Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description and performance studies focus on real-time PCR technology for the qualitative detection of bacterial DNA. There is no mention of AI or ML in the document.

Therapeutic

No
The device is an in vitro diagnostic (IVD) assay intended for the qualitative detection of Group A Streptococcus to aid in diagnosis, not to provide therapy.

Diagnostic

Yes
The "Intended Use / Indications for Use" section explicitly states, "This test is intended for use as an aid in the diagnosis of GAS infection."

SaMD (Software as a Medical Device)

No

The device description explicitly states that the system consists of the assay, the 3M Integrated Cycler (a hardware component), the Direct Amplification Disc (DAD), and associated accessories, in addition to the software.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the Simplexa™ Group A Strep Direct assay is intended for "in vitro qualitative detection of Group A Streptococcus (GAS) from throat swabs collected from human patients". The term "in vitro" means "in glass" or "in the laboratory," referring to tests performed outside of the living organism.
Purpose: The assay is used to detect the presence of Group A Streptococcus in a patient sample (throat swab) to aid in the diagnosis of GAS infection. This is a classic function of an in vitro diagnostic device.
Device Description: The description details a laboratory-based system (real-time PCR) that analyzes a biological sample (throat swab) to detect specific bacterial DNA.
Control Pack: The Simplexa™ Group A Strep Positive Control Pack is also described as being used "as a control with Simplexa™ Group A Strep Direct," further indicating its use in a laboratory testing setting.
Intended User / Care Setting: The specified settings (hospital, reference, or state laboratory) are typical environments where IVD tests are performed.

All of these points align with the definition and characteristics of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

Simplexa™ Group A Strep Direct

The Focus Diagnostics Simplexa™ Group A Strep Direct assay is intended for use on the 3M Integrated Cycler for the in vitro qualitative detection of Group A Streptococcus (GAS) from throat swabs collected from human patients with signs and symptoms of pharyngitis, such as sore throat. This test is intended for use as an aid in the diagnosis of GAS infection. The assay is intended for use in hospital, reference, or state laboratory settings. The device is not intended for point-of-care use.

Simplexa™ Group A Strep Positive Control Pack

Focus Diagnostics' Simplexa™ Group A Strep Positive Control Pack is intended to be used as a control with Simplexa™ Group A Strep Direct. This control is not intended for use with other assays or systems.

Product codes (comma separated list FDA assigned to the subject device)

PGX, OOI

Device Description

The Simplexa™ Group A Strep Direct assay system is a real-time PCR system that enables the direct amplification and qualitative detection of Group A Strep bacterial DNA from throat swabs that have not undergone a nucleic acid extraction. The system consists of the Simplexa™ Group A Strep Direct assay, the 3M Integrated Cycler (with Integrated Cycler Studio Software), the Direct Amplification Disc (DAD) and associated accessories.

In the Simplexa™ Group A Strep Direct assay, bi-functional fluorescent probe-primers are used together with corresponding reverse primers to amplify Group A Strep bacterial DNA and the Internal Control (DNA IC). The assay targets a conserved region of Group A Strep (pyroqenic exotoxin B gene) to identify this bacteria in the specimen. The DNA IC is used to detect PCR failure and/or inhibition.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Throat swabs, pharynx

Indicated Patient Age Range

Not Found

Intended User / Care Setting

hospital, reference, or state laboratory settings

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

Not Found

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

REPRODUCIBILITY

Three (3) investigative sites assessed the device's inter-site, inter-day and interlintra-assay reproducibility. Each of the laboratories tested a panel of six (6) members that included contrived samples at low positive, (approximately 1 X LoD) and moderately positive sample (approximately 3 X LoD) for M1 and M3 serotypes of Group A Streptococus, a positive and negative control were also included in the panel. The assays were performed in triplicate (3) on five (5) different days. Each site had two (2) operators who each assayed the sample panel once (1) per day, for a total of two (2) sets of data per day. Combined results for all sites are presented in the tables below.
Overall % Agreement with Expected Results: 96.9% (523/540) (95% CI: 95.0% to 98.0%)

ANALYTICAL SENSITIVITY/LIMIT OF DETECTION

The Limit of Detection (LoD) was determined for the Simplexa™ Group A Strep Direct by performing a dilution series. The LoD samples used for the study were contrived using two (2) serotypes of Group A Streptococcus; M1 and M3 that were verified (re-grown and re-titered) bacterial stock. For each serotype, eight (8) different concentrations were spiked in simulated matrix from the verified bacterial and confirmed using thirty two (32) replicates.

LoD:
M1: 682 cfu/mL
M3: 2350 cfu/mL

ANALYTICAL REACTIVITY / CROSS REACTIVITY

Analytical Reactivity was assessed for the ability of Simplexa™ Group A Strep Direct to detect sixty (60) Group A Streptococus strains not present in the LoD study. Analytical Reactivity was observed in testing of twenty one (21) strains. All were detected as positive for Group A Strep at or below 5000 cfu/mL. Analytical Reactivity was tested using in silico NCBI BLAST sequence analysis methods for thirty nine (39) GAS strains distinct from the wet tested strains.

