Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The summary describes a traditional in vitro diagnostic (IVD) immunoblot assay that relies on the detection of specific antibodies through biochemical reactions and visual interpretation of bands. There is no mention of any computational analysis, image processing, or algorithms that would suggest the use of AI or ML.

Therapeutic

No.
This device is an in vitro diagnostic (IVD) intended for the qualitative detection of IgG antibodies for Borrelia burgdorferi to aid in the diagnosis of Lyme disease, not for direct therapy.

Diagnostic

Yes

The "Intended Use / Indications for Use" section explicitly states that the kit is "intended to aid in the diagnosis of Lyme disease" and is "For in vitro diagnostic use only."

SaMD (Software as a Medical Device)

No

The device is an in vitro diagnostic (IVD) kit that includes physical components (nitrocellulose membrane with applied proteins, conjugate control, calibrator/serum control, etc.) for performing an immunoblot assay. It is not solely software.

IVD (In Vitro Diagnostic)

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

Here's why:

Explicit Statement: The "Intended Use / Indications for Use" section clearly states: "For in vitro diagnostic use only".
Intended Use: The device is intended for the "in vitro qualitative detection of IgG antibodies to Borrelia burgdorferi in human serum." This involves testing samples taken from the human body outside of the body, which is the definition of in vitro.
Purpose: The purpose is to "aid in the diagnosis of Lyme disease," which is a diagnostic purpose.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The iDart™ Lyme IgG ImmunoBlot Kit is an immunoblot assay intended for the in vitro qualitative detection of IgG antibodies to Borrelia burgdorferi in human serum. The iDart Lyme IgG ImmunoBlot Kit is intended to detect antibodies to LSA and multiple other B. burgdorferi antigens following a modified two-tier test methodology. Positive results from the iDart Lyme IgG ImmunoBlot Kit are supportive evidence for the presence of antibodies and exposure to B. burgdorferi. Negative results do not preclude infection with B. burgdorferi. iDart™ Lyme IgG ImmunoBlot Kit is intended to aid in the diagnosis of Lyme disease and the test kit should only be used on samples from patients with clinical history, signs and symptoms consistent with Lyme disease. The iDart Lyme IgG Immunoblot Kit is not intended as a screen for asymptomatic patients.

Test results are to be used in conjunction with information obtained from the patient's clinical evaluation and other diagnostic procedures.

Product codes

LSR

Device Description

The iDart™ Lyme IgG ImmunoBlot tests are line immunoblot assays. Antigenic proteins specific for Borrelia species that cause Lyme Disease are produced by recombinant DNA technology in Escherichia coli. The purified proteins are then applied as discrete lines on a nitrocellulose membrane along with two control proteins.

The iDart™ Lyme IgG ImmunoBlot Kit contains IgG ImmunoBlot strips and the proteins are applied in the following order: C1 (IgG/IgM - conjugate control), C2 (Protein L - calibrator/serum control), P93, P41, P39, P23, P31, P66, P58, P45, P34, P30, P28, P18 and LSA (a chimeric VISE peptide termed the Lyme Screen Antigen).

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

Not Found

Intended User / Care Setting

For professional use only

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

A total of 768 serum samples were procured from two vendors (Bay Area Lyme Foundation and IGeneX Inc.) and tested at three clinical sites. All samples were blinded, re-coded, and tested at the respective clinical sites as per the instructions for use for the iDart Lyme IgG ImmunoBlot Kit.

