K Number

Device Name

ASI Evolution

Manufacturer

Arlington Scientific, Inc. (ASI)

Date Cleared

2018-06-14

(227 days)

Product Code

GMQ GMO

Regulation Number

866.3820

Intended Use

The ASI Automated RPR (rapid plasma reagin) Test for Syphilis, for use on the ASI Evolution Automated Syphilis Analyzer, is a qualitative nontreponemal flocculation test for the detection of reagin antibodies in human serum and plasma as a screening test for serological evidence of syphilis. All reactive RPR test samples should be further tested with a treponemal test. The ASI Automated RPR Test for Syphilis for use on the ASI Evolution produces only a reactive or non-reactive result and does not report RPR titers for reactive samples. The ASI Automated RPR Test for Syphilis is for professional use only. The test is intended to be used for in vitro diagnostic testing. The ASI Evolution is intended to be used as a fully automated analyzer to objectively interpret the results of the ASI automated RPR test for syphilis. The ASI Evolution is designed to provide standardized test interpretation and to provide for storage, retrieval, and transmittal of the test results. It is intended to be acquired, possessed and used only by health care professionals. The ASI Evolution analyzer, in conjunction with the ASI Automated RPR Test is intended to be used for in vitro diagnostic testing. The ASI Automated RPR Test for Syphilis for use on the ASI Evolution produces only a reactive or non-reactive result and does not report RPR titers for reactive samples.

Device Description

The ASI Evolution is an integrated digital particle analyzer designed to objectively interpret certain slide agglutination tests manufactured by Arlington Scientific, Inc. (ASI). The ASI Evolution fully automates the sample and reagent handling steps of the test procedure. Qualitative tests are performed by laboratory professionals who use the ASI Evolution to provide standardized test interpretation using criteria that define reactive and nonreactive aqqlutination reactions. The ASI Evolution employs a camera that uses light reflectance to create a highly sensitive and high-resolution image of the agglutination immunoassay. This image is then analyzed by the proprietary software algorithm to interpret the agglutination pattern. The ASI Evolution further provides tools that enable the creation, storage, retrieval and transmittal of the test results. The ASI Automated RPR Test for Syphilis reagents include the following: CARBON ANTIGEN - 0.003% cardiolipin, 0.020-0.022% lecithin, 0.09% cholesterol, charcoal (activated) as visual enhancer, phosphate buffer, 0.1% sodium azide as preservative and stabilizers. CONTROLS (REACTIVE, WEAK REACTIVE, NONREACTIVE) - Human serum or defibrinated plasma (liquid), with 0.1% sodium azide as preservative. Reagents have two-year expiration dating from date of manufacture. The specific expiration date is located on the label on the vial.

More Information

Contact

Type

Center

Panel

Date Received

Product Code

Subsequent Product Codes

Applicant Name (Manufacturer)

Device Name

Decision

Street 1

Street 2

City

State

Country Code

ZIP

Postal Code

Class Advise Committee

SSP Indicator

Third Party Reviewed

Expedited Review

Predicate Devices

K851504

Reference Devices

K851504

AI/ML (Artificial Intelligence / Machine Learning)

Unknown
The document mentions a "proprietary software algorithm" that analyzes images to interpret agglutination patterns. While this involves image analysis, it doesn't explicitly state or provide details suggesting the use of AI or ML techniques like deep learning or neural networks. The description of the algorithm is too general to confirm the presence of AI/ML.

Therapeutic

This device is an in vitro diagnostic (IVD) test used to screen for syphilis by detecting reagin antibodies in human serum and plasma. It is used for diagnostic purposes and does not provide therapy or treatment.

Diagnostic

Yes

The text explicitly states: "The ASI Automated RPR Test for Syphilis...is a qualitative nontreponemal flocculation test for the detection of reagin antibodies in human serum and plasma as a screening test for serological evidence of syphilis." It also mentions "The test is intended to be used for in vitro diagnostic testing" multiple times.

SaMD (Software as a Medical Device)

The device description clearly states that the ASI Evolution is an "integrated digital particle analyzer" and employs a "camera" to create images. This indicates the presence of hardware components beyond just software.

IVD (In Vitro Diagnostic)

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

The Intended Use section explicitly states: "The test is intended to be used for in vitro diagnostic testing." and "The ASI Evolution analyzer, in conjunction with the ASI Automated RPR Test is intended to be used for in vitro diagnostic testing."

The Device Description also describes the reagents used in the test, which are components of an in vitro diagnostic test.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The ASI Automated RPR Test for Syphilis is for professional use only. The test is intended to be used for in vitro diagnostic testing.

The ASI Evolution is intended to be used as a fully automated analyzer to objectively interpret the results of the ASI automated RPR test for syphilis. The ASI Evolution is designed to provide standardized test interpretation and to provide for storage, retrieval, and transmittal of the test results. It is intended to be acquired, possessed and used only by health care professionals. The ASI Evolution analyzer, in conjunction with the ASI Automated RPR Test is intended to be used for in vitro diagnostic testing. The ASI Automated RPR Test for Syphilis for use on the ASI Evolution produces only a reactive or non-reactive result and does not report RPR titers for reactive samples.