Analytical reactivity was evaluated for the Simplexa™ Group A Strep the ability to exclusively identify Group A Streptococcus with no cross reactivity to organisms that are closely related, or cause similar clinical symptoms, or present as normal flora in the pharynx. Negative specimens were spiked with potentially cross-reactive organisms at known concentrations. Spiked specimens were examined for reactivity with the Group A Strep Direct. Sixty four (64) organisms were tested. Bacteroides ovalis and Tremella fuciformis were not available for testing therefore data from In-Silico NCBI BLAST sequence analysis was performed. No cross reactivity was found.

INTERFERENCE

The performance of Simplexa™ Group A Strep Direct was evaluated with potentially interfering substances that may be present in pharynx. The potentially interfering substances were evaluated in a contrived sample that contained Group A Streptococus at low positive concentrations (approximately 1 X LoD) and moderate positive concentrations (approximately 3 X LoD) of Group A Streptococus serotypes M1 and M3. There was no evidence of interference caused by the substances at the concentrations tested.

CLINICAL PROSPECTIVE STUDY

One thousand three hundred and ninety seven (1397) samples that were prospectively collected from four (4) geographically diverse sites between May 6, 2014 from patients with signs and symptoms of Group A Strep infections of the pharynx. One thousand three hundred and fifty two (1352) samples were evaluable on Simplexa™ Group A Strep Direct and the comparator culture method. Samples were tested on Simplexa™ Group A Strep Direct at the collection sites and comparator culture method was performed at one (1) central laboratory. The invalid rate of the clinical prospective study using Simplexa™ Group A Strep Direct was 0.57% (eight out of one thousand three hundred and ninety six 8/1396 samples). Discrepant analysis was performed using a validated bidirectional sequencing assay.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Sensitivity: 97.4% (152/156) (95% CI: 93.6% to 99.0%)
Specificity: 95.2% (1139/1196) (95% CI: 93.9% to 96.3%)
Positive Predictive Value (PPV): 72.7% (152/209) (95% CI: 66.3% to 78.3%)
Negative Predictive Value (NPV): 99.7% (1139/1143) (95% CI: 99.1% to 99.9%)

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

Lyra™ Direct Strep Assay K133883

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

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.

Summary

0

Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized depiction of an eagle or bird with three human profiles incorporated into its design. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular fashion around the emblem.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

FOCUS DIAGNOSTICS, INC. SHARON YOUNG SENIOR REGULATORY AFFAIRS SPECIALIST 11331 VALLEY VIEW ST CYPRESS CA 90630

March 18. 2015

Re: K143651

Trade/Device Name: Simplexa™ Group A Strep Direct, Simplexa™ Group A Strep Positive Control Pack Regulation Number: 21 CFR 866.2680 Regulation Name: Streptococcus spp. nucleic acid-based assay Regulatory Class: II Product Code: PGX, OOI Dated: December 22, 2014 Received: December 23, 2014

Dear Ms. Young:

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. 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 Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

1

If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. 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

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Uwe Scherf -S for

Sally A. Hojvat, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

2

Indications for Use

510(k) Number (if known) K143651

Device Name

Simplexa™ Group A Strep Direct and Simplexa™ Group A Strep Positive Control Pack

Indications for Use (Describe)

SimplexaTM Group A Strep Direct

The Focus Diagnostics Simplexa™ Group A Strep Direct assay is intended for use on the 3M Integrated Cycler for the in vitro qualitative detection of Group A Streptococcus (GAS) from throat swabs collected from human patients with signs and symptoms of pharyngitis, such as sore throat. This test is intended for use as an aid in the diagnosis of GAS infection. The assay is intended for use in hospital, reference, or state laboratory settings. The device is not intended for point-ofcare use.

Simplexa™ Group A Strep Positive Control Pack

Focus Diagnostics' Simplexa™ Group A Strep Positive Control Pack is intended to be used as a control with Simplexa™ Group A Strep Direct. This control is not intended for use with other assays or systems.

			Type of Use (Select one or both, as applicable)
--	--	--	-------------------------------------------------

X Prescription Use (Part 21 CFR 801 Subpart D)

Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

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

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Image /page/3/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo features the word "FOCUS" in bold, sans-serif font. A curved, black shape is positioned to the left of the word, resembling a bracket or a stylized eye. Below "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin, horizontal line.

