------------------------

Image /page/0/Picture/0 description: The image shows 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, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

November 9, 2020

Mesa Biotech, Inc. Barbara Stevens Regulatory Consultant 6190 Cornerstone Court, Suite 220 San Diego, California 92121

Re: K201269

Trade/Device Name: Accula Strep A Test Regulation Number: 21 CFR 866.2680 Regulation Name: Streptococcus Spp. Nucleic Acid-Based Assay Regulatory Class: Class II Product Code: PGX Dated: May 11, 2020 Received: May 12, 2020

Dear Barbara Stevens:

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.

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

{1}------------------------------------------------

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.

Ribhi Shawar, Ph.D. (ABMM) Chief. General Bacteriology and Antimicrobial Susceptibility Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

{2}------------------------------------------------

Section 5. 510(k) Summary of Safety and Effectiveness

This 510(k) summary of safety and effectiveness is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

The assigned 510(k) number is: _K201269

Sponsor/Applicant Name and Address 1. Mesa Biotech, Inc. Company Name: Address: 6190 Cornerstone Court, Suite 220 San Diego, CA 92121 Telephone: 858-800-4929 Contact Person: Barbara Stevens Regulatory Consultant Date Summary Prepared: 05/11/2020 Device Name and Classification 2. Trade Name: Accula™ Strep A Test Classification of Device: 21 CFR 866.2680, Streptococcus spp. Nucleic-acid based assay PGX Product Code

Predicate Device 3.

K141338, Roche cobas® Liat™ Strep A Test

Device Description 4.

Operating Principle

Strep A Kit Contents

The Accula Strep A Test Kit contains all the materials needed to run a test, except for the Accula Dock, which is provided separately. The Accula Strep A Test Kit contains the following components.

{3}------------------------------------------------

Sterile Swabs for throat swab collection (25) ●
Accula Strep A Buffer (25) ●
Accula Transfer Pipettes (25)
Accula Strep A Test Cassettes (25)
Strep A Positive Control Swab (1)
Negative Control Swab (1)
Instructions for Use
Quick Reference Guide

Indications for Use ട്.

The Accula Strep A Test performed on the Accula Dock is a molecular in vitro diagnostic test utilizing polymerase chain reaction (PCR) and lateral flow technologies for the qualitative, visual detection of Streptococcus pyogenes (Group A B-hemolytic Streptococcus, Strep A) bacterial nucleic acid. It is intended to aid in the rapid diagnosis of Group A Streptococcus bacterial infections from throat swabs of patients with signs and symptoms of pharyngitis.

Comparison to Predicate Device 6.

The following table provides a comparison of the characteristics of the Accula Strep A Test to the predicate device, the Roche cobas® Liat™ Strep A Test.

