Who is the manufacturer of Simplexa Flu A/B & RSV Direct & Simplexa Flu A/B RSV Positive Control Pack?

Focus Diagnostics filed the 510(k) submission for Simplexa Flu A/B & RSV Direct & Simplexa Flu A/B RSV Positive Control Pack (K142365).

What is the FDA product code for Simplexa Flu A/B & RSV Direct & Simplexa Flu A/B RSV Positive Control Pack?

OCC. It is classified under 21 CFR 866.3980 as a Class 2 device in the Microbiology panel.

What is the regulatory pathway for Simplexa Flu A/B & RSV Direct & Simplexa Flu A/B RSV Positive Control Pack?

510(k) clearance (submission type: Traditional).

Who can help prepare an FDA 510(k) submission like Simplexa Flu A/B & RSV Direct & Simplexa Flu A/B RSV Positive Control Pack?

Innolitics — a medical-device software consultancy — provides regulatory and engineering support for FDA 510(k) submissions. See https://innolitics.com/services/510ks/ or contact https://innolitics.com/contact.

Simplexa Flu A/B & RSV Direct & Simplexa Flu A/B RSV Positive Control Pack

K142365 · Focus Diagnostics · OCC · Dec 5, 2014 · Microbiology

Device Facts

Record ID	K142365
Device Name	Simplexa Flu A/B & RSV Direct & Simplexa Flu A/B RSV Positive Control Pack
Applicant	Focus Diagnostics
Product Code	OCC · Microbiology
Decision Date	Dec 5, 2014
Decision	SESE
Submission Type	Traditional
Regulation	21 CFR 866.3980
Device Class	Class 2

Intended Use

The Focus Diagnostics Simplexa™ Flu A/B & RSV Direct assay is intended for use on the 3M Integrated Cycler instrument for the in vitro qualitative detection and differentiation of influenza A virus, influenza B virus, and respiratory syncytial virus (RSV) RNA in nasopharyngeal swabs (NPS) from human patients with signs and symptoms of respiratory tract infection in conjunction with clinical and epidemiological risk factors. This test is intended for use as an aid in the differential diagnosis of influenza A, influenza B, and RSV viral infections in humans and is not intended to detect influenza C. Negative results do not preclude influenza virus or RSV infection and should not be used as the sole basis for treatment or other patient management decisions. Performance characteristics for influenza A were established with clinical specimens collected during the 2010/2011 influenza season when 2009 H1N1 influenza and H3N2 were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging, performance characteristics may vary. If infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent Influenza viruses and sent to the state or local health department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens. Focus Diagnostics' Simplexa™ Flu A/B & RSV Positive Control Pack is intended to be used as a control with the Simplexa™ Flu A/B & RSV Direct kit. This control is not intended for use with other assays or systems.

Device Story

Real-time RT-PCR system for direct amplification, detection, and differentiation of influenza A, influenza B, and RSV RNA from unprocessed nasopharyngeal swabs; eliminates nucleic acid extraction. System components: Simplexa™ Flu A/B & RSV Direct assay, 3M Integrated Cycler, Integrated Cycler Studio Software, and Direct Amplification Disc. Bi-functional fluorescent probe-primers and reverse primers target conserved matrix genes (Flu A, Flu B) and M gene (RSV); RNA internal control monitors RT-PCR failure/inhibition. Used in clinical settings by trained personnel. Output: qualitative detection of viral RNA. Results aid clinicians in differential diagnosis of respiratory infections. Benefits: rapid, direct testing without extraction.

Clinical Evidence

Bench testing only. Method comparison study using 265 archived clinical samples (55 Flu A+, 55 Flu B+, 55 RSV+, 100 negative) compared Gen 2.0/2.1 to Gen 1.0. Results showed high PPA (97.8-100%) and NPA (95.7-100%) across all targets. Analytical reactivity confirmed detection of diverse influenza strains (H1, H3, H5, H7N9). Analytical specificity (cross-reactivity) tested against 32 common respiratory organisms with no cross-reactivity observed. Interference studies showed no impact from common nasal substances.

Technological Characteristics

Real-time RT-PCR assay. Targets: Flu A (matrix gene, FAM), Flu B (matrix gene, JOE), RSV (M gene, CFR610), Internal Control (N gene, Q670). Form factor: Direct Amplification Disc for 3M Integrated Cycler. Software: Integrated Cycler Studio Software v4.2+. Reagents: DNA polymerase, Reverse Transcriptase, RNase inhibitor, dNTPs, fluorescent probe-primers. Storage: -10 to -30°C.

Indications for Use

Indicated for qualitative detection and differentiation of influenza A, influenza B, and RSV RNA in nasopharyngeal swabs from symptomatic patients as an aid in differential diagnosis. Not for influenza C detection.

Regulatory Classification

Identification

A respiratory viral panel multiplex nucleic acid assay is a qualitative in vitro diagnostic device intended to simultaneously detect and identify multiple viral nucleic acids extracted from human respiratory specimens or viral culture. The detection and identification of a specific viral nucleic acid from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory viral infection when used in conjunction with other clinical and laboratory findings. The device is intended for detection and identification of a combination of the following viruses:(1) Influenza A and Influenza B; (2) Influenza A subtype H1 and Influenza A subtype H3; (3) Respiratory Syncytial Virus subtype A and Respiratory Syncytial Virus subtype B; (4) Parainfluenza 1, Parainfluenza 2, and Parainfluenza 3 virus; (5) Human Metapneumovirus; (6) Rhinovirus; and (7) Adenovirus.

Special Controls

*Classification.* Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay;” (2) For a device that detects and identifies Human Metapneumovirus, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Human Metapneumovirus (hMPV) Using Nucleic Acid Assays;” and (3) For a device that detects and differentiates Influenza A subtype H1 and subtype H3, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Detection and Differentiation of Influenza A Virus Subtypes Using Multiplex Nucleic Acid Assays.” See § 866.1(e) for the availability of these guidance documents.