510(k) Summary

Simplexa™ Group A Strep Direct Catalog No. MOL2850 Simplexa™ Group A Strep Positive Control Pack Catalog No. MOL2860 March 17, 2015 Page 1 of 10

| Applicant | Focus Diagnostics, Inc.
11331 Valley View Street
Cypress, California 90630
USA |
|----------------------------------------------|-----------------------------------------------------------------------------------------|
| Establishment Registration No. | 2023365 |
| Contact Person | Sharon Young
tel 562.240.6680
fax 562.240.6529
syoung@focusdx.com |
| Summary Date | March 17, 2015 |
| Proprietary Name | Simplexa™ Group A Strep Direct
Simplexa™ Group A Strep Positive Control Pack |
| Generic Name | Group A Streptococcus nucleic acid |
| Classification
Regulation
Product Code | Class II
21 CFR 866.2680
PGX |
| Predicate Device | Lyra™ Direct Strep Assay K133883, |

Intended Use

Simplexa™ Group A Strep Direct

The Focus Diagnostics Simplexa™ Group A Strep Direct assay is intended for use on the 3M Integrated Cycler for the in vitro qualitative detection of Group A Streptococcus (GAS) from throat swabs collected from human patients with signs and symptoms of pharvnaitis, such as sore throat. This test is intended for use as an aid in the diagnosis of GAS infection. The assay is intended for use in hospital, reference, or state laboratory settings. The device is not intended for point-of-care use.

Simplexa™ Group A Strep Positive Control Pack

Focus Diagnostics' Simplexa™ Group A Strep Positive Control Pack is intended to be used as a control with Simplexa™ Group A Strep Direct. This control is not intended for use with other assays or systems.

Device Description

The Simplexa™ Group A Strep Direct assay system is a real-time PCR system that enables the direct amplification and qualitative detection of Group A Strep bacterial DNA from throat swabs that have not undergone a nucleic acid extraction. The system consists of the Simplexa™ Group A Strep Direct assay, the 3M Integrated Cycler (with Integrated Cycler Studio Software), the Direct Amplification Disc (DAD) and associated accessories.

In the Simplexa™ Group A Strep Direct assay, bi-functional fluorescent probe-primers are used together with corresponding reverse primers to amplify Group A Strep bacterial DNA and the Internal Control (DNA IC). The assay targets a conserved region of Group A Strep (pyroqenic exotoxin B gene) to identify this bacteria in the specimen. The DNA IC is used to detect PCR failure and/or inhibition.

4

Image /page/4/Picture/0 description: The image shows the logo for Focus Diagnostics. The word "FOCUS" is in large, bold, black letters. There is a curved, black shape to the left of the word. Below the word "FOCUS" is the word "Diagnostics" in smaller, black letters with a line above it.

510(k) Summary

Simplexa™ Group A Strep Direct Catalog No. MOL2850 Simplexa™ Group A Strep Positive Control Pack Catalog No. MOL2860 March 17, 2015 Page 2 of 10

Predicate Device Information

Item	Device	Predicate
Name	Simplexa™ Group A Strep Direct	Lyra™ Direct Strep Assay
K133883
Intended Use	Simplexa™ Group A Strep Direct
The Focus Diagnostics Simplexa™ Group A Strep Direct assay is intended for use on the 3M Integrated Cycler for the in vitro
qualitative detection of Group A
Streptococcus (GAS) from throat swabs
collected from human patients with signs and
symptoms of pharyngitis, such as sore throat.
This test is intended for use as an aid in the
diagnosis of GAS infection. This test is
intended for use as an aid in the diagnosis of
GAS infection. The assay is intended for use
in hospital, reference, or state laboratory
settings. The device is not intended for point-of-care use.

Simplexa™ Group A Strep Positive Control Pack
Focus Diagnostics' Simplexa™ Group A
Strep Positive Control Pack is intended to be
used as a control with Simplexa™ Group A
Strep Direct. This control is not intended for
use with other assays or systems. | The Lyra Direct Strep Assay is a Real-Time PCR in vitro diagnostic test for the qualitative detection and differentiation of Group A β-hemolytic Streptococcus (Streptococcus pyogenes) and pyogenic Group C and G β-hemolytic Streptococcus nucleic acids isolated from throat swab specimens obtained from patients with signs and symptoms of pharyngitis, such as a sore throat. The assay does not differentiate between pyogenic Groups C and G β-hemolytic Streptococcus.

All negative test results should be confirmed by bacterial culture, because negative results do not preclude Group A, C or G Strep infection and should not be used as the sole basis for treatment.

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Image /page/5/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of a stylized black swoosh on the left, followed by the word "FOCUS" in bold, black capital letters. Below "FOCUS" is the word "Diagnostics" in a smaller, non-bold font, underlined with a thin black line.

510(k) Summary

Simplexa™ Group A Strep Direct Catalog No. MOL2850 Simplexa™ Group A Strep Positive Control Pack Catalog No. MOL2860 March 17, 2015 Page 3 of 10

ltem	Device	Predicate
Name	Simplexa™ Group A Strep Direct	Lyra™ Direct Strep Assay
K133883
Performance
Characteristics	Sensitivity: 97.4% (152/156) (95% Cl: 93.6%
to 99.0%)
Specificity: 95.2% (1139/1196) (95% Cl:
93.9% to 96.3%)	GAS Sensitivity:
96.5% (95% Cl: 91.3% - 98.6%)
GAS Specificity:
98.0% (95% Cl: 97.0% - 98.6%)
Pyo GCS/GGS Sensitivity:
95.7% (95% Cl: 88.1% - 98.5%)
Pyo GCS/GGS Specificity:
98.3% (95% Cl: 97.4% - 98.9%)

REPRODUCIBILITY

Three (3) investigative sites assessed the device's inter-site, inter-day and interlintra-assay reproducibility. Each of the laboratories tested a panel of six (6) members that included contrived samples at low positive, (approximately 1 X LoD) and moderately positive sample (approximately 3 X LoD) for M1 and M3 serotypes of Group A Streptococus, a positive and negative control were also included in the panel. The assays were performed in triplicate (3) on five (5) different days. Each site had two (2) operators who each assayed the sample panel once (1) per day, for a total of two (2) sets of data per day. Combined results for all sites are presented in the tables below.

Testing Site 1		Testing Site 2		Testing Site 3			Overall
Sample
Panel
Member	%
Agreement
with
Expected
Results	Avg.
Ct	%
CV	%
Agreement
with
Expected
Results	Avg.
Ct	%
CV	%
Agreement
with
Expected
Results	Avg.
Ct	%
CV	Total %	95% CI
Negative	100.0%
(30/30)	NA	NA	96.7%
(29/30)	41.9	NA	100.0%
(30/30)	NA	NA	98.9%
(89/90)	94.0 to
99.8%
Positive
Control
(PC)	100.0%
(30/30)	29.5	1.4	100.0%
(30/30)	29.5	3	100.0%
(30/30)	30	1.2	100.0%
(90/90)	95.9 to
100.0%
Group A
Strep M1
Serotype
Low
Positive	100.0%
(30/30)	38.6	4.4	83.3%
(25/30)	39.4	5.6	96.7%
(29/30)	38	3.5	93.3%
(84/90)	86.2 to
96.9%
Group A
Strep M1
Serotype
Moderate
Positive	100.0%
(30/30)	35.9	2.6	93.3%
(28/30)	36.3	4.8	100.0%
(30/30)	36	1.8	97.8%
(88/90)	92.3 to
99.4%
Group A
Strep M3
Serotype
Low
Positive	96.7%
(29/30)	38.5	2.9	90.0%
(27/30)	39.2	4.8	96.7%
(29/30)	39	4.5	94.4%
(85/90)	87.6 to
97.6%
Group A
Strep M3
Serotype
Moderate
Positive	100.0%
(30/30)	36.3	2.6	90.0%
(27/30)	36.7	3.2	100.0%
(30/30)	37	2.7	96.7%
(87/90)	90.7 to
98.9%
All	99.4% (179/180)	NA	NA	92.2% (166/180)	NA	NA	98.9% (178/180)	NA	NA	96.9%
(523/540)	95.0 to
98.0%

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Image /page/6/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in bold, black letters, with the word "Diagnostics" in a smaller font size underneath. A curved, black shape is positioned to the left of the word "FOCUS", creating a visual element that draws the eye to the brand name.

510(k) Summary

Simplexa™ Group A Strep Direct Catalog No. MOL2850 Simplexa™ Group A Strep Positive Control Pack Catalog No. MOL2860 March 17, 2015 Page 4 of 10

ANALYTICAL SENSITIVITY/LIMIT OF DETECTION

The Limit of Detection (LoD) was determined for the Simplexa™ Group A Strep Direct by performing a dilution series. The LoD samples used for the study were contrived using two (2) serotypes of Group A Streptococcus; M1 and M3 that were verified (re-grown and re-titered) bacterial stock. For each serotype, eight (8) different concentrations were spiked in simulated matrix from the verified bacterial and confirmed using thirty two (32) replicates. The Limit of Detection (LoD) was determined to be the following;

Simplexa™ Group A Strep Direct - Limit of Detection
Group A Strep Serotype	Concentration (cfu/mL)
M1	682
M3	2350

ANALYTICAL REACTIVITY / CROSS REACTIVITY

Analytical Reactivity

Analytical Reactivity was assessed for the ability of Simplexa™ Group A Strep Direct to detect sixty (60) Group A Streptococus strains not present in the LoD study. Analytical Reactivity was observed in testing of twenty one (21) strains. All were detected as positive for Group A Strep at or below 5000 cfu/mL. Analytical Reactivity was tested using in silico NCBI BLAST sequence analysis methods for thirty nine (39) GAS strains distinct from the wet tested strains.

| Streptococcus pyogenes
Serotype | Concentration (cfu/mL) | Simplexa™ Direct Qualitative Result
% Detection |
|------------------------------------|----------- | M2 | 1.50 X 103 | M4 | 1.50X 103 | M5 | 1.50 X 103 | M6 | 1.50 X 103 | M9 | 3.00 X 103 | M12 | 1.50 X 103 | M13 | 1.50 X 103 | M14 | 1.50 X 103 | M18 | 5.00 X 103 | M22 | 1.50 X 103 | M27 | 3.00 X 103 | M28 | 1.50 X 103 | M29 | 1.50 X 103 | M49 | 1.50 X 103 | M73 | 3.00 X 103 | M75 | 3.00 X 103 | M77 | 3.00 X 103 | M78 | 1.50 X 103 Group A Strep
-------------|---------------------------------------------------------------------|
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |
| 100% (3/3) |

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Image /page/7/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo features the word "FOCUS" in bold, sans-serif font. Below "FOCUS" is the word "Diagnostics" in a smaller, less bold font. A curved, black shape is on the left side of the word "FOCUS", adding a design element to the logo.

510(k) Summary

Simplexa™ Group A Strep Direct Catalog No. MOL2850 Simplexa™ Group A Strep Positive Control Pack Catalog No. MOL2860 March 17, 2015 Page 5 of 10

| Streptococcus pyogenes
Serotype | Concentration (cfu/mL) | Simplexa™ Group A Strep
Direct Qualitative Result
% Detection |
|-------------------------------------------|------------------------|---------------------------------------------------------------------|
| M82 | $1.50 X 10^3$ | 100% (3/3) |
| M87 | $1.50 X 10^3$ | 100% (3/3) |
| M89 | $3.00 X 10^3$ | 100% (3/3) |

Cross Reactivity (Analytical Specificity)

Analytical reactivity was evaluated for the Simplexa™ Group A Strep the ability to exclusively identify Group A Streptococcus with no cross reactivity to organisms that are closely related, or cause similar clinical symptoms, or present as normal flora in the pharynx. Negative specimens were spiked with potentially cross-reactive organisms at known concentrations. Spiked specimens were examined for reactivity with the Group A Strep Direct. Sixty four (64) organisms were tested. Bacteroides ovalis and Tremella fuciformis were not available for testing therefore data from In-Silico NCBI BLAST sequence analysis was performed. No cross reactivity was found

| Cross Reactant | Tested Concentration | Simplexa™ Group A Strep Direct
Qualitative Results: %Detection
(# Detected/#Tested) |
|---------------------------------------------------|---------------------------|-------------------------------------------------------------------------------------------|
| Baseline1 | Not Applicable | 0.0% (0/5) |
| Baseline2 | Not Applicable | 0.0% (0/5) |
| Adenovirus 1 | 1.00 X 105 TCID50/mL | 0.0% (0/3) |
| Adenovirus 7A | 1.00 X 105 TCID50/mL | 0.0% (0/3) |
| Arcanobacterium haemolyticum | 1.00 X 106 TCID50/mL | 0.0% (0/3) |
| Bacillus cereus | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Bacteroides ovatus | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Bordetella pertussis | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Burkholderia cepacia | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Campylobacter rectus | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Candida albicans | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Chlamydia pneumoniae | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Coronavirus 229E | 1.00 X 105 TCID50/mL | 0.0% (0/3) |
| Corynebacterium diphtheriae | 1.00 X 106 TCID50/mL | 0.0% (0/3) |
| Cytomegalovirus (CMV) | 1.00 X 105 TCID50/mL | 0.0% (0/3) |
| Enterococcus faecalis vanB | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Enterovirus 71 | 1.00 X 105 TCID50/mL | 0.0% (0/3) |
| Epstein-Barr virus (B95-8) | 1.00 X 105 copies/mL | 0.0% (0/3) |
| Escherichia coli | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Fusobacterium necrophorum | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Haemophilus influenzae | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Cross Reactant | Tested Concentration | Simplexa™ Group A Strep Direct
Qualitative Results: %Detection
(# Detected/#Tested) |
| HSV-1 McIntyre | 1.00 X 105 TCID50/mL | 0.0% (0/3) |
| HSV-2 G | 1.00 X 105 TCID50/mL | 0.0% (0/3) |
| Influenza A/Hong Kong/8/68 H3N2 | 1.00 X 105 TCID50/mL | 0.0% (0/3) |
| Influenza B/Panama/45/90 | 1.00 X 105 TCID50/mL | 0.0% (0/3) |
| Klebsiella pneumoniae | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Lactobacillus acidophilus | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Legionella pneumophila | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Metapneumovirus-9 | 1.00 X 105 TCID50/mL | 0.0% (0/3) |
| Moraxella catarrhalis | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Mycoplasma pneumoniae | 1.00 X 106 CCU/ml | 0.0% (0/3) |
| Neisseria gonorrhoeae | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Neisseria meningitidis | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Parainfluenza 1 | 1.00 X 105 TCID50/mL | 0.0% (0/3) |
| Parainfluenza 2 | 1.00 X 105 TCID50/mL | 0.0% (0/3) |
| Parainfluenza 3 | 1.00 X 105 TCID50/mL | 0.0% (0/3) |
| Peptostreptococcus micros | 1.00 X 106 cfu/mL | 0.0% (0/6) |
| Pseudomonas aeruginosa | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Rhinovirus 1A | 1.00 X 105 TCID50/mL | 0.0% (0/3) |
| RSV-B 9320 | 1.00 X 105 TCID50/mL | 0.0% (0/3) |
| Staphylococcus aureus (MRSA), ATCC
43300 | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Staphylococcus epidermidis (MRSE), ATCC
29887 | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Stenotrophomonas maltophilia | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus agalactiae | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus anginosus | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus canis | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus constellatus subsp.
constellatus | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus cristatus | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus dysgalactiae | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus equi subsp. zooepidemicus | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus equinus | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus gallolyticus | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus gordonii | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus intermedius | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus mitis | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Cross Reactant | Tested Concentration | Simplexa™ Group A Strep Direct
Qualitative Results: %Detection
(# Detected/#Tested) |
| Streptococcus mutans | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus oralis | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus parasanguinis | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus pneumoniae | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus salivarius | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus sanguinis | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus sobrinus | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus uberis | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Streptococcus vestibularis | 1.00 X 106 cfu/mL | 0.0% (0/3) |
| Treponema denticola | 1.00 X 106 spirochetes/mL | 0.0% (0/3) |
| Veillonella parvula | 1.00 X 106 cfu/mL | 0.0% (0/3) |