Product codes

GMO

Device Description

The ASI Evolution further provides tools that enable the creation, storage, retrieval and transmittal of the test results.

The ASI Automated RPR Test for Syphilis reagents include the following:

CARBON ANTIGEN - 0.003% cardiolipin, 0.020-0.022% lecithin, 0.09% cholesterol, charcoal (activated) as visual enhancer, phosphate buffer, 0.1% sodium azide as preservative and stabilizers.

CONTROLS (REACTIVE, WEAK REACTIVE, NONREACTIVE) - Human serum or defibrinated plasma (liquid), with 0.1% sodium azide as preservative.

Reagents have two-year expiration dating from date of manufacture. The specific expiration date is located on the label on the vial.

Mentions image processing

Yes

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Light reflectance

Anatomical Site

Not Found

Indicated Patient Age Range

The age of these women ranged in age from 15 to 41 years old. (with a median age of 28 years old).
The women ranged in age from 18 to 40 years old (with a median age of 28 years old).

Intended User / Care Setting

health care professionals / in vitro diagnostic testing

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

Clinical Data: A comparison of the digital interpretation of the results of testing samples with the ASI Automated RPR Test for Syphilis using the ASI Evolution and the ASiManager-AT by trained laboratory professionals was conducted to show substantial equivalence.
The ASI Evolution was evaluated for equivalence, in its pattern of reactivity, against the ASiManager-AT.

Prospective Serum Sample Testing: A total of 1,068 individual prospective samples, with identifiers removed, were collected at two different Departments of Public Health Labs and tested by the ASI Evolution in comparison with the ASiManager-AT at each of the facilities. Reactive, Weak Reactive and Nonreactive controls were run on each day of testing.
Retrospective Serum Sample Testing: A total of 1,013 individual retrospective samples (10 serum, 1003 plasma), with identifiers removed, were collected from various reference labs and serum and plasma vendors from across the United States and tested by the ASI Evolution in comparison with the ASiManager-AT in-house. Reactive, Weak Reactive and Nonreactive controls were run on each day of testing.
ASI Evolution Characterized Specimen Testing: Testing was conducted at Arlington Scientific, Inc. Site C. Carbon antigen lot used Lot CA5K01RBA. Each sample was tested by an operator with experience in performing the ASI Automated RPR Test for Syphilis and operating the ASI Evolution. ASI Evolution unit 5800-0102 (#1) used for testing. Expected results are based on known clinical diagnosis. All samples were serum.
Performance with Samples from Pregnant Women: Samples were collected from 250 pregnant women and tested on the ASI Evolution for reactivity. These samples were also tested using the ASiManager-AT. All samples were nonreactive for syphilis when tested on the ASI Evolution. Samples were collected from 30 pregnant women that also had been diagnosed as having syphilis and were tested on the ASI Evolution for reactivity. All samples were reactive for syphilis when tested on the ASI Evolution. All the samples were serum.

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

Clinical Data:

Prospective Serum Sample Testing: 1,068 samples.
- ASI Automated RPR Test for Syphilis on the ASI Evolution Reactive Result: 114
- ASI Automated RPR Test for Syphilis on the ASI Evolution Nonreactive Result: 1
- ASI RPR Card Test for Syphilis on the ASiManager-AT Reactive Result: 1
- ASI RPR Card Test for Syphilis on the ASiManager-AT Nonreactive Result: 952
- Positive agreement of the ASI Evolution: 114/(114 + 1) = 99.13% (95% Cl = 95.25% - 99.98%)
- Negative agreement of the ASI Evolution: 952/(952 + 1) = 99.9% (95% Cl = 99.42% - 100%)
Retrospective Serum Sample Testing: 10 samples (serum), 1003 samples (plasma).
- Serum: Reactive: 7, Nonreactive: 3.
  - Serum positive agreement: 7/(7 + 0) = 100% (95% Cl = 59.04% - 100%)
  - Serum negative agreement: 3/(3 + 0) = 100% (95% Cl = 29.24% - 100%)
- Plasma: Reactive: 10, Nonreactive: 993.
  - Sodium Citrate positive agreement: 10/(10 + 0) = 100% (95% Cl = 69.15% - 100%)
  - Sodium Citrate negative agreement: 993/(993 + 0) = 100% (95% Cl = 99.63% - 100%)
ASI Evolution Characterized Specimen Testing: 143 samples (serum).
- Primary Treated: 25 samples, 100% Agreement (86.28 – 100%)
- Primary Untreated: 18 samples, 100% Agreement (81.47 – 100%)
- Secondary Treated: 25 samples, 100% Agreement (86.28 – 100%)
- Secondary Untreated: 25 samples, 100% Agreement (86.28 – 100%)
- Latent Treated: 25 samples, 100% Agreement (86.28 – 100%)
- Latent Untreated: 25 samples, 100% Agreement (86.28 – 100%)
Performance with Samples from Pregnant Women: 280 samples (serum).
- Reactive result (ASI Evolution) vs Reactive result (ASiManager-AT): 30
- Nonreactive result (ASI Evolution) vs Nonreactive result (ASiManager-AT): 250
- All 280 samples showed 100% agreement between ASI Evolution and ASiManager-AT.
- Conclusion: The positive and negative percent agreement for the ASI Evolution and the ASiManager-AT demonstrate that they have a very similar performance.