Predicate Devices

Simplexa™ Flu A/B & RSV Direct and Simplexa™ Flu A/B & RSV Positive Control Pack (K120413)

Related Devices

K152408 — Simplexa Flu A/B & RSV Direct and Simplexa Flu A/B & RSV Positive Control Pack · Focus Diagnostics, Inc. · Sep 21, 2015
K120413 — SIMPLEXA FLU A/B & RSV DIRECT, SIMPLEXA FLU A/B & RSV POSITIVE CONTROL PACK · Focus Diagnostics, Inc. · Jul 13, 2012
K102170 — SIMPLEXA FLU A/B & RSV · Focus Diagnostics, Inc. · Nov 24, 2010
K213822 — cobas Influenza A/B & RSV nucleic acid test for use on the cobas Liat System · Roche Molecular Systems, Inc. · Jul 6, 2022
K142045 — XPERT FLU/RSV XC ASSAY · Cepheid · Nov 22, 2014

Submission Summary (Full Text)

{0}------------------------------------------------ Image /page/0/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo features a stylized depiction of a human figure in profile, with three overlapping faces suggesting a sense of community and support. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" is arranged in a circular fashion around the figure, emphasizing the organization's name and national affiliation. The logo is simple, clean, and uses a monochromatic color scheme. 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 Street Cypress, CA 90630 December 5, 2014 Re: K142365 Trade/Device Name: Simplexa™ Flu A/B & RSV Direct Simplexa™ Flu A/B & RSV Positive Control Pack Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory Viral Panel Multiplex Nucleic Acid Assay Regulatory Class: Class II Product Code: OCC, OOI Dated: September 26, 2014 Received: September 29, 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 Part 801); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set {1}------------------------------------------------ forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050. If you desire specific advice for your device on our labeling regulation (21 CFR Part 801), 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" (21CFR 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) K142365 #### Device Name Simplexa™ Flu A/B & RSV Direct and Simplexa™ Flu A/B & RSV Positive Control Pack Indications for Use (Describe) Simplexa™ Flu A/B & RSV Direct REF MOL2650 The Focus Diagnostics Simplexa™ Flu A/B & RSV Direct assay is intended for use on the 3M Integrated Cycler instrument for the in vitro qualitative detection and differentiation of influenza B virus, and respiratory syncytial virus (RSV) RNA in nasopharyngeal swabs (NPS) from human patients with signs and symptoms of respiratory tract infection in conjunction with clinical and epidemiological risk factors. This test is intended for use as an aid in the differential diagnosis of influenza B, and RSV viral infections in humans and is not intended to detect influenza C. Negative results do not preclude influenza virus or RSV infection and should not be used as the sole basis for treatment or other patient management decisions. Performance characteristics for influenza A were established with clinical specimens collected during the 2010/2011 influenza season when 2009 H1N1 influenza and H3N2 were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging, performance characteristics may vary. If infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent Influenza viruses and sent to the state or local health department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens. Simplexa™ Flu A/B & RSV Positive Control Pack REF MOL2660 Focus Diagnostics' Simplexa™ Flu A/B & RSV Positive Control Pack is intended to be used as a control with the SimplexaTM Flu A/B & RSV Direct kit. This control is not intended for use with other assays or systems. | Type of Use (Select one or both, as applicable) | | |----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | <div style="display:flex; align-items:center;"> <svg class="bi bi-check-square-fill" fill="currentColor" height="1em" viewbox="0 0 16 16" width="1em" xmlns="http://www.w3.org/2000/svg"> <path d="M2 0a2 2 0 0 0-2 2v12a2 2 0 0 0 2 2h12a2 2 0 0 0 2-2V2a2 2 0 0 0-2-2H2zm10.03 4.97a.75.75 0 0 0-1.08.022L7.477 9.417 5.384 7.323a.75.75 0 0 0-1.06 1.06L6.97 11.03a.75.75 0 0 0 1.079-.02l3.992-4.99a.75.75 0 0 0-.01-1.05z" fill-rule="evenodd"></path> </svg> Prescription Use (Part 21 CFR 801 Subpart D) </div> | <div style="display:flex; align-items:center;"> <svg class="bi bi-square" fill="currentColor" height="1em" viewbox="0 0 16 16" width="1em" xmlns="http://www.w3.org/2000/svg"> <path d="M14 1H2a1 1 0 0 0-1 1v12a1 1 0 0 0 1 1h12a1 1 0 0 0 1-1V2a1 1 0 0 0-1-1zM2 0a2 2 0 0 0-2 2v12a2 2 0 0 0 2 2h12a2 2 0 0 0 2-2V2a2 2 0 0 0-2-2H2z" fill-rule="evenodd"></path> </svg> Over-The-Counter Use (21 CFR 801 Subpart C) </div> | CONTINUE ON A SEPARATE PAGE IF NEEDED. This section applies only to requirements of the Paperwork Reduction Act of 1995. ***DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.*** The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to: Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff(@fda.hhs.gov "An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number." {3}------------------------------------------------ Image /page/3/Picture/0 description: The image shows the logo for Focus Diagnostics. The word "FOCUS" is in large, bold, black letters. To the left of the word is a curved black line that starts thin and gets thicker as it curves around the word. Below the word "FOCUS" is the word "Diagnostics" in a smaller, less bold font, with a horizontal line above it. 510(k) Summarv Simplexa" Flu A/B & RSV Direct REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 1 of 14 | 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 | December 2, 2014 | | Proprietary Name | Simplexa™ Flu A/B & RSV Direct and Simplexa™ Flu A/B & RSV Positive Control Pack | | Generic Name | Respiratory viral panel nucleic acid | | Classification | Class II | | US Product Code | OCC - Respiratory Viral Panel Nucleic Acid Assay System OOI - Real Time Nucleic Acid Amplification System | | Regulation Number | 21 CFR § 866.3980 | | Predicate Device | K120413 Simplexa™ Flu A/B & RSV Direct and Simplexa™ Flu A/B & RSV Positive Control Pack | #### 510(k) SUBMISSION PURPOSE The purpose of the submission is to add eight (8) additional influenza strains to the Analytical Reactivity of the Simplexa™ Flu A/B & RSV Direct. The package insert was updated to include the additional strains which included the following six (6) additional strains of Influenza A and two (2) additional strains of Influenza A/Anhui/1/2013, Influenza A/California/12/2012 (H1N1) pdm09, Influenza A/Indiana/08/2011 (H3N2)v, Influenza A/Minnesota/11/2010 (H3N2)v, Influenza A/Ohio/02/2012 (H3N2), Influenza A/Texas/50/2012 (H3N2), Influenza B/Brisbane/60/2008 and Influenza B/Wisconsin/01/2010. Changes to the Simplexa™ Flu A/B and RSV Direct K120413 (Gen 1) were made in two phases. The first phase of the modifications (Gen 2.0) included changes to the reaction mix formulation and cycling conditions: (a) change in the annealing temperature (decreased), (b) change in the (increased), (c) change in the enzyme used, and (d) change to the RSV cut-off Ct (decreased). Limited changes (Gen. 2.1) were also made to the manufacturing process and materials to increase the stability of the reaction mix and to minimize non-specific products. #### INTENDED USE #### Simplexa™ Flu A/B & RSV Direct REF MOL2650 The Focus Diagnostics Simplexa™ Flu A/B & RSV Direct assay is intended for use on the 3M Integrated Cycler instrument for the in vitro qualitative detection and differentiation of influenza B virus, and respiratory syncytial virus (RSV) RNA in nasopharyngeal swabs (NPS) from human patients and symptoms of respiratory tract infection in conjunction with clinical and epidemiological risk factors. This test is intended for use as an aid in the differential diagnosis of influenza B, and RSV viral infections in humans and is not intended to detect influenza C. Negative results do not preclude influenza virus or RSV infection and should not be used as the sole basis for treatment or other patient management decisions. Performance characteristics for influenza A were established with clinical specimens collected during the 2010/2011 influenza season when 2009 H1N1 influenza and H3N2 were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging, performance characteristics may vary. {4}------------------------------------------------ Image /page/4/Picture/0 description: The image shows the logo for Focus Diagnostics. The word "FOCUS" is in large, bold, sans-serif font. To the left of the word "FOCUS" is a curved, black shape that resembles a stylized check mark. Below the word "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin black line. ### 510(k) Summary Simplexa" Flu A/B & RSV Direct REF MOL2650 Simplexa" Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 2 of 14 If infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent Influenza viruses and sent to the state or local health department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens. #### Simplexa™ Flu A/B & RSV Positive Control Pack REF MOL2660 Focus Diagnostics' Simplexa™ Flu A/B & RSV Positive Control Pack is intended to be used as a control with the Simplexa™ Flu A/B & RSV Direct kit. This control is not intended for use with other assays or systems. #### DEVICE DESCRIPTION The Simplexa™ Flu A/B & RSV Direct assay system is a real-time RT-PCR system that enables the direct amplification, detection and differentiation of human influenza A (Flu A) virus RNA, human influenza B (Flu B) virus RNA and RSV RNA from unprocessed nasopharyngeal swabs that have not undergone nucleic acid extraction. The system consists of the Simplexa™ Flu A/B & RSV Direct assay, the 3M Integrated Cycler (with Integrated Cycler Studio Software), the Direct Amplification Disc and associated accessories. In the Simplexa™ Flu A/B & RSV Direct assay, bi-functional fluorescent probe-primers are used together with corresponding reverse primers to amplify Flu A, Flu B, RSV and internal control RNA. The assay provides three results; conserved regions of influenza A viruses (matrix gene), influenza B viruses (matrix gene) and RSV (M gene) are targeted to identify these viruses in the specimen. An RNA internal control is used to detect RT-PCR failure and/or inhibition. #### MATERIALS PROVIDED The Focus Diagnostics Simplexa™ Flu A/B & RSV Direct kit contains sufficient reagents for 24 reactions. Upon receipt, store at -10 to -30 ℃ (do not use a frost-free freezer). Each vial contains sufficient material for one use. Use within 30 minutes of removing from the freezer. | Component Name | REF | EC SYMBOL ON LABEL | Abbreviated Name | Cap Color | Number of Vials | Reactions per Vial/Kit | Volume per Vial | |------------------------------------------------|---------|-----------------------|---------------------|--------------|--------------------|---------------------------|--------------------| | Simplexa™ Flu A/B & RSV Direct Reaction Mix | MOL2651 | REAG A | RM | Brown | 24 | 1/24 | 50 μL | #### Component Description | Kit Component | Contents | | | | | | | | | | | | | | | | | | | | | | | | | | |--------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--|--| | | DNA polymerase, Reverse Transcriptase, RNase inhibitor, buffer and dNTPs, encapsulated RNA Template Dye-labeled fluorescent primers specific for detection of influenza A, influenza B and RSV and for the Internal Control | | | | | | | | | | | | | | | | | | | | | | | | | | | | Target Probe Fluorophore (Dye) Excitation Emission Targeted Gene Flu A FAM 495 520 matrix Flu B JOE 520 548 matrix RSV CFR610 590 610 