8

Image /page/8/Picture/0 description: The image shows the logo for Focus Diagnostics. The word "FOCUS" is in large, bold, sans-serif font. A curved, swooping line extends from the top left of the "F" and arches over the word. Below "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font with a line above it.

510(k) Summary

Simplexa™ Group A Strep Direct Catalog No. MOL2850 Simplexa™ Group A Strep Positive Control Pack Catalog No. MOL2860 March 17, 2015 Page 6 of 10

9

Image /page/9/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in bold, sans-serif font. A curved line extends from the left side of the "F", arching over the word. Below "FOCUS" is the word "Diagnostics" in a smaller, non-bold font, underlined with a thin line.

510(k) Summary

Simplexa™ Group A Strep Direct Catalog No. MOL2850 Simplexa™ Group A Strep Positive Control Pack Catalog No. MOL2860 March 17, 2015 Page 7 of 10

INTERFERENCE

The performance of Simplexa™ Group A Strep Direct was evaluated with potentially interfering substances that may be present in pharynx. The potentially interfering substances were evaluated in a contrived sample that contained Group A Streptococus at low positive concentrations (approximately 1 X LoD) and moderate positive concentrations (approximately 3 X LoD) of Group A Streptococus serotypes M1 and M3. There was no evidence of interference caused by the substances at the concentrations tested.