ASI Evolution Performance Characteristics:

Precision: 10 samples repeated 192 times each.
- All 10 samples showed 100% agreement with their expected results (Reactive or Nonreactive).
- The data show that the ASI Evolution has a high degree of repeatability.
Reproducibility:
- Study 1: 7 samples, 180 runs each (60/site at 3 sites).
  - All samples (RPR nonreactive 02287, RPR nonreactive N6K14, RPR reactive 1:2 05225B, RPR reactive 1:4 07035, RPR reactive 1:8 05225A, RPR reactive 1:8 R7C21R, RPR reactive 1:16 07117) showed 100% agreement across all sites and total.
  - The data shows a very high degree of reproducibility.
- Study 2: 448 retrospective serum samples (48 nonreactive, 400 reactive) tested at 3 sites.
  - RPR nonreactive: 48 samples showed 100% agreement at each site (48/48) and overall (144/144), with 95% CI of 92.60 - 100 (site) and 97.47 - 100 (total).
  - RPR reactive: 400 samples showed 100% agreement at each site (400/400) and overall (1200/1200), with 95% CI of 99.08 - 100 (site) and 99.69 - 100 (total).
Cross Reactivity/Interfering Substances:
- Study included various categories (ANA (+), ASO (+), CRP (+), Infectious Mono (+), RF (+), Rubella (+), Lyme's (+), HIV (+), Pregnancy (+), Bilirubin, Hemoglobin, Triglycerides).
- All samples (total 430) showed expected results (Reactive or Nonreactive), i.e., 100% agreement for each category.
- The study showed no interference.
Carry-Over: Conducted alternating a high reactive sample and a nonreactive sample.
- 24 reactive and 24 nonreactive expected results. All results matched expectations (24 reactive, 24 nonreactive).
- This data demonstrates that all testing results were as expected and there was no evidence of contamination or carry-over.
On-board Stability Testing:
- 4 samples (nonreactive, low reactive, moderate reactive, high reactive) tested over 5 days by 2 operators.
- All samples showed 100% agreement with expected results (3/3 trials daily).
- The carbon antigen reagent is stable on the instrument for up to 8 hours.
Frozen vs Refrigerated Testing:
- Various samples with different titers (Not Reactive, 1:1, 1:2, 1:4, 1:8, 1:16)
- All samples gave the expected results for both fresh and frozen conditions.

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

Positive agreement: 99.13%
Negative agreement: 99.9%
Repeatability: 100%
Reproducibility: 100%
Accuracy/Agreement: 100% for characterized specimens, pregnant women, interference, carry-over, and stability studies.

Predicate Device(s)

K851504

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 866.3820

Treponema pallidum nontreponemal test reagents.(a)
Identification. Treponema pallidum nontreponemal test reagents are devices that consist of antigens derived from nontreponemal sources (sources not directly associated with treponemal organisms) and control sera (standardized sera with which test results are compared) used in serological tests to identify reagin, an antibody-like agent, which is produced from the reaction of treponema microorganisms with body tissues. The identification aids in the diagnosis of syphilis caused by microorganisms belonging to the genusTreponema and provides epidemiological information on syphilis.(b)
Classification. Class II (performance standards).

Summary

Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, with the letters "FDA" in a blue square, followed by the words "U.S. FOOD & DRUG" in blue, and then the word "ADMINISTRATION" in a smaller font below.

June 14, 2018

Arlington Scientific, Inc. (ASI) David Binks COO/VP Regulatory 1840 North Technology Dr. Springville, Utah 84663

Re: K173376

Trade/Device Name: ASI Automated RPR Test for Syphilis ASI Evolution Regulation Number: 21 CFR 866.3820 Regulation Name: Treponema pallidum nontreponemal test reagents Regulatory Class: Class II Product Code: GMO Dated: June 7, 2018 Received: June 8, 2018

Dear David Binks:

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 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set 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.

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 comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely.

Steven R. Gitterman -S for

Uwe Scherf, Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

Indications for Use

510(k) Number (if known)

K173376

Device Name

ASI Automated RPR Test for Syphilis ASI Evolution

Indications for Use (Describe)

The ASI Automated RPR Test for Syphilis is for professional use only. The test is intended to be used for in vitro diagnostic testing.