Item	510(k) Device:Mesa BiotechAccula Strep A Test	Predicate Device:Roche cobas® Liat™ Strep ATest (K141338)
Indications for Use	The Accula Strep A Testperformed on the AcculaDock is a molecular in vitro diagnostic testutilizing polymerasechain reaction (PCR)and lateral flowtechnologies for thequalitative, visualdetection ofStreptococcus pyogenes(Group A β-hemolyticStreptococcus , Strep A)bacterial nucleic acid. Itis intended to aid in therapid diagnosis of GroupA Streptococcus	The cobas® Strep A nucleicacid test for use on the cobasLiat System (cobas Strep A) isa qualitative in vitro diagnostictest for the detection ofStreptococcus pyogenes(Group A β-hemolyticStreptococcus , Strep A) inthroat swab specimens frompatients with signs andsymptoms of pharyngitis.The cobas Strep A assayutilizes nucleic acidpurification and polymerasechain reaction (PCR)
	bacterial infections fromthroat swabs of patientswith signs andsymptoms ofpharyngitis.All negative test results shouldbe confirmed by bacterialculture because negativeresults do not precludeinfection with Group AStreptococcus and should notbe used as the sole basis fortreatment.	technology to detectStreptococcus pyogenes bytargeting a segment of theStreptococcus pyogenesgenome.
Product Code	PGX	PGX
Analyte	Group A Streptococcus ( S. pyogenes )	Group A Streptococcus ( S. pyogenes )
Strep A Target	Conserved region of Group AStreptococcus genome	Conserved region of Group AStreptococcus genome
Sample Type	Throat swab	Throat swab
Assay Results	Qualitative	Qualitative
Intended Users andUse Locations	Clinical lab and CLIA-waivedsites	Clinical lab and CLIA-waivedsites
Bacterial Lysis/DNAExtraction	Detergent and heat	Chaotrope and enzymaticdigestion
Nucleic AcidPurification	No	Solid phase magnetic affinitycapture
Reagent Format	Unitized, ready for use	Unitized, ready for use
Internal Control	Yes	Yes
Positive andNegative ControlSwabs	Yes	Yes
Assay Technology	PCR amplification and visualidentification of amplificationproducts by hybridization to atest strip.	PCR nucleic acid amplificationand detection of specificamplification products usingmolecular TaqMan
		fluorescent probes.
Detection	Uses dyed microparticleconjugates to specificallydetect and identifyamplification reactionproducts.Visual interpretation of thepresence or the absence ofcolored lines on a test strip.	Uses fluorescently-labeledTaqman probe to specificallyidentify amplified cDNAproducts.Optical detection offluorescence
Instrument	Amplification controlled bythe Accula Dock.No detection by theinstrument.	Amplification and detectionperformed on the Liatinstrument.

{4}------------------------------------------------

{5}------------------------------------------------

7. Performance Summary

Expected Values

A total of 654 specimens were tested in a prospective clinical study conducted at nine U.S sites from May 2019 to January 2020. The overall prevalence of S. pyogenes as determined by bacterial culture was 20.0% (131/654), and as determined by the Accula Strep A Test, the overall positivity rate was 21.3% (139/654). The positivity rate by age range of the subjects is shown below.

	Strep A Positivity Rate by Accula Strep A Test
Age Group	Number of SwabSpecimens	Number of Strep APositives	Strep A Positivity Rate
≤ 5 Years	112	34	$34/112 = 30.36%$
6 to 21 Years	292	70	$70/292 = 23.97%$
22 to 59 Years	224	34	$34/224 = 15.18%$
≥ 60 Years	26	1	$1/26 = 3.85%$
Total	654	139	$139/654 = 21.25%$

Prospective Clinical Study:

Clinical performance characteristics of the Accula Strep A Test were evaluated in a multi-site prospective study from May 2019 to January 2020 in the U.S. A total of nine (9) investigational Point of Care sites participated in the study. To be enrolled in the study, patients had to be presenting at the participating study centers with symptoms of pharyngitis. Two throat swabs were collected from each subject using standard collection methods, a Copan FLOOTM swab for the Accula testing and a second swab using the Copan eSwab™ Liquid Amies Collection System

{6}------------------------------------------------

for the reference testing. One throat swab was tested, following elution in 2.5 mL of Accula Strep A Buffer, on the Accula Strep A Test, according to product instructions. The other throat swab was collected with the eSwab system in accordance with the instructions from the reference laboratory and the test manufacturer and transported to the central laboratory for testing by Blood Agar Culture and an FDA-cleared molecular test. All specimens generating discrepant results between the Accula Strep A Test and Blood Agar Culture, or between Accula and the molecular comparator test, were tested with a second FDA-cleared molecular test.

A total of 669 subjects were enrolled in this study. Of those, 15 samples were unevaluable for the Accula v. culture comparison due to sample transport and storage issues, failure to comply with inclusion/exclusion criteria, protocol deviations, and invalid results with the Accula test. A total of 654 samples were considered evaluable for this analysis. The performance of the Accula Strep A Test compared to Blood Agar Culture is shown in the table below. Results of the discrepant evaluations against both molecular methods are shown in the footnotes.

Mesa Biotech Accula™	Blood Agar Culture
Strep A Test	Positive	Negative	Total
Positive	126	13a	139
Negative	5b	510	515
Total	131	523	654
Sensitivity:	96.2% (126/131) (95% CI:91.4%-98.4%)
Specificity:	97.5% (510/523) (95% CI:95.8%-98.5%)
Accuracy:	97.2% (636/654) (95% CI:95.7%-98.3%)
NPV:	99% (510/515) (95% CI:97.7%-99.6%)
NPV (at 30% Prevalence):	98.3%

Accula Strep A Test Performance against Reference Culture

ªStrep A was detected in 7/13 False Positives specimens using the discrepant evaluation molecular method

*Strep A was not detected in 2/5 False Negative specimens using the discrepant evaluation molecular method

In addition, the Accula Strep A Test results were compared with an FDA-clear molecular comparative method. For this analysis, 21 of the samples from the 669 enrolled subjects were unevaluable due to sample transport and storage issues, failure to comply with inclusion/exclusion criteria, protocol deviations, invalid results with the molecular comparator method, and invalid results with the Accula test. A total of 648 samples were considered evaluable for this analysis. The performance of the Accula Strep A Test compared to the FDAcleared molecular method are shown in the table below. Results of the discrepant evaluation against the discrepant analysis molecular method are shown in the footnotes.

Accula™ Strep A Test Strep A Performance Against the Molecular Comparator

Mesa Biotech Accula™Strep A Test	Molecular Comparator
	Positive	Negative	Total
Positive	137	1a	138
Negative	9b	501	510
Total	146	502	648

{7}------------------------------------------------

PPA (Positive Percent
Agreement):	93.8% (137/146) (95% CI:88.7%-96.7%)
NPA (Negative
Percent Agreement):	99.8% (501/502) (95% CI:98.9%-100%)

a Strep A was detected in 0/1 False Positives specimens using the discrepant evaluation molecular method

b Strep A was not detected in 5/9 False Negative specimens using the discrepant evaluation molecular method

Reproducibility/Near-Cutoff Study

The Reproducibility study was performed to demonstrate the reproducibility of the Accula Strep A Test with contrived throat swabs, including samples at analyte concentrations near the assay cutoff, at three CLIA waived sites that also participated in the clinical study. The objective of the study was to demonstrate reproducibility of the assay in the hands of multiple users at multiple sites over multiple non-consecutive days.

The test panel consisted of three samples at varying Strep A concentrations, two of which are at or near the assay cutoff. Each positive sample was prepared by spiking the Streptococcus pyogenes strain BAA-946 into clinical matrix. The targeted concentration for the Low Positive Sample was 1x LoD and for the Moderate Positive Sample was 2x LoD. The Negative Sample contained no Strep A.

Samples were blinded and coded and were provided to testing operators in a random fashion. Testing was performed with triplicate swab preparations, by two operators per site, on five nonconsecutive days over a period of two weeks, concurrently with the clinical study.

Results are reported as percent: observed result/expected result x 100. No significant differences were observed within run, between runs, between operators or between sites. Agreement by site is summarized in the table below.

	Site
	1		2		3		Overall
SampleType	PercentAgreement	Count	PercentAgreement	Count	PercentAgreement	Count	PercentAgreement(95% CI)
LowPositive	96.7%	29/30	100.0%	30/30	100.0%	30/30	98.9% (89/90)(94.0%-99.8%)
ModeratePositive	96.7%	29/30	100.0%	30/30	96.6%	28/29	97.8% (87/89)(92.2%-99.4%)
Negative	100.0%	30/30	100.0%	30/30	93.3%	28/30	97.8% (88/90)(92.3%-99.4%)

Site to Site Reproducibility: Percent Agreement and Total Counts (Observed/Expected)

Limit of Detection

{8}------------------------------------------------

Two Strep A strains were tested at multiple analyte levels until the LoD was determined (the level at which at least 19/20 results are positive). Bacteria were serially diluted into a pooled negative clinical matrix and spiked onto a swab for each replicate to create the contrived test samples. Confirmatory testing was performed in replicates of twenty (20) on multiple days. The results are summarized in the table below.

Strep A Strain	LoD Level1
BAA-946	75 CFU/mL
ATCC 19615	10 CFU/mL

Accula Strep A Limit of Detection

1 Final concentration of organisms after 10 uL of bacterial dilution is spiked onto swab and eluted in 2.5 mL Strep A Buffer (assuming 100% recovery of bacteria).

Analytical Reactivity

Inclusivity verification was evaluated for the Accula Strep A Test at Mesa Biotech. The panel consisted of four (4) additional Strep A strains that were not included in the LoD study. Each strain was tested in triplicate at concentrations of approximately 1.5x LoD and 3.0x LoD. Test results are summarized in the table below. All strains were detected at the levels tested.

Inclusivity Results by Strain

Strep A Strain	1.5X LoDConcentration	Percent Detection(# Positive/3)	3.0X LoDConcentration	Percent Detection(# Positive/3)
ATCC 10403	112.5 CFU/mL	100% (3/3)	225 CFU/mL	100% (3/3)
ATCC 21548	112.5 CFU/mL	66.67% (2/3)	225 CFU/mL	100% (3/3)
ATCC 700294	112.5 CFU/mL	100% (3/3)	225 CFU/mL	100% (3/3)
ATCC 700497	112.5 CFU/mL	100% (3/3)	225 CFU/mL	100% (3/3)

The Analytical Specificity (Cross-Reactivity)

Cross-reactivity was evaluated by testing 47 potentially cross-reacting organisms with the Accula Strep A Test. Each organism was diluted in a clinical matrix, both in the absence and in the presence of 3x LoD Strep A, and tested in triplicate. The organisms, concentrations, and test results are shown in the tables below. All 47 organisms gave negative results when tested in the absence of 3x LoD Strep A. None of the 47 organisms interfered with the detection of Strep A when tested in the presence of 3x LoD Strep A.