| Potential Interferent | Active Ingredient | Interferent
Concentration | GAS Bacterial
Serotype | Simplexa™ Group
A Strep Direct
Qualitative Result
(# Detected/#
Tested) |
|----------------------------------------------------|---------------------------------------------------------------------------------------------------------------------|----------------------------------------|---------------------------|-------------------------------------------------------------------------------------|
| Baseline | Not Applicable | Not Applicable | M1 | 100.0%(5/5) |
| Baseline | Not Applicable | Not Applicable | M3 | 100.0%(5/5) |
| Afrin nasal Spray | Oxymetazoline
Hydrochloride | 15% v/v | M1 | 100.0%(3/3) |
| Afrin nasal Spray | Oxymetazoline
Hydrochloride | 15% v/v | M3 | 100.0%(3/3) |
| Antibiotic | Amoxicillin | 0.5 mg/mL | M1 | 100.0%(3/3) |
| Antibiotic | Amoxicillin | 0.5 mg/mL | M3 | 100.0%(3/3) |
| Antibiotic | Cephalexin | 0.04 mg/mL | M1 | 100.0%(3/3) |
| Antibiotic | Cephalexin | 0.04 mg/mL | M3 | 100.0%(3/3) |
| Antibiotic | Clindamycin | 0.06 mg/mL | M1 | 100.0%(5/5) |
| Antibiotic | Clindamycin | 0.06 mg/mL | M3 | 100.0%(3/3) |
| Antibiotic | Erythromycin | 1 mg/mL | M1 | 100.0%(3/3) |
| Antibiotic | Erythromycin | 1 mg/mL | M3 | 100.0%(3/3) |
| Antibiotic | Penicillin | 1200 U/mL | M1 | 100.0%(3/3) |
| Antibiotic | Penicillin | 1200 U/mL | M3 | 100.0%(3/3) |
| Aspirin | Aspirin | 0.62 mg/mL | M1 | 100.0%(3/3) |
| Potential Interferent | Active Ingredient | Interferent
Concentration | GAS Bacterial
Serotype | Simplexa™ Group
A Strep Direct
Qualitative Result
(# Detected/#
Tested) |
| Aspirin | Aspirin | 0.62 mg/mL | M3 | 100.0%(3/3) |
| Benadryl | Diphenhydramine
HCl | 10 µL/swab | M1 | 100.0%(3/3) |
| Benadryl | Diphenhydramine
HCl | 10 µL/swab | M3 | 100.0%(3/3) |
| Blood | Blood | 10% v/v | M1 | 100.0%(3/3) |
| Blood | Blood | 10% v/v | M3 | 100.0%(3/3) |
| Chloraseptic Sore Throat
Spray | Phenol | 10% v/v | M1 | 100.0%(3/3) |
| Chloraseptic Sore Throat
Spray | Phenol | 10% v/v | M3 | 100.0%(3/3) |
| Contac Cold + Flu tablet | Acetaminophen
Chlorpheniramine
Maleate
Phenylephrine HCI | 16.2 mg/mL
0.06 mg/mL
0.16 mg/mL | M1 | 100.0%(3/3) |
| Contac Cold + Flu tablet | Acetaminophen
Chlorpheniramine
Maleate
Phenylephrine HCI | 16.2 mg/mL
0.06 mg/mL
0.16 mg/mL | M3 | 100.0%(3/3) |
| Corticosterone | Corticosterone | 4 mg/swab | M1 | 100.0%(3/3) |
| Corticosterone | Corticosterone | 4 mg/swab | M3 | 100.0%(3/3) |
| Crest Complete
Toothpaste | Sodium Fluoride | 0.1 mg/mL | M1 | 100.0%(3/3) |
| Crest Complete
Toothpaste | Sodium Fluoride | 0.1 mg/mL | M3 | 100.0%(3/3) |
| Finafta Oral Anesthetic /
Analgesics | Ethyl Alcohol
Salicylic Acid
Benzocaine | 1/10X dilution | M1 | 100.0%(3/3) |
| Finafta Oral Anesthetic /
Analgesics | Ethyl Alcohol
Salicylic Acid
Benzocaine | 1/10X dilution | M3 | 100.0%(3/3) |
| Listerine | Eucalyptol
Menthol
Methyl Salicylate
Thymol | 10 µL/swab | M1 | 100.0%(3/3) |
| Listerine | Eucalyptol
Menthol
Methyl Salicylate
Thymol | 10 µL/swab | M3 | 100.0%(3/3) |
| Mucin | Purified Mucin
Protein | 60 µg/mL | M1 | 100.0%(3/3) |
| Mucin | Purified Mucin
Protein | 60 µg/mL | M3 | 100.0%(3/3) |
| Neo-Synephrine | Phenylephrine HCI | 15% v/v | M1 | 100.0%(3/3) |
| Neo-Synephrine | Phenylephrine HCI | 15% v/v | M3 | 100.0%(3/3) |
| Potential Interferent | Active Ingredient | Interferent
Concentration | GAS Bacterial
Serotype | Simplexa™ Group
A Strep Direct
Qualitative Result
(# Detected/#
Tested) |
| Nyquil | Dextromethorphan
Hydrobromide
Doxylamine
Succinate | 1/200 X dilution | M1 | 100.0%(3/3) |
| Nyquil | Dextromethorphan
Hydrobromide
Doxylamine
Succinate | 1/200 X dilution | M3 | 100.0%(3/3) |
| Pain Medication | NSAID | 0.1 mg/mL | M1 | 100.0%(3/3) |
| Pain Medication | NSAID | 0.1 mg/mL | M3 | 100.0%(3/3) |
| Pain Medication | Tylenol | 1 mg/mL | M1 | 100.0%(3/3) |
| Pain Medication | Tylenol | 1 mg/mL | M3 | 100.0%(3/3) |
| Robitussin Cough / Chest
Congestion Cough Syrup | Dextromethorphan
HBr
Guaifenesin | 2.0 mg/mL | M1 | 100.0%(3/3) |
| Robitussin Cough / Chest
Congestion Cough Syrup | Dextromethorphan
HBr
Guaifenesin | 2.0 mg/mL | M3 | 100.0%(3/3) |
| Saline Nasal Spray | Sodium Chloride
with Preservatives | 15% v/v | M1 | 100.0%(3/3) |
| Saline Nasal Spray | Sodium Chloride
with Preservatives | 15% v/v | M3 | 100.0%(3/3) |
| Saliva | Water, Electrolytes,
Mucus, Etc. | 50 µL/swab | M1 | 100.0%(3/3) |
| Saliva | Water, Electrolytes,
Mucus, Etc. | 50 µL/swab | M3 | 100.0%(3/3) |
| Scope | Glycerin
Sodium Saccharin
Sodium Benzoate
Cetylpyridinum
Chloride
Benzoic Acid
Blue 1
Yellow 5 | 10 µL/swab | M1 | 100.0%(3/3) |
| Scope | Glycerin
Sodium Saccharin
Sodium Benzoate
Cetylpyridinum
Chloride
Benzoic Acid
Blue 1
Yellow 5 | 10 µL/swab | M3 | 100.0%(3/3) |
| Sore Throat Lozenge | Menthol | 1.7 mg/mL | M1 | 100.0%(3/3) |
| Sore Throat Lozenge | Menthol | 1.7 mg/mL | M3 | 100.0%(3/3) |
| Sore Throat Lozenge | Pectin | 0.34 mg/mL | M1 | 100.0%(3/3) |
| Sore Throat Lozenge | Pectin | 0.34 mg/mL | M3 | 100.0%(3/3) |
| Potential Interferent | Active Ingredient | Interferent
Concentration | GAS Bacterial
Serotype | Simplexa™ Group
A Strep Direct
Qualitative Result
(# Detected/#
Tested) |
| Sore Throat Lozenge | Zinc Gluconate Glycine | 0.1 mg/mL | M1 | 100.0%(3/3) |
| Sore Throat Lozenge | Zinc Gluconate Glycine | 0.1 mg/mL | M3 | 100.0%(3/3) |
| Thymol | Thymol | 400 mg/swab | M1 | 100.0%(3/3) |
| Thymol | Thymol | 400 mg/swab | M3 | 100.0%(3/3) |
| Zicam Oral Mist | Zincum Aceticum Zincum Gluonicum | 0.625% v/v | M1 | 100.0%(3/3) |
| Zicam Oral Mist | Zincum Aceticum Zincum Gluonicum | 0.625% v/v | M3 | 100.0%(3/3) |