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

☑ Prescription Use (Part 21 CFR 801 Subpart D)	☐ Over-The-Counter Use (21 CFR 801 Subpart C)
--------------------------------------------------------------------------------------------	-----------------------------------------------------------------------

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Arlington Scientific, Inc.

5.0 510(k) Summary

5.1 Preparation Date: 04/23/2018

Submitted By

David Binks MT (ASCP), MBA COO/Vice President Regulatory Arlington Scientific, Inc. 1840 North Technology Dr. Springville, UT 84663 Phone 801-489-8911 / Fax 801-489-5552

5.2 Trade Name - ASI Automated RPR Test for Syphilis ASI Evolution
Regulation section: (21 CFR 866.3820) Treponema pallidum test reagents

Classification: Class II

Product Code: GMQ

Panel: Microbiology

5.3 Predicate Device(s) – ASI RPR Card Test for Syphilis on the ASiManager-AT (K851504)

Device Similarities and Differences
Item	ASI Evolution	ASIManager-AT
Intended Use	The ASI Automated RPR (rapid plasma
reagin) Test for Syphilis, for use on the ASI
Evolution Automated Syphilis Analyzer, is
a qualitative nontreponemal flocculation
test for the detection of reagin antibodies in
human serum and plasma as a screening
test for serological evidence of syphilis. All
reactive RPR test samples should be
further tested with a treponemal test. The
ASI Automated RPR Test for Syphilis for
use on the ASI Evolution produces only a
reactive or non-reactive result and does
not report RPR titers for reactive samples.
The ASI Automated RPR Test for Syphilis
is for professional use only. The test is
intended to be used for in vitro diagnostic
testing.
The ASI Evolution is intended to be used
as a fully automated analyzer to objectively	The ASI RPR (rapid plasma
reagin)
Card Test for Syphilis is a
qualitative and semiquantitative
nontreponemal flocculation test for
the detection of reagin antibodies
in human serum and plasma as a
screening test for serological
evidence of syphilis. This test is
also intended for use in screening
blood donors and cadaveric (non-
heart beating) donor specimens for
tissue donation when the test is
read and interpreted with the
ASIManager-AT. The ASI RPR
Card Test for Syphilis is for
professional use only.
	interpret the results of the ASI automated
RPR test for syphilis. The ASI Evolution is
designed to provide standardized test
interpretation and to provide for storage,
retrieval, and transmittal of the test results.
It is intended to be acquired, possessed
and used only by health care
professionals. The ASI Evolution analyzer,
in conjunction with the ASI Automated
RPR Test is intended to be used for in vitro
diagnostic testing. The ASI Automated
RPR Test for Syphilis for use on the ASI
Evolution produces only a reactive or non-
reactive result and does not report RPR
titers for reactive samples.	The ASiManager-AT is intended to
be used as an integrated digital
particle analyzer to objectively
interpret the results of the ASI
RPR Card Test for Syphilis. The
ASiManager-AT is designed to
provide standardized test
interpretation and to provide for
storage, retrieval, and transmittal
of the test results. It is intended to
be acquired, possessed and used
only by health care professionals.
The ASiManager-AT is intended to
be used for in vitro diagnostics,
blood donor and cadaveric (non-
heart beating) donor screening.
Technology
Instruments	The ASI Evolution is an integrated digital
particle analyzer designed to objectively
interpret certain agglutination tests
manufactured by Arlington Scientific, Inc.
(ASI). The ASI Evolution fully automates
the sample and reagent handling steps of
the test procedure. Laboratory
professionals use the ASI Evolution to
provide standardized test interpretation
using criteria that define reactive and
nonreactive agglutination reactions. The
same proprietary interpretive algorithm
used in the predicate device (ASiManager-
AT) is used in the ASI Evolution.	The ASiManager-AT is an
integrated digital particle analyzer
designed to objectively interpret
certain slide agglutination tests
manufactured by Arlington
Scientific, Inc. (ASI). Qualitative
and semiquantitative tests are
performed by laboratory
professionals who use the
ASiManager-AT to provide
standardized test interpretation
using criteria that define reactive
and nonreactive agglutination
reactions.
	The ASI Evolution employs a camera to
create a highly sensitive and high-
resolution image of the agglutination
immunoassay. This image is then
analyzed by the proprietary software
algorithm to interpret the agglutination
pattern.
The ASI Evolution further provides tools
that enable the creation, storage, retrieval
and transmittal of the test results.	The ASiManager-AT employs a
camera to create a highly sensitive
and high-resolution image of the
agglutination immunoassay. This
image is then analyzed by the
proprietary software algorithm to
interpret the agglutination pattern.
The ASiManager-AT further
provides tools that enable the
creation, storage, retrieval and
transmittal of the test results.
Technology
Reagents	Flocculation Test	Same
Antigen	ASI RPR Carbon Antigen	Same
Reported Results	Reactive, Nonreactive	Same
Interpretation	Automated	Same
Sample Processing	Automated	Manual
Reagent Volume
used per Sample	110 $μ$ l	105 $μ$ l
Sample Type	Serum or Plasma	Same
Controls	Reactive, Weak Reactive, Nonreactive	Same
Test card	48 well plastic test plate	30, 15, or 10 well plastic coated
test card
Target Population	Used for in vitro diagnostic testing and	Used for in vitro diagnostics
testing, blood donor and cadaveric

Device Similarities and Differences

Arlington Scientific, Inc.