M gene Internal Control "RNA IC" Q670 644 670 N/A | | | | | | | | | | | | | | | | | | | | | | | | | | | Simplexa™ Flu A/B & RSV Direct Reaction Mix (RM) | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | | Simplexa™ Flu A/B & RSV Direct Barcode Card | Assay specific parameters, lot number, expiration date | | | | | | | | | | | | | | | | | | | | | | | | | | Kit Description {5}------------------------------------------------ Image /page/5/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. To the left of the word "FOCUS" is a curved, black shape that resembles a check mark or a swoosh. ### 42365 510(k) Summary Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa"" Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 3 of 14 ### Simplexa™ Flu A/B & RSV Positive Control Pack REF MOL2660 Product Description | Component Name | REF | Description | Cap Color | Number of Vials | Reactions per Vial/Kit | Volume per Vial | |----------------------------------------------------|---------|-------------------------------------------------------------------------------------|-----------|-----------------|------------------------|-----------------| | Simplexa™ Flu A/B & RSV Direct Positive Control | MOL2661 | Inactivated influenza A virus, Inactivated influenza B virus, Inactivated RSV | Red | 10 | 1/10 | 100 µL | ### MATERIALS SUPPLIED SEPARATELY 1. Direct Amplification Disc Kit (REF MOL1455) - Direct Amplification Discs for use on the Integrated Cycler. a. #### INSTRUMENT REQUIREMENTS 3M Integrated Cycler with Integrated Cycler Studio Software version 4.2 or higher #### COMPARISON TO PREDICATE ### Similarities | Feature | Predicate K120413 | Modified Device K142365 | |------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------| | Intended Use | Simplexa™ Flu A/B & RSV Direct REF MOL2650 The Focus Diagnostics Simplexa™ Flu A/B & RSV Direct assay is intended for use on the 3M Integrated Cycler instrument for the in vitro qualitative detection and differentiation of influenza A virus, influenza B virus, and respiratory syncytial virus (RSV) RNA in nasopharyngeal swabs (NPS) from human patients with signs and symptoms of respiratory tract infection in conjunction with clinical and epidemiological risk factors. This test is intended for use as an aid in the differential diagnosis of influenza A, influenza B, and RSV viral infections in humans and is not intended to detect influenza C. Negative results do not preclude influenza virus or RSV infection and should not be used as the sole basis for treatment or other patient management decisions. Performance characteristics for influenza A were established with clinical specimens collected during the 2010/2011 influenza season when 2009 H1N1 influenza and H3N2 were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging, performance characteristics may vary. If infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent | Same | | Feature | Predicate K120413 | Modified Device K142365 | | | Influenza viruses and sent to the state or local health department for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens. | | | | Simplexa™ Flu A/B & RSV Positive Control Pack REF MOL2660 | | | | Focus Diagnostics' Simplexa™ Flu A/B & RSV Positive Control Pack is intended to be used as a control with the Simplexa™ Flu A/B & RSV Direct kit. This control is not intended for use with other assays or systems. | | | Technology | The Simplexa™ Flu A/B & RSV Direct assay system is a real-time RT-PCR system that enables the direct amplification, detection and differentiation of human influenza A (Flu A) virus RNA, human influenza B (Flu B) virus RNA and RSV RNA from unprocessed nasopharyngeal swabs that have not undergone nucleic acid extraction. The system consists of the Simplexa™ Flu A/B & RSV Direct assay, the 3M Integrated Cycler (with Integrated Cycler Studio Software), the Direct Amplification Disc and associated accessories. In the Simplexa™ Flu A/B & RSV Direct assay, bi-functional fluorescent probe-primers are used together with corresponding reverse primers to amplify Flu A, Flu B, RSV and internal control RNA. The assay provides three results; conserved regions of influenza A viruses (matrix gene), influenza B viruses (matrix gene) and RSV (M gene) are targeted to identify these viruses in the specimen. An RNA internal control is used to detect RT-PCR failure and/or inhibition. | Same | | Instrument | 3M Integrated Cycler | Same | | Specimen Type | Unprocessed nasopharyngeal swabs that have not undergone nucleic acid extraction | Same | | Influenza A Viral Target | Well conserved region of the matrix gene | Same | | Influenza B Viral Target | Well conserved region of the matrix gene | Same | | Respiratory Syncytial Viral Target | M gene | Same | | Assay Type | Qualitative | Same | {6}------------------------------------------------ Image /page/6/Picture/0 description: The image shows the logo for Focus Diagnostics. The word "FOCUS" is in large, bold, sans-serif font. Above and to the left of the word "FOCUS" is a curved, swooping line. Below the word "FOCUS" is the word "Diagnostics" in a smaller font, underlined. ## K142365 510(k) Summary Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa"" Flu A/B & RSV Positive Control Pack [REE] MOL2660 Prepared Date: December 2, 2014 Page 4 of 14 {7}------------------------------------------------ Image /page/7/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 a smaller font, underlined with a thin black line. # K142365 510(k) Summary Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack [REE]MOL2660 Prepared Date: December 2, 2014 Page 5 of 14 Differences {8}------------------------------------------------ Image /page/8/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in a bold, sans-serif font, with the word "Diagnostics" in a smaller font underneath. There is a curved, swooping shape to the left of the word "FOCUS", which adds a dynamic element to the logo. The logo is in black and white. 