Testing of Potential Cross-Reactive Organisms in the Absence of Strep A

OrganismKey #	Organism Name	Test Level	Test Results (# ofStrep A Pos /3)
1	Adenovirus Type 1	1.00E+06 TCID50/mL	0/3
2	Arcanobacteriumhaemolyticum	2.00E+07 CFU/ml	0/3
3	Bacillus cereus	1.00E+06 CFU/ml	0/3
4	Bordetella pertussis	2.00E+06 CFU/ml	0/3
5	Burkholderia cepacia	2.00E+07 CFU/ml	0/3
OrganismKey #	Organism Name	Test Level	Test Results (# ofStrep A Pos /3)
6	Campylobacter rectus	1.00E+06 CFU/ml	0/3
7	Candida albicans	2.00E+06 CFU/ml	0/3
8	Corynebacterium diphtheriae	5.00E+06 CFU/ml	0/3
9	Enterococcus faecalis	2.00E+06 CFU/ml	0/3
10	Escherichia coli	2.00E+07 CFU/ml	0/3
11	Fusobacterium necrophorum	1.00E+06 CFU/ml	0/3
12	Haemophilus influenzae	1.00E+06 CFU/ml	0/3
13	Human Influenza virus A****	1.00E+06 TCID50/mL	0/3
14	Human Influenza virus B	1.00E+06 TCID50/mL	0/3
15	Human metapneumovirus	1.00E+06 TCID50/mL	0/3
16	Klebsiella pneumoniae	2.00E+07 CFU/ml	0/3
17	Lactobacillus acidophilus	2.00E+06 CFU/ml	0/3
18	Lactococcus lactis	2.00E+07 CFU/ml	0/3
19	Legionella longbeachae	1.00E+07 CFU/ml	0/3
20	Moraxella catarrhalis	1.00E+06 CFU/ml	0/3
21	Mycoplasma pneumoniae	1.00E+06 CCU/ml	0/3
22	Neisseria gonorrhoeae	3.00E+06 CFU/ml	0/3
23	Parainfluenza Type 3	1.00E+06 TCID50/mL	0/3
24	Parvimonas micra(Peptostreptococcus micros)	1.50E+06 CFU/ml	0/3
25	Prevotella oralis (Bacteroidesoralis)	1.00E+06 CFU/ml	0/3
26	Pseudomonas aeruginosa	1.00E+06 CFU/ml	0/3
27	Respiratory syncytial virusType B	1.00E+06 TCID50/mL	0/3
28	Rhinovirus	1.00E+06 TCID50/mL	0/3
29	Saccharomyces cerevisiae	2.00E+06 CFU/ml	0/3
30	Staphylococcus epidermidis	5.00E+07 CFU/ml	0/3
31	Stenotrophomonasmaltophilia	5.00E+07 CFU/ml	0/3
32	Streptococcus agalactiae	2.00E+06 CFU/ml	0/3
33	Streptococcus anginosus	2.00E+06 CFU/ml	0/3
34	Streptococcus bovis	5.00E+06 CFU/ml	0/3
35	Streptococcus canis	2.00E+07 CFU/ml	0/3
36	Streptococcus constellatussubsp. Pharyngis	3.00E+06 CFU/ml	0/3
37	Streptococcus dysgalactiaesubsp. Equisimilis	1.00E+06 CFU/ml	0/3
38	Streptococcus gallolyticus	2.00E+06 CFU/ml	0/3
39	Streptococcus intermedius	2.00E+06 CFU/ml	0/3
40	Streptococcus mitis	2.00E+06 CFU/ml	0/3
OrganismKey #	Organism Name	Test Level	Test Results (# ofStrep A Pos /3)
41	Streptococcus mutans	2.00E+07 CFU/ml	0/3
42	Streptococcus oralis	2.00E+06 CFU/ml	0/3
43	Streptococcus pneumonia	2.00E+06 CFU/ml	0/3
44	Streptococcus salivarius	2.00E+06 CFU/ml	0/3
45	Streptococcus sanguinus	2.00E+06 CFU/ml	0/3
46	Treponema denticola	2.00E+06 CFU/ml	0/3
47	Veillonella parvula	2.00E+07 CFU/ml	0/3

{9}------------------------------------------------

{10}------------------------------------------------

Testing of Potential Cross-Reactive Organisms in the Presence of 3x LoD Strep A

Organism Key #	Organism Name	Test Level	Test Results (# ofStrep A Pos /3)
1	Adenovirus Type 1	1.00E+06 TCID50/mL	3/3
2	Arcanobacteriumhaemolyticum	2.00E+07 CFU/ml	3/3
3	Bacillus cereus	1.00E+06 CFU/ml	3/3
4	Bordetella pertussis	2.00E+06 CFU/ml	3/3
5	Burkholderia cepacia	2.00E+07 CFU/ml	3/3
6	Campylobacter rectus	1.00E+06 CFU/ml	3/3
7	Candida albicans	2.00E+06 CFU/ml	3/3
8	Corynebacterium diphtheriae	5.00E+06 CFU/ml	3/3
9	Enterococcus faecalis	2.00E+06 CFU/ml	3/3
10	Escherichia coli 1	1.00E+07 CFU/ml	3/3
11	Fusobacterium necrophorum	1.00E+06 CFU/ml	3/3
12	Haemophilus influenzae	1.00E+06 CFU/ml	3/3
13	Human Influenza virus A	1.00E+06 TCID50/mL	3/3
14	Human Influenza virus B	1.00E+06 TCID50/mL	3/3
15	Human metapneumovirus	1.00E+06 TCID50/mL	3/3
16	Klebsiella pneumoniae	2.00E+07 CFU/ml	3/3
17	Lactobacillus acidophilus	2.00E+06 CFU/ml	3/3
18	Lactococcus lactis	2.00E+07 CFU/ml	3/3
19	Legionella longbeachae	1.00E+07 CFU/ml	3/3
20	Moraxella catarrhalis	1.00E+06 CFU/ml	3/3
21	Mycoplasma pneumoniae	1.00E+06 CCU/ml	3/3
22	Neisseria gonorrhoeae	3.00E+06 CFU/ml	3/3
23	Parainfluenza Type 3	1.00E+06 TCID50/mL	3/3
24	Parvimonas micra(Peptostreptococcus micros)	1.50E+06 CFU/ml	3/3