Arlington Scientific, Inc.

		(non-heart beating) donor screening.
--	--	--------------------------------------

5.4 Device Description – The ASI Evolution is an integrated digital particle analyzer designed to objectively interpret certain slide agglutination tests manufactured by Arlington Scientific, Inc. (ASI). The ASI Evolution fully automates the sample and reagent handling steps of the test procedure. Qualitative tests are performed by laboratory professionals who use the ASI Evolution to provide standardized test interpretation using criteria that define reactive and nonreactive aqqlutination reactions.
The ASI Evolution employs a camera that uses light reflectance to create a highly sensitive and high-resolution image of the agglutination immunoassay. This image is then analyzed by the proprietary software algorithm to interpret the agglutination pattern.

The ASI Evolution further provides tools that enable the creation, storage, retrieval and transmittal of the test results.

The ASI Automated RPR Test for Syphilis reagents include the following:

CARBON ANTIGEN - 0.003% cardiolipin, 0.020-0.022% lecithin, 0.09% cholesterol, charcoal (activated) as visual enhancer, phosphate buffer, 0.1% sodium azide as preservative and stabilizers.

CONTROLS (REACTIVE, WEAK REACTIVE, NONREACTIVE) - Human serum or defibrinated plasma (liquid), with 0.1% sodium azide as preservative.

Reagents have two-year expiration dating from date of manufacture. The specific expiration date is located on the label on the vial.

Intended Use – The ASI Evolution is intended to be used as a fully automated analyzer to objectively interpret the results of the ASI Automated RPR Test for Syphilis. The ASI Evolution is designed to provide standardized test interpretation and to provide for storage, retrieval, and transmittal of the test results. It is intended to be acquired, possessed and used only by health care professionals. The ASI Evolution analyzer, in conjunction with the ASI Automated RPR Test for Syphilis is intended to be used for in vitro diagnostic testing. The ASI Automated RPR Test for Syphilis for use on the ASI Evolution produces only a reactive or non-reactive result and does not report RPR titers for reactive samples.

Arlington Scientific, Inc.

Test for Syphilis for use on the ASI Evolution produces only a reactive or nonreactive result and does not report RPR titers for reactive samples. The ASI Automated RPR Test for Syphilis is for professional use only. The test is intended to be used for in vitro diagnostic testing.

5.5 Clinical Data - A comparison of the digital interpretation of the results of testing samples with the ASI Automated RPR Test for Syphilis using the ASI Evolution and the ASiManager-AT by trained laboratory professionals was conducted to show substantial equivalence.
The ASI Evolution was evaluated for equivalence, in its pattern of reactivity, aqainst the ASiManager-AT. A total of 1,068 individual prospective samples, with identifiers removed. were collected at two different Departments of Public Health Labs and tested by the ASI Evolution in comparison with the ASiManager-AT at each of the facilities. Reactive, Weak Reactive and Nonreactive controls were run on each day of testing.

Prospective Serum Sample Testing - 1,068 Samples ASI RPR Card Test for Syphilis on the ASiManager-AT Result ASI Automated Reactive Nonreactive RPR Test for Syphilis on the 114 1 ASI Evolution Reactive Result Nonreactive 1 952

The following data are the results from two testing sites:

Using the data from the prospective performance results above, the positive agreement of the ASI Evolution can be calculated as:

114/(114 + 1) = 99.13% 95% Cl = 95.25% - 99.98%

Using the data from the prospective performance results above, the negative agreement of the ASI Evolution can be calculated as:

952/(952 + 1) = 99.9% 95% Cl = 99.42% - 100%

A total of 1,013 individual retrospective samples, with identifiers removed, were collected from various reference labs and serum and plasma vendors from across the United States and tested by the ASI Evolution in comparison with the ASiManager-AT

Arlington Scientific, Inc.

in-house. Reactive, Weak Reactive and Nonreactive controls were run on each day of testing.

Arlington Scientific, Inc.