10

Image /page/10/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo features the word "FOCUS" in bold, sans-serif font. A curved line extends from the left side of the "F", arching over the word. Below "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin line.

510(k) Summary

Simplexa™ Group A Strep Direct Catalog No. MOL2850 Simplexa™ Group A Strep Positive Control Pack Catalog No. MOL2860 March 17, 2015 Page 8 of 10

11

Image /page/11/Picture/0 description: The image shows the logo for Focus Diagnostics. The word "FOCUS" is in large, bold, sans-serif font. There is a curved line above and to the left of the word. Below the word "FOCUS" is a horizontal line, and below that is the word "Diagnostics" in a smaller font.

510(k) Summary

Simplexa™ Group A Strep Direct Catalog No. MOL2850 Simplexa™ Group A Strep Positive Control Pack Catalog No. MOL2860 March 17, 2015 Page 9 of 10

12

Image /page/12/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in bold, sans-serif font, with the word "Diagnostics" in a smaller font size underneath. A curved, swooping line extends from the left side of the image, arching over the word "FOCUS", creating a visual element that emphasizes the brand name.

510(k) Summary

Simplexa™ Group A Strep Direct Catalog No. MOL2850 Simplexa™ Group A Strep Positive Control Pack Catalog No. MOL2860 March 17, 2015 Page 10 of 10

CLINICAL PROSPECTIVE STUDY

One thousand three hundred and ninety seven (1397) samples that were prospectively collected from four (4) geographically diverse sites between May 6, 2014 from patients with signs and symptoms of Group A Strep infections of the pharynx. One thousand three hundred and fifty two (1352) samples were evaluable on Simplexa™ Group A Strep Direct and the comparator culture method. Samples were tested on Simplexa™ Group A Strep Direct at the collection sites and comparator culture method was performed at one (1) central laboratory. The invalid rate of the clinical prospective study using Simplexa™ Group A Strep Direct was 0.57% (eight out of one thousand three hundred and ninety six 8/1396 samples). Discrepant analysis was performed using a validated bidirectional sequencing assay.

Clinical Prospective Study: (Overall)
Simplexa™
Group A Strep
Direct Result	Culture Method
Detected	Not Detected	Total
Detected	152	57a	209
Not Detected	4b	1139	1143
Total	156	1196	1352
%Sensitivity	97.4%(152/156)
95% CI: 93.6% to
99.0%	%Specificity	95.2%(1139/1196)
95% CI: 93.9% to
96.3%
% Positive
Predictive Value
(PPV)	72.7%(152/209)
95% CI: 66.3% to
78.3%	%Negative Predictive
Value (NPV)	99.7%(1139/1143)
95% CI: 99.1% to
99.9%
a46/57 discrepant samples were Group A Strep Positive, 9/57 were "Group A Strep
Negative and 2/57 were Indeterminate when tested using a validated bidirectional
sequencing assay.
b2/ 4 discrepant samples were Group A Strep Positive and 2/4 were Group A Strep
Negative when tested using a validated bidirectional sequencing assay.

CONCLUSION

The conclusions drawn from the nonclinical tests demonstrate the device is as safe and effective as the legally marketed device identified above.