25	Prevotella oralis (Bacteroidesoralis)	1.00E+06 CFU/ml	3/3

26	Pseudomonas aeruginosa	1.00E+06 CFU/ml	3/3

{11}------------------------------------------------

OrganismKey #	Organism Name	Test Level	Test Results (# ofStrep A Pos /3)
27	Respiratory syncytial virusType B	1.00E+06 TCID50/mL	3/3
28	Rhinovirus 2	5.00E+05 TCID50/mL	3/3
29	Saccharomyces cerevisiae	2.00E+06 CFU/ml	3/3
30	Staphylococcus epidermidis	5.00E+07 CFU/ml	3/3
31	Stenotrophomonasmaltophilia	5.00E+07 CFU/ml	3/3
32	Streptococcus agalactiae	2.00E+06 CFU/ml	3/3
33	Streptococcus anginosus	2.00E+06 CFU/ml	3/3
34	Streptococcus bovis	5.00E+06 CFU/ml	3/3
35	Streptococcus canis	2.00E+07 CFU/ml	3/3
36	Streptococcus constellatussubsp. pharyngis	3.00E+06 CFU/ml	3/3
37	Streptococcus dysgalactiaesubsp. equisimilis	1.00E+06 CFU/ml	3/3
38	Streptococcus gallolyticus	2.00E+06 CFU/ml	3/3
39	Streptococcus intermedius	2.00E+06 CFU/ml	3/3
40	Streptococcus mitis	2.00E+06 CFU/ml	3/3
41	Streptococcus mutans	2.00E+07 CFU/ml	3/3
42	Streptococcus oralis	2.00E+06 CFU/ml	3/3
43	Streptococcus pneumonia	2.00E+06 CFU/ml	3/3
44	Streptococcus salivarius	2.00E+06 CFU/ml	3/3
45	Streptococcus sanguinus	2.00E+06 CFU/ml	3/3
46	Treponema denticola	2.00E+06 CFU/ml	3/3
47	Veillonella parvula	2.00E+07 CFU/ml	3/3

1 Escherichia coli was originally tested at 2 x 107 CFU/mL and gave 2/3 positive results in the presence of Strep A. No cross-reactivity was observed at 1 x 107 CFU/mL.

2 Rhinovirus was tested at 1 x 106 TCID50/mL and gave the expected 3/3 in the presence of Strep A. However, the visually graded maximum line intensity of "3" was not observed. No interference with line intensity was observed at a concentration of 5 x 105 TCID50/mL.

Interfering Substances

To assess substances with the potential to interfere with the performance of the Accula Strep A Test, samples with and without Strep A were tested in replicates of three (3) with each interfering substance at "worst case" concentrations, in addition to a "no interferent" control sample. Positive Strep A contrived samples were prepared by spiking the Strep A strain BAA-946 into a clinical throat swab matrix. The Negative sample was the clinical throat swab matrix alone. Any potential interferents that showed inhibition in the Accula Strep A Test were diluted and re-tested at a lower concentration. The table below summarizes the interferents tested, the highest concentration that showed no inhibition, the samples tested and the test results.