Retrospective Serum Sample Testing – 10 Samples
ASI RPR Card Test for Syphilis on the ASiManager-AT Result
	Reactive	Nonreactive
ASI Automated
RPR Test for
Syphilis on the
ASI Evolution
Result	7	0
Nonreactive	0	3

Retrospective Serum Sample Testing – 10 Samples

Serum positive agreement is calculated as:

7/(7 + 0) = 100% 95% Cl = 59.04% - 100%

Serum negative agreement is calculated as:

3/(3 + 0) = 100% 95% Cl = 29.24% - 100%

Retrospective Plasma Sample Testing – 1,003 Samples
ASI Automated
RPR Test for
Syphilis on the
ASI Evolution
Result	ASI RPR Card Test for Syphilis on the ASiManager-AT
Result
	Reactive	Nonreactive
Reactive	10	0
Nonreactive	0	993

Sodium Citrate positive agreement is calculated as:

10/(10 + 0) = 100% 95% Cl = 69.15% - 100%

Sodium Citrate negative agreement is calculated as:

993/(993 + 0) = 100% 95% Cl = 99.63% - 100%

Arlington Scientific, Inc.

	Prospective random samples			Retrospective samples
Site	Plasma	Serum	Total	Serum & Plasma		Total	Grand Total
				Known infected	Known Uninfected
a	0	567	567	0	0	0	567
b	0	501	501	0	0	0	501
c	0	0	0	17	996	1,013	1,013
total	0	1,068	1,068	17	996	1,013	2,081

Distribution of Samples

ASI Evolution Characterized Specimen Testing

Testing was conducted at: .
- o Arlington Scientific, Inc. Site C
Carbon antigen lot used Lot CA5K01RBA ●
Each sample was tested by an operator with experience in performing the ASI ● Automated RPR Test for Syphilis and operating the ASI Evolution.
ASI Evolution unit 5800-0102 (#1) used for testing .
Expected results are based on known clinical diagnosis ●
All samples were serum .

The results of the testing are contained in tables below:

| Clinical

Diagnosis	No. Reactive	% Agreement	95% CI
Primary Treated	25	100%	86.28 – 100%
Primary Untreated	18	100%	81.47 – 100%
Secondary Treated	25	100%	86.28 – 100%
Secondary Untreated	25	100%	86.28 – 100%
Latent Treated	25	100%	86.28 – 100%
Latent Untreated	25	100%	86.28 – 100%

Performance with Samples from Pregnant Women:

Samples were collected from 250 pregnant women and tested on the ASI Evolution for reactivity. These samples were also tested using the ASiManager-AT. The age of these women ranged in age from 15 to 41 years old. (with a median age of 28 years old). All samples were nonreactive for syphilis when tested on the ASI Evolution. Samples were collected from 30 pregnant women that also had been diagnosed as having syphilis and were tested on the ASI Evolution for reactivity. The women ranged in age from 18 to 40 years old (with a median age of 28 years old). All samples were reactive for syphilis when tested on the ASI Evolution. All the samples were serum.

Arlington Scientific, Inc.

Pregnant Women Testing

| ASI Automated
RPR Test for
Syphilis on the
ASI Evolution
Result | ASI RPR Card Test for Syphilis on the ASiManager-AT
Result | |
|-----------------------------------------------------------------------------|---------------------------------------------------------------|-------------|
| | Reactive | Nonreactive |
| Reactive | 30 | 0 |
| Nonreactive | 0 | 250 |

Conclusion:

The positive and negative percent agreement for the ASI Evolution and the ASiManager-AT demonstrate that they have a very similar performance.

ASI Evolution Performance Characteristics

Precision

The interpretation of 10 samples using the ASI Automated RPR Test for Syphilis and the ASI Evolution were evaluated for reactivity. The testing requirements were as follows:

1. All qualitative testing was conducted according to the procedure listed in the package insert.
1. Each qualitative sample was tested 192 times. The number of replicates were chosen since the instrument can hold 4 - 48 well plates for a total of 192 wells. The 192 samples will test each well position.

A total of 10 samples were evaluated to determine repeatability of reactivity. Of the 10 samples:

3 were nonreactive

4- RPR reactive 1:1 titered samples
1- RPR reactive 1:2 titered sample
1- RPR reactive 1:8 titered sample
1- RPR reactive 1:256 titered sample

Each of the 10 samples was repeated 192 times to evaluate the reactivity using the ASI Evolution. An aliquot of the same sample was dispensed into 192 tubes. All 192 tubes were placed into the ASI Evolution and the run was performed. In this manner all 192 wells were tested with the same sample to show well to well and plate to plate repeatability using four test plates.

Arlington Scientific, Inc.

Precision Testing
Sample	Result		% Agreement
	Sample ID	Titer
1	R7C21R	1:8	R	192/192	100%
2	N7D04	NR	NR	192/192	100%
3	11114B	1:1	R	192/192	100%
4	11114C	1:1	R	192/192	100%
5	11114F	1:1	R	192/192	100%
6	02287	NR	NR	192/192	100%
7	08296	1:256	R	192/192	100%
8	11114D	1:1	R	192/192	100%
9	W7E26R	1:2	R	192/192	100%
10	N7H03	NR	NR	192/192	100%

Precision Testing

The data above show that the ASI Evolution has a high degree of repeatability.