510(k) Summarv Simplexa" Flu A/B & RSV Direct REF MOL2650 Simplexa" Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 6 of 14 | | Predicate K120413 | Modified Device K142365 | |---------------------------------------------------|-------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | Manufacturing, Material and Cycling Parameters | | (Gen 2.0) - Changes to the annealing temperature (decreased), change in the run time (increased) and changes in enzyme used and change to the RSV cut-off Ct (decreased). | | | | (Gen 2.1) - Changes to manufacturing process and materials. | #### Method Comparison - Simplexa™ Flu AB & RSV Direct Gen 1.0 (K120413) to Simplexa™ Flu AJB & RSV Direct Gen 2.0 The changes implemented for Simplexa™ Flu A/B & RSV Direct (Gen 2.0) were evaluated with archived dinical samples. Based on historical data, 265 samples in Universal Transport Medium (UTM) or Viral Transport Medium (VTM) were assembled with 55 samples positive for influenza A, 55 samples positive for influenza B, 55 samples positive for RSV and 100 samples negative for all of the viruses tested. The 265 samples tested included 131 archived clinical samples originally tested in the clinical study conducted in support of the 510(k) submission (K120413). The other 134 samples included 33 archived samples from the 2010-2011 flu season and 101 samples from the 2013-2014 flu season. The samples were tested in parallel using the Simplexa™ Flu A/B & RSV Direct (Gen 1.0) and the Simplexa™ Flu A/B & RSV Direct (Gen 2.0). The positive percent agreement (PPA) and the negative percent agreement (NPA) between the results obtained with the Simplexa™ Flu A/B & RSV Direct (Gen 1.0) and the Simplexa™ Flu A/B & RSV Direct (Gen 2.0) for each virus are shown below: #### Method Comparison - Simplexa™ Flu A/B & RSV Direct Gen 2.0 to Simplexa™ Flu A/B & RSV Direct Gen 2.1(K142365) The changes implemented for Simplexa™ Flu A/B & RSV Direct (Gen 2.1) were evaluated with archived clinical samples. Based on historical data, 265 samples in Universal Transport Medium (UTM) or Viral Transport Medium (VTM) were assembled with 55 samples positive for influenza A, 55 samples positive for influenza B, 55 samples positive for RSV and 100 samples negative for all of the viruses tested. The 265 samples tested included 125 archived clinical samples originally tested in the clinical study conducted in support of the 510(k) submission (K120413). The other 140 samples included 48 archived samples from the 2010-2011 flu season. 9 samples from 2012-2013 flu season and 83 samples from the 2013-2014 flu season. The samples were tested in parallel using the Simplexa™ Flu A/B & RSV Direct (Gen 2.0) and the Simplexa™ Flu A/B & RSV Direct (Gen 2.1). The positive percent agreement (PPA) and the negative percent agreement (NPA) between the results obtained with the Simplexa™ Flu A/B & RSV Direct (Gen 2.0) and the Simplexa™ Flu A/B & RSV Direct (Gen 2.1) for each virus are shown below: | Simplexa™ Flu A/B & RSV Direct (Gen 2.0) Flu A Result | Simplexa™ Flu A/B & RSV Direct (Gen 1.0) Flu A Result | | Grand Total | | %Agreement | 95% CI | |----------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------|-----------------|----------------|-----|-----------------|-----------------| | | Detected | Not Detected | | | | | | Detected | 58 | 9a | 67 | PPA | 100.0% (58/58) | 93.0% to 100.0% | | Not Detected | 0 | 198 | 198 | NPA | 95.7% (198/207) | 91.9% to 97.7% | | Grand Total | 58 | 207 | 265 | | | | | a7/9 discrepant (K120413 – Negative and K142365 – Positive) samples were positive for Flu A on another FDA cleared NAT. | | | | | | | Influenza A - Simplexa™ Flu A/B & RSV Direct Gen 1.0 (K120413) to Simplexa™ Flu A/B & RSV Direct Gen 2.0 PPA: Positive Percent Agreement, NPA: Negative Percent Agreement {9}------------------------------------------------ Image /page/9/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in a bold, sans-serif font, with a curved, swooping shape extending from the top left of the "F" and arching over the word. Below "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin line. The logo is in black and white. # 510(k) Summary Simplexa™ Flu A/B & RSV Direct REF M Simplexa" Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 7 of 14 Influenza A - Simplexa™ Flu A/B & RSV Direct Gen 2.0 to Simplexa™ Flu A/B & RSV Direct Gen 2.1 (K142365) | Simplexa™ Flu A/B & RSV Direct (Gen 2.1) Flu A Result | Simplexa™ Flu A/B & RSV Direct (Gen 2.0) Flu A Result | | Grand Total | | %Agreement | 95% CI | |----------------------------------------------------------------------|-------------------------------------------------------------|-----------------|----------------|-----|-----------------|-----------------| | | Detected | Not Detected | | | | | | Detected | 58 | 2 | 60 | PPA | 100.0% (58/58) | 93.8% to 100.0% | | Not Detected | 0 | 205 | 205 | NPA | 99.0% (205/207) | 96.5% to 99.7% | | Grand Total | 58 | 207 | 265 | | | | PPA: Positive Percent Agreement, NPA: Negative Percent Agreement Influenza B - Simplexa™Flu A/B & RSV Direct Gen 1.0 (K120413) to Simplexa™ Flu A/B & RSV Direct Gen 2.0 | Simplexa™ Flu A/B & RSV Direct (Gen 2.0) Flu B Result | Simplexa™ Flu A/B & RSV Direct (Gen 1.0) Flu B Result | | Grand Total | | %Agreement | 95% CI | |----------------------------------------------------------------------|-------------------------------------------------------|-----------------|----------------|-----|-----------------|----------------| | | Detected | Not Detected | | | | | | Detected | 54 | 9b | 63 | PPA | 98.2% (54/55) | 90.4% to 99.7% | | Not Detected | 1c | 201 | 202 | NPA | 95.7% (201/210) | 92.1% to 97.7% | | Grand Total | 55 | 210 | 265 | | | | 95/9 discrepant (K120413 – Negative and K142365 – Positive) samples were positive for Flu B on another FDA cleared NAT. °1/1 discrepant (K120413 – Positive and K142365 – Negative) samples was negative for Flu B on another FDA cleared NAT. PPA: Positive Percent Agreement, NPA: Negative Percent Agreement Influenza B - Simplexa™ Flu A/B & RSV Direct Gen 2.0 to Simplexa™ Flu A/B & RSV Direct Gen 2.1 (K142365) | Simplexa™ Flu A/B & RSV Direct (Gen 2.1) Flu B Result | Simplexa™ Flu A/B & RSV Direct (Gen 2.0) Flu B Result | | Grand Total | | %Agreement | 95% CI | |----------------------------------------------------------------------|-------------------------------------------------------------|-----------------|----------------|-----|---------------------|-----------------| | | Detected | Not Detected | | | | | | Detected | 56 | 0 | 63 | PPA | 100.0% (56/56) | 93.6% to 100.0% | | Not Detected | 0 | 209 | 202 | NPA | 100.0% (209/209) | 98.2% to 100.0% | | Grand Total | 56 | 209 | 265 | | | | PPA: Positive Percent Agreement, NPA: Negative Percent Agreement {10}------------------------------------------------ Image /page/10/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 is to the left of the word. Below "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin line. 0(k) Su Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 8 of 14 #### RSV - Simplexa™ Flu A/B & RSV Direct Gen 1.0 (K120413) to Simplexa™ Flu A/B & RSV Direct Gen 2.0 | Simplexa™ Flu A/B & RSV Direct (Gen 2.0) RSV Result | Simplexa™ Flu A/B & RSV Direct (Gen 1.0) RSV Result | | Grand Total | | %Agreement | 95% CI | |--------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------|--------------|----------------|-----|-----------------|----------------| | | Detected | Not Detected | | | | | | Detected | 45 | 9d | 54 | PPA | 97.8% (45/46) | 88.7% to 99.6% | | Not Detected | 1e | 210 | 211 | NPA | 95.9% (210/219) | 92.4% to 97.8% | | Grand Total | 46 | 219 | 265 | | | | | d3/9 discrepant (K120413 – Negative and K142365 – Positive) samples were positive for RSV on another FDA cleared NAT. | | | | | | | | e1/1 discrepant (K120413 – Positive and K142365 – Negative) sample was negative for RSV on another FDA cleared NAT. | | | | | | | PPA: Positive Percent Agreement, NPA: Negative Percent Agreement #### RSV - Simplexa™ Flu A/B & RSV Direct Gen 2.0 to Simplexa™ Flu A/B & RSV Direct Gen 2.1 (K142365) | Simplexa™ Flu A/B & RSV Direct (Gen 2.1) RSV Result | Simplexa™ Flu A/B & RSV Direct (Gen 2.0) RSV Result | | Grand Total | | %Agreement | 95% CI | |-----------------------------------------------------------------|--------------------------------------------------------|--------------|----------------|-----|------------------|-----------------| | | Detected | Not Detected | | | | | | Detected | 55 | 0 | 55 | PPA | 100.0% (55/55) | 93.5% to 100.0% | | Not Detected | 0 | 210 | 210 | NPA | 100.0% (210/210) | 98.2% to 100.0% | | Grand Total | 55 | 210 | 265 | | | | PPA: Positive Percent Agreement, NPA: Negative Percent Agreement #### ANALYTICAL REACTIVITY/ CROSS REACTIVITY #### Analytical Reactivity - Simplexa™ Flu A/B & RSV Direct (Gen 2.1) Different strains of influenza A including H1, H3 and H5 subtypes, influenza B and RSV including A and B subtypes were evaluated. The most recent strains and geographically diverse strains were chosen. Quantified viral material was spiked into negative swab matrix at a single dilution at the concentrations below. Each was assayed in triplicate. Ct values obtained during testing indicate all viral strains were the LoD. All strains tested were appropriately detected. | Organism | Concentration Tested | Result | |---------------------------------------------|----------------------|----------------| | Influenza A/Taiwan/42/06 H1N1 | 100 TCID50/mL | Flu A Detected | | Influenza A/Anhui/1/2013 (H7N9) | 25,000 EID50/mL | Flu A Detected | | Influenza A/Brisbane/10/07 H3 | 100 TCID50/mL | Flu A Detected | | Influenza A/Brisbane/59/07 H1 | 100 TCID50/mL | Flu A Detected | | Organism | Concentration Tested | Result | | Influenza A/California/12/2012 (H1N1) pdm09 | 100 TCID50/mL | Flu A Detected | | Influenza A/California/7/2009 NYMC x-179-A | 100 IU/mL | Flu A Detected | | Influenza A/Indiana/08/2011 (H3N2)v | 100 TCID50/mL | Flu A Detected | | Influenza A/Minnesota/11/2010 (H3N2)v | 100 CEID50/mL | Flu A Detected | | Influenza A/New Caledonia/20/99 H1N1 | 100 TCID50/mL | Flu A Detected | | Influenza A/Ohio/02/2012 (H3N2) | 200 CEID50/mL | Flu A Detected | | Influenza A/Port Chalmers/1/73 H3N2 | 100 TCID50/mL | Flu A Detected | | Influenza A/PR/8/34 H1N1 | 100 TCID50/mL | Flu A Detected | | Influenza A/Solomon Island/03/06 | 100 TCID50/mL | Flu A Detected | | Influenza A/Swine NY/02/2009 H1N1 | 100 TCID50/mL | Flu A Detected | | Influenza A/Swine/1976/31 H1N1 | 100 TCID50/mL | Flu A Detected | | Influenza A/Swine/lowa/15/30 H1N1 | 100 TCID50/mL | Flu A Detected | | Influenza A/Texas/50/2012 (H3N2) | 100 TCID50/mL | Flu A Detected | | Influenza A/Wisconsin/67/05 H3 | 100 TCID50/mL | Flu A Detected | | Influenza A/WS/33 H1N1 | 100 TCID50/mL | Flu A Detected | | Influenza B/Allen/45 | 100 TCID50/mL | Flu B Detected | | Influenza B/Brisbane/60/2008 | 100 CEID50/mL | Flu B Detected | | Influenza B/Florida/02/2006 | 100 TCID50/mL | Flu B Detected | | Influenza B/Florida/04/2006 | 100 TCID50/mL | Flu B Detected | | Influenza B/Florida/07/04 | 100 TCID50/mL | Flu B Detected | | Influenza B/Hong Kong/5/72 | 100 TCID50/mL | Flu B Detected | | Influenza B/Lee/40 | 100 TCID50/mL | Flu B Detected | | Influenza B/Maryland/1/59 | 100 TCID50/mL | Flu B Detected | | Influenza B/Panama/45/90 | 100 TCID50/mL | Flu B Detected | | Influenza B/Taiwan/2/62 | 100 TCID50/mL | Flu B Detected | | Influenza B/Wisconsin/01/2010 | 100 CEID50/mL | Flu B Detected | | RSV-A Long | 100 TCID50/mL | RSV Detected | | RSV-B 9320 | 100 TCID50/mL | RSV Detected | | Organism | Concentration Tested | Result | | RSV-B Wash/18537/62 | 100 TCID50/mL | RSV Detected | | RSV B WV/14617/85 | 100 TCID50/mL | RSV Detected | {11}------------------------------------------------ Image /page/11/Picture/0 description: The image shows the logo for Focus Diagnostics. The word "FOCUS" is in large, bold, black letters. To the left of the word is a black curved shape. Below the word "FOCUS" is the word "Diagnostics" in a smaller font with a line above it. # K142365 510(k) Summary Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa"" Flu A/B & RSV Positive Control Pack [REE] MOL2660 Prepared Date: December 2, 2014 Page 9 of 14 {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, black shape is positioned to the left of the word "FOCUS", resembling a stylized check mark or swoosh. A horizontal line is placed above the word "Diagnostics". 510(k) Summary Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack |REF MOL2660 Prepared Date: December 2, 2014 Page 10 of 14 NOTE: Although this test has been shown to detect novel avian influenza A (H7N9) and H3N2v cultured viruses, the performance characteristics of this device with clinical specimens that are positive for novel avian influenza A (H7N9) and H3N2v influenza viruses have not been established. #### Cross Reactivity (Analytical Specificity) - Simplexa™ Flu A/B & RSV Direct (Gen 2.1) Thirty-two (32) organisms were tested at clinically relevant concentrations to evaluate cross-reactivity of the Simplexa™ Flu A/B & RSV Direct. Three (3) instruments were used and fifteen (15) experimental runs across two (2) days by a single (1) operator were performed. Each of the organisms was spiked into a negative matrix and tested in triplicate (3). Baseline negative matrix was tested in five (5) replicates. No cross reactivity was observed. | | Tested | | %Detection (# Detected / # Total) | | | | | |----------------------------------|-----------------------|---|-----------------------------------|-------------|--------------|-------------|--| | Organism | Conc. | N | Flu A (FAM) | Flu B (JOE) | RSV (CFR610) | IC (Q670) | | | Baseline | N/A | 5 | 0.0%(0/5) | 0.0%(0/5) | 0.0%(0/5) | 100.0%(5/5) | | | Adenovirus 1 | 4.17E+05 TCID50/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Adenovirus 7A | 5.37E+05 TCID50/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Bordetella pertussis A639 | 1.88E+06 cfu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Chlamydia pneumoniae | 1.00E+06 IFU/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Cytomegalovirus (CMV) | 1.04E+05 U/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Coronavirus 229E | 5.89E+05 TCID50/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Coronavirus OC43 | 1.95E+05 TCID50/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Corynebacterium diphtheriae | 4.00E+06 cfu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Enterovirus Type 71 | 1.10E+05 TCID50/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Epstein-Barr Virus (EBV) | 1.10E+05 copies/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Escherichia coli 0157:H7 | 1.10E+06 cfu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Haemophilus influenzae | 1.41E+06 cfu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Lactobacillus plantarum, 17-5 | 7.97E+06 cfu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Legionella longbeachae | 8.63E+06 cfu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Measles | 1.95E+05 TCID50/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Metapneumovirus 9 | 1.58E+05 TCID50/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | | Moraxella catarrhalis, NE1 | 1.49E+06 cfu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | {13}------------------------------------------------ Image /page/13/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in bold, sans-serif font. Above and to the left of the word "FOCUS" is a curved, swooping shape that resembles a check mark. Below the word "FOCUS" is the word "Diagnostics" in a smaller, sans-serif font, underlined with a thin line. 510(k) Summary Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 11 of 14 | | | N | %Detection (# Detected / # Total) | | | | |------------------------------------------------|-----------------------|---|-----------------------------------|-------------|--------------|-------------| | Organism | Tested Conc. | | Flu A (FAM) | Flu B (JOE) | RSV (CFR610) | IC (Q670) | | Mumps | 8.51E+05 TCID50/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | Mycobacterium tuberculosis (genomic DNA) | 6.54E+06 c/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | Mycoplasma pneumoniae, M129 | 3.16E+06 ccu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | Neisseria elongata | 2.05E+06 cfu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | Neisseria meningitides | 7.07E+06 cfu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | Parainfluenza 1 | 1.15E+05 TCID50/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | Parainfluenza 2 | 3.80E+05 IU/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | Parainfluenza 3 | 1.95E+05 TCID50/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | Pseudomonas aeruginosa | 3.93E+06 cfu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | Rhinovirus 1A | 1.26E+05 TCID50/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | Staphylococcus aureus, COL | 1.42E+06 cfu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | Staphylococcus epidermidis | 9.23E+06 cfu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | Streptococcus pneumoniae | 9.20E+06 cfu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | Streptococcus pyogenes, M1 | 1.36E+06 