Effect of Potentially Interfering Substances on Accula Strep A Test Performance

{12}------------------------------------------------

Potential Interferent	Active Ingredient	FinalConcentration	Target	% Agreementwith ExpectedResults
Blood (Human) 1	NA	50% (v/v)12.5% (v/v)	Strep AIC	100% (3/3)
Chloroseptic Max	Phenol 1.5%,Glycerin 33%	100% (v/v)	Strep AIC	100% (3/3)
Cold&Flu Relief CoughSyrup	Acetaminophen21.7 mg/mL,Dextromethorphan0.67 mg/mL,Guaifenesin 13.3mg/mL,Phenylephrine 0.33mg/mL	100% (v/v)	Strep AIC	100% (3/3)
Listerine Cool MintAntiseptic Mouth Wash	Eucalyptol 0.092%,Menthol 0.042%,Methyl Salicylate0.060%, Thymol0.064%	100% (v/v)	Strep AIC	100% (3/3)
Cepacol (throat lozenge)	Benzocaine,Menthol	0.3% Benzocaine(w/v), 0.046%Menthol (w/v)	Strep AIC	100% (3/3)
Sucrets	DyclonineHydrochloride,Menthol	0.06%DyclonineHydrochloride(w/v), 0.12%Menthol (w/v)	Strep AIC	100% (3/3)
Crest Pro HealthFluoride Toothpaste	Stannous Fluoride0.454% (0.14%W/V Fluoride Ion)	100% (v/v)	Strep AIC	100% (3/3)
Halls Triple SoothingCough Drops*	Eucalyptus Oil	100% (v/v)	Strep AIC	100% (3/3)
Advil Liqui-Gels	Ibuprofen	100% (v/v)	Strep AIC	100% (3/3)
Miralax	PolyethyleneGlycol	30.4% (w/v)	Strep AIC	100% (3/3)
Tums Extra Strength 2	Calcium Carbonate	20 mg/mL30 mg/mL	Strep AIC	100% (3/3)
Food Dye	N/A	100% (v/v)	Strep AIC	100% (3/3)
Whole Milk (Dairy) 3	N/A	12.50% (v/v)50.00% (v/v)	Strep AIC	100% (3/3)
Orange Juice 4	N/A	50% (v/v)100% (v/v)	Strep AIC	100% (3/3)
Penicillin G	Penicillin GSodium Salt	100 mg/mL	Strep AIC	100% (3/3)
Potential Interferent	Active Ingredient	FinalConcentration	Target	% Agreementwith ExpectedResults
Cephalexin	Cephalexin	25 mg/mL	Strep A	100% (3/3)
Cephalexin	Cephalexin	25 mg/mL	IC	100% (3/3)
Mucin, Type II (fromporcine stomach)	Purified mucinprotein	50 mg/mL	Strep A	100% (3/3)
Mucin, Type II (fromporcine stomach)	Purified mucinprotein	100 mg/mL	IC	100% (3/3)
Tobramycin(antibacterial)	Tobramycin	75 mg/mL	Strep A	100% (3/3)
Tobramycin(antibacterial)	Tobramycin	75 mg/mL	IC	100% (3/3)
Amoxicillin	Amoxicillin	100 mg/mL	Strep A	100% (3/3)
Amoxicillin	Amoxicillin	100 mg/mL	IC	100% (3/3)
No interferent	N/A	N/A	Strep A	100% (3/3)
No interferent	N/A	N/A	IC	100% (3/3)

{13}------------------------------------------------

IC: Internal Control, for negative samples

*Eucalyptus oil, an active ingredient in Halls cough drops, was used in place of Halls Triple Soothing

Cough Drops.

1 Human blood showed inhibition at 100% concentration for Strep A detection in the positive sample, but no inhibition at 50% concentration. Human blood showed inhibition at 100%. 50% and 25% concentrations for IC detection in the negative sample, but no inhibition at 12.5% concentration.
2 Tums was initially tested with a solution of 1.5 g/mL (1 Tum dissolved into 2.5 mL of Strep A Buffer). This inhibited all reactions. The Strep A Positive sample was inhibited when tested with an additional 8x and 32x dilution. No inhibition was observed at a concentration of 20 mg/mL for the Strep A Positive sample. The Negative Strep A sample showed inhibition when tested with an additional 8x dilution. No inhibition was observed at a concentration of 30 mg/mL for the Strep A Negative sample.
3 Milk showed inhibition at 100%, 50% and 25% concentrations for Strep A detection in the positive sample, but no inhibition at 12.5% concentration. Milk showed inhibition at the 100% concentration for IC detection in the negative sample, but no inhibition at 50% concentration.
4 Orange juice showed inhibition at 100% concentration for detection of Strep A in the positive sample, but no inhibition at 50%.

CLIA Waiver Studies

Comparison with a Reference Method:

The performance of the Accula Strep A Test was evaluated at nine Point of Care sites by nonlaboratory personnel in a prospective clinical study from May 2019 to January 2020 in the U.S. Throat swabs were collected from patients with symptoms of pharyngitis and were tested with the Accula Strep A Test, the Blood Agar Culture reference method, and an FDA-cleared molecular comparator method. All specimens generating discrepant results were evaluated using a second molecular comparative method.

{14}------------------------------------------------

The comparison with culture showed a sensitivity of 96.2% (95% confidence interval: 91.4%-98.4%) and a specificity of 97.5% (95% CI: 95.8%-98.5%). The comparison with the molecular comparator method showed Positive Percent Agreement of 93.8% (95% CI: 88.7%-96.7%) and Negative Percent Agreement of 99.8% (95% CI: 98.9%-100%). The study demonstrates that non-laboratory personnel in an intended use environment can obtain results with the Accula Strep A test equivalent to results for the comparator methods.

Test Performance near the Assay Cutoff

Three CLIA-waived sites that participated in the prospective clinical study also participated in the Reproducibility Study. Two non-laboratory operators per site tested contrived throat swab samples at and above the assay cutoff, in addition to a negative sample, on five non-consecutive days. Results showed good agreement of observed test results with expected results. For the Low Positive sample (1x LoD), agreement was 98.9% (95% CI: 94.0%-99.8%). For the Moderate Positive sample (2x LoD), agreement was 97.8% (95% CI: 92.2%-99.4%). For the Negative sample, agreement was 97.8% (95% CI: 92.3%-99.4%).

The study demonstrates that non-laboratory personnel in CLIA waived settings can achieve accurate results when testing samples at or near the assay cutoff.

8. Conclusion

The information presented in this Premarket Notification demonstrates that the performance of the Accula Strep A Test is substantially equivalent in intended use, technological characteristics, and performance to the predicate device, thereby supporting 510(k) clearance.

Regulation Number and Section

§ 866.2680

Streptococcus spp. nucleic acid-based assay.(a)
Identification. AStreptococcus spp. nucleic acid-based assay is a qualitative in vitro diagnostic device intended to simultaneously detect and identify variousStreptococcus spp. nucleic acids extracted directly from clinical specimens. The device detects specific nucleic acid sequences for organism identification. The identification aids in the diagnosis of diseases caused by bacteria belonging to the genusStreptococcus and provides epidemiological information on these diseases. Pathogenic streptococci are associated with infections, such as sore throat, impetigo (an infection characterized by small pustules on the skin), urinary tract infections, rheumatic fever, and kidney disease.(b)
Classification. Class II (special controls). The special controls for this device are:(1) Premarket notification submissions must include detailed device description documentation, including the device components, ancillary reagents required but not provided, and a detailed explanation of the methodology including primer/probe sequence, design, and rationale for sequence selection.
(2) Premarket notification submissions must include detailed documentation from the following analytical and clinical performance studies: Analytical sensitivity (Limit of Detection), reactivity, inclusivity, precision, reproducibility, interference, cross reactivity, carry-over, and cross contamination.
(3) Premarket notification submissions must include detailed documentation from a clinical study. The study, performed on a study population consistent with the intended use population, must compare the device performance to results obtained from well-accepted reference methods.
(4) Premarket notification submissions must include detailed documentation for device software, including, but not limited to, software applications and hardware-based devices that incorporate software.
(5) Premarket notification submissions must include database implementation methodology, construction parameters, and quality assurance protocols, as appropriate.
(6) The device labeling must include limitations regarding the need for culture confirmation of negative specimens, as appropriate.
(7) A detailed explanation of the interpretation of results and acceptance criteria must be included in the device's 21 CFR 809.10(b)(9) compliant labeling.
(8) Premarket notification submissions must include details on an end user device training program that will be offered while marketing the device, as appropriate.