Reproducibility

Reproducibility testing was conducted at three sites. The testing consisted of:

. Testing seven (7) samples
- o 2 RPR nonreactive samples
- o 1 RPR reactive 1:2 titered samples
- o 1 RPR reactive 1:4 titered sample
- o 2 RPR reactive 1:8 titered sample
- o 1 RPR reactive 1:16 titered sample
Each sample was run in duplicate within the panel. .
Each sample was tested each day for five non-consecutive days by an operator . with experience in performing the ASI Automated RPR Test for Syphilis and operating the ASI Evolution.
Each sample was tested a second time on each of the days referenced above . separated by approximately 2 hours.
. Testing at each site was performed on the same ASI Evolution.

Arlington Scientific, Inc.

RPR		Site 1		Site 2		Site 3		Total
Sample	Sample

| N | Expected

Result | 95%
Confidence
Interval | %
Expected
Result | 95%
Confidence
Interval | %
Expected
Result | 95%
Confidence
Interval | %
Expected
Result | 95%
Confidence
Interval |
| RPR
nonreactive | 02287 | 180 | 100%
(60/60) | 93.98 - 100 | 100%
(60/60) | 93.98 - 100 | 100%
(60/60) | 93.98 - 100 | 100%
(180/180) | 97.91 - 100 |
| RPR
nonreactive | N6K14 | 180 | 100%
(60/60) | 93.98 - 100 | 100%
(60/60) | 93.98 - 100 | 100%
(60/60) | 93.98 - 100 | 100%
(180/180) | 97.91 - 100 |
| RPR reactive
1:2 | 05225B | 180 | 100%
(60/60) | 93.98 - 100 | 100%
(60/60) | 93.98 - 100 | 100%
(60/60) | 93.98 - 100 | 100%
(180/180 | 97.91 - 100 |
| RPR reactive
1:4 | 07035 | 180 | 100%
(60/60) | 93.98 - 100 | 100%
(60/60) | 93.98 - 100 | 100%
(60/60) | 93.98 - 100 | 100%
(180/180) | 97.91 - 100 |
| RPR reactive
1:8 | 05225A | 180 | 100%
(60/60) | 93.98 - 100 | 100%
(60/60) | 93.98 - 100 | 100%
(60/60) | 93.98 - 100 | 100%
(180/180) | 97.91 - 100 |
| RPR reactive
1:8 | R7C21R | 180 | 100%
(60/60) | 93.98 - 100 | 100%
(60/60) | 93.98 - 100 | 100%
(60/60) | 93.98 - 100 | 100%
(180/180) | 97.91 - 100 |
| RPR reactive
1:16 | 07117 | 180 | 100%
(60/60) | 93.98 - 100 | 100%
(60/60) | 93.98 - 100 | 100%
(60/60) | 93.98 - 100 | 100%
(180/180 | 97.91 - 100 |

The data shows a very high degree of reproducibility.

Another reproducibility study was conducted at the three sites. All qualitative testing was conducted according to the procedure listed in the package insert. The testing consisted of:

. Testing 448 samples
- o 48 RPR nonreactive samples
- o 400 RPR reactive samples
The same samples were tested at all three sites described above for the clinical ● and the previous reproducibility studies.
. All samples were retrospective serum.
Samples, with identifiers removed, were collected from various reference labs ● and serum and plasma vendors from across the United States.
Testing at each site was performed on the same ASI Evolution. .
Results were as follows: ●

RPR		Site 1
Sample	N	Expected
Result	95%
Confidence
Interval	%
Expected
Result	95%
Confidence
Interval	%
Expected
Result	95%
Confidence
Interval	% Expected
Result	95%
Confidence
Interval
RPR
nonreactive	48	100%
(48/48)	92.60 - 100	100%
(48/48)	92.60 - 100	100%
(48/48)	92.60 - 100	100%
(144/144)	97.47 - 100
RPR reactive	400	100%
(400/400)	99.08 - 100	100%
(400/400)	99.08 - 100	100%
(400/400)	99.08 - 100	100%
(1200/1200)	99.69 - 100

Arlington Scientific, Inc.

Cross Reactivity/Interfering Substances

A study was conducted to evaluate potential interference or cross reactivity from different disease conditions. Results are listed below:

Specimen Category	Number of Samples	Expected Result	# of samples with expected result/# of samples tests
ANA (+) Syphilis (-)	3	NR	3/3
ASO (+) Syphilis (-)	2	NR	2/2
CRP (+) Syphilis (-)	2	NR	2/2
Infectious Mono (+) Syphilis (-)	3	NR	3/3
RF (+) Syphilis (-)	12	NR	12/12
Rubella (+) Syphilis (-)	12	NR	12/12
Lyme's (+) Syphilis (-)	12	NR	12/12
HIV (+) Syphilis (-)	50	NR	50/50
HIV (+) Syphilis (+)	24	R	24/24
Pregnancy (+) Syphilis (-)	250	NR	250/250
Pregnancy (+) Syphilis (+)	30	R	30/30
Bilirubin 20 mg/dl	2	NR	2/2
Hemoglobin 10 mg/ml	2	NR	2/2
Triglycerides 1000mg/dl	2	NR	2/2

The positive infectious mono samples were heterophile positive. EBV testing was not conducted.

The study showed no interference.

Carry-Over

A study was conducted to evaluate if contamination of a nonreactive sample due to carryover from an adjacent reactive sample can occur.

Testing was conducted at: ●
- o Arlington Scientific, Inc.
Testing was conducted using two different samples: ●
- RPR reactive 1:64 tittered sample (high reactive) 06237 o
- o RPR nonreactive sample Lot 06127
The same samples were used for all testing.
The same lot of carbon antigen was used - Lot CA7D24R
Each test run was completed each day for five days by an operator with experience in performing the ASI Automated RPR Test for Syphilis and operating the ASI Evolution.
The test consisted of alternating 24 aliquots of the samples listed above in the sample rack and completing a run of 48 tests.

Arlington Scientific, Inc.

Testing was performed on the same ASI Evolution. ●

	Expected Results
ASI Automated
RPR Test for
Syphilis on the
ASI Evolution
Result
	Reactive	Nonreactive
Reactive	24	0
Nonreactive	0	24

Carry-Over Testing

This data demonstrates that all testing results were as expected and there was no evidence of contamination or carry-over.

ASI Evolution On-board Stability Testing

Testing was conducted at: ●
- o Arlington Scientific, Inc. Site C
Testing was conducted using four different samples: .
- RPR nonreactive sample Lot 12313 o
- RPR reactive 1:2 tittered sample (low reactive) LR0646530 O
- RPR reactive 1:16 tittered sample (moderate reactive) LB0482464 o
- RPR reactive 1:64 tittered sample (high reactive) OL0500976 o
The same samples were used for all testing. ●
The same lot of carbon antigen was used Lot CA5K01RBA ●
. Each sample was tested each morning for five days by an operator with experience in performing the ASI Automated RPR Test for Syphilis and operating the ASI Evolution.
. Each sample was tested each afternoon for five days by a different operator with experience in performing the ASI Automated RPR Test for Syphilis and operating the Evolution.
The carbon antigen reagent was left on the ASI Evolution between runs for a total elapsed time of 8 hours.
The carbon antigen was stored at 2-8° C between the afternoon test run and the next morning test run (overnight)
Each of the four samples was run in triplicate on the morning and afternoon . runs.
ASI Evolution unit 5800-0102 (#1) used for testing
All samples were serum. .

The results of the testing are contained in table below:

| Sample | Expected | Results | | | | | %
Agreement |
|------------|----------|---------------|-------|-------|-------|-------|----------------|
| | | ASI Evolution | | | | | |
| Operator 1 | | Day 1 | Day 2 | Day 3 | Day 4 | Day 5 | |
| 12313 | NR | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 100% |
| LR0646530 | R | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 100% |
| LB0482464 | R | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 100% |
| OL0500976 | R | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 100% |
| Operator 2 | | | | | | | |
| 12313 | NR | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 100% |
| LR0646530 | R | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 100% |
| LB0482464 | R | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 100% |
| OL0500976 | R | 3/3 | 3/3 | 3/3 | 3/3 | 3/3 | 100% |

On-board Stability Testing

Using this data the following evaluations can be made:

All testing results were as expected.

The carbon antigen reagent is stable on the instrument for up to 8 hours.

ASI Evolution Frozen vs Refrigerated Testing

Testing was conducted at: .
- o Arlington Scientific, Inc. Site C
Carbon antigen Lot used Lot CA5K01RBA .
Each sample was tested by an operator with experience in performing the ASI ● Automated RPR Test for Syphilis and operating the ASI Evolution.
ASI Evolution unit 5800-0102 (#1) used for testing. .
Each sample was split into two aliquots. One was stored at 2-8° C and the other ● one was frozen. The frozen specimens were allowed to remain frozen for 1 month and then were thawed.
All testing was done on the ASI Evolution. ●
. All specimens were allowed to come to room temperature before testing.

| Sample Titer | N | Fresh Samples
% Expected Result
(# expected
result/N) | Frozen Samples
% Expected Result
(# expected
result/N) |
|--------------|----|----------------------------------------------------------------|-----------------------------------------------------------------|
| Not Reactive | 20 | 100% (20/20) | 100% (20/20) |
| 1:1 | 20 | 100% (20/20) | 100% (20/20) |
| 1:2 | 5 | 100% (5/5) | 100% (5/5) |
| 1:4 | 5 | 100% (5/5) | 100% (5/5) |
| 1:8 | 3 | 100% (3/3) | 100% (3/3) |
| 1:16 | 2 | 100% (2/2) | 100% (2/2) |

Frozen vs. Refrigerated Testing

All samples gave the expected results.