cfu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | Streptococcus salivarius | 2.12E+06 cfu/mL | 3 | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | ### INTERFERENCE - Simplexa™ Flu A/B & RSV Direct (Gen 2.1) The performance of this assay was evaluated with potentially interfering substances that may be present in nasopharyngeal swabs at the concentrations indicated in the table below. The potentially interfering substances were evaluated in a contrived sample that contained influenza B and RSV. All strains were tested at two to four times the LoD. There was no evidence of interference caused by the substances tested. | Substance | Concentration Tested | Qualitative Result for each Channel | | | | |-----------------------|----------------------|-------------------------------------|---------------|---------------|---------------| | | | Flu A (FAM) | Flu B (Joe) | RSV (CFR610) | RNA IC (Q670) | | Baseline | None | 100.0%(15/15) | 100.0%(15/15) | 100.0%(15/15) | 100.0%(15/15) | | Afrin Nasal Spray | 15% (v/v) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | | Oseltamivir phosphate | 1 $\mu$ M | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | | Blood | 2% (v/v) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | | Zicam Nasal Gel | 5% (v/v) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | | Tobramycin | 4 $\mu$ g/mL | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | | Mupirocin | 6.6 mg/mL | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | {14}------------------------------------------------ Image /page/14/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, thinner, black letters with a line above it. 510(k) Summary Simplexa™ Flu A/B & RSV Direct REF MOL2650 Simplexa" Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 12 of 14 | Substance | Concentration Tested | Qualitative Result for each Channel | | | | |------------------------------------------|-------------------------|-------------------------------------|-------------|--------------|---------------| | | | Flu A (FAM) | Flu B (Joe) | RSV (CFR610) | RNA IC (Q670) | | Purified Mucin Protein | 60 µg/mL | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | | Beconase AQ | 5% (v/v) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | | Nasal Corticosteroid - Fluticasone | 5% (v/v) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | | Relenza Antiviral Drug - Zanamivir | 3.3 mg/mL | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | | NTC | NA | 0.0%(0/3) | 0.0%(0/3) | 0.0%(0/3) | 100.0%(3/3) | | Positive Control | NA | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | 100.0%(3/3) | #### ANALYTICAL SENSITIVITY/LIMIT OF DETECTION Three (3) strains of influenza A, two (2) strains of influenza B and two (2) strains of RSV were tested to determine the limit of detection (LoD) for the Simplexa™ Flu A/B & RSV Direct. Four concentrations per virus were tested in triplicate during the screening phase to determine the tentative LoD. The lowest concentration with all replicates detected during the screening phase was tested in thirty two (32) replicates to confirm detection at that concentration. The LoD is determined if at least 31/32 (≥95.0%) replicates are detected. The following table shows the comparison of the limit of detection between the original Simplexa™ Flu A/B & RSV Direct (Gen 1.0) and the modified Simplexa™ Flu A/B & RSV Direct (Gen 2.0 and Gen 2.1). | Virus | Gen 1.0 (TCID50/mL) | Gen 2.0 (TCID50/mL) | Gen 2.1 (TCID50/mL) | |-------------------------------------|------------------------|------------------------|------------------------| | Influenza A/ Hong Kong/8/68 (H3N2) | 10 | 0.1 | 0.1 | | Influenza A/PR/8/34 (H1N1) | 0.005 | 0.05 | 0.05 | | Influenza A/Swine NY/02/2009 (H1N1) | 0.1 | 1 | 1 | | Influenza B/Great Lakes/1739/54 | 2 | 20 | 20 | | Influenza B/Malaysia/2506/2004 | 20 | 0.5 | 1 | | RSV-A2 | 1 | 2 | 2 | | RSV B CH93-18 (18) | 3 | 2 | 2 | {15}------------------------------------------------ Image /page/15/Picture/0 description: The image shows the logo for Focus Diagnostics. The logo consists of the word "FOCUS" in bold, sans-serif font, with a curved, swooping shape above it. Below the word "FOCUS" is the word "Diagnostics" in a smaller, less bold font. The logo is in black and white. ### 510(k) Summary Simplexa" Flu A/B & RSV Direct REF MOL2650 Simplexa™ Flu A/B & RSV Positive Control Pack |REF|MOL2660 Prepared Date: December 2, 2014 Page 13 of 14 142365 ### PRECISION - Simplexa™ Flu A/B & RSV Direct (Gen 2.1) The reaction mix (RM) lot to lot reproducibility was evaluated for Simplexa™ Flu A/B & RSV Direct (Gen 2.1) using three (3) Reaction Mix lots and sample panel which includes Low (~1.5 X LoD) and Moderate (~4 X LoD) positive samples for each of the three targets (Flu A, Flu B and RSV) and Positive Control. The low and moderate positive samples of each target were prepared by spiking the viral strain into negative swab matrix. Each sample panel member was tested in duplicate (2) for each Reaction Mix lot in each run, two (2) runs per day for total of three (3) days using a single (1) 3M Integrated Cycler and a single (1) operator. The study produced at least thirty-six (36) replicates for each sample panel member. The Flu B moderate positive sample was tested in forty-one (41) replicates. | Quantitative Summary of Reaction Mix Inter-Lot Reproducibility | | | | | | | | | | | | | | |----------------------------------------------------------------|-------------------------------|----|------------|-----------|-----|-----------|-----|-----------|-----|-------------------|-----|-------|-----| | Analyte (Channel) | Sample Panel Member | N | Mean Ct | Inter-Day | | Inter-Run | | Inter-Lot | | Intra- Run/Lot | | Total | | | | | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV | | Flu A (FAM)…