Summary of Performance Studies

Reproducibility: The iDart™ Lyme IgG ImmunoBlot Kit was tested in a blind study to evaluate reproducibility across 3 separate sites each with 2 operators over 5 days, 2 runs a day, using a panel of blinded and coded samples of negative, moderate negative, high negative, moderate positive and high positive samples for IgG. It is concluded that there was 100% agreement of all bands among all runs, all days and across 3 sites for the iDart™ Lyme IgG ImmunoBlot Kit.
Analytical Specificity/Interference:
- Analytical Specificity: Tested on 313 apparently healthy individuals from endemic areas (Specificity: 99.36%) and 112 healthy individuals from non-endemic areas (Specificity: 100%).
- Cross-Reactivity Study: Tested serum samples (N=376) from patients with bacterial/viral infections and autoimmune diseases. No cross-reactivity was observed.
- Interference from Endogenous Analytes: Evaluated the effect of Bilirubin, Albumin, Cholesterol, Triglycerides, and Hemoglobin on one positive, one low positive, and one negative Borrelia IgG sample. No interference was observed.
Clinical Studies (Method Comparison with comparators (STTT)):
- Study 1 (Bay Area Lyme Foundation, n=290): PPA 95.00% (95% CI: 76.39% – 99.11%), NPA 86.67% (95% CI: 82.09% – 90.21%).
- Study 2 (IGeneX Inc. Cohort 2, n=248): PPA 95.00% (95% CI: 89.52% – 97.69%), NPA 90.63% (95% CI: 84.33% - 94.56%).
- Study 3 (IGeneX Inc. Cohort 3, n=230): PPA 90.91% (95% CI: 62.27% – 98.38%), NPA 96.80% (95% CI: 93.55% – 98.44%).
Clinical Sensitivity/Specificity (CDC Serum Panel, N=280):
- Overall Sensitivity: 71.11% (compared to STTT 52.22%)
- Overall Specificity: 100%
- Stage I: Sensitivity 58.33% (STTT: 30.00%)
- Stage II: Sensitivity 90.00% (STTT: 90.00%)
- Stage III: Sensitivity 100% (STTT: 100%)
- Healthy Controls: Specificity 100%
- Disease Controls: Specificity 100%
Other Clinical Supportive Data:
- Fresh and Frozen Samples Comparison Study (n=72): Performance of iDart™ Lyme IgG ImmunoBlot Kit for fresh and frozen samples are comparable.
- Antibody Class Specificity (n=10): Demonstrated specificity of the goat anti-human IgG Conjugate.

Key Metrics

Reproducibility: 100% agreement of all bands.
Analytical Specificity (Endemic Area): 99.36%
Analytical Specificity (Non-Endemic Area): 100%
Cross-Reactivity Specificity: 100% (for LSA), 98.67% (for 2+ bands), 100% (for IgG Positive).
Method Comparison (Bay Area Lyme Foundation): PPA: 95.00%, NPA: 86.67%
Method Comparison (IGeneX Inc. Cohort 2): PPA: 95.00%, NPA: 90.63%
Method Comparison (IGeneX Inc. Cohort 3): PPA: 90.91%, NPA: 96.80%
Clinical Sensitivity (Overall - CDC Panel): 71.11%
Clinical Specificity (Overall - CDC Panel): 100%

Predicate Device(s)

K220016

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 866.3830

Treponema pallidum treponemal test reagents.(a)
Identification. Treponema pallidum treponemal test reagents are devices that consist of the antigens, antisera and all control reagents (standardized reagents with which test results are compared) which are derived from treponemal sources and that are used in the fluorescent treponemal antibody absorption test (FTA-ABS), theTreponema pallidum immobilization test (T.P.I.), and other treponemal tests used to identify antibodies toTreponema pallidum directly from infecting treponemal organisms in serum. The identification aids in the diagnosis of syphilis caused by bacteria belonging to the genusTreponema and provides epidemiological information on syphilis.(b)
Classification. Class II (performance standards).

Summary

0

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, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.

ID-FISH Technology, Inc. Jyotsna Shah VP of Research and Development 556 Gibraltar Drive Milpitas, California 95035

August 12, 2024

Re: K233367

Trade/Device Name: iDart Lyme IgG ImmunoBlot Kit Regulation Number: 21 CFR 866.3830 Regulation Name: Treponema Pallidum Treponemal Test Reagents Regulatory Class: Class II Product Code: LSR Dated: July 5, 2024 Received: July 8, 2024

Dear Jyotsna Shah:

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 (the Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database available at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).

1

Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30, Design controls; 21 CFR 820.90, Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review, the QS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).

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 Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Noel J. Gerald -S

Noel J. Gerald, Ph.D. Branch Chief Bacterial Respiratory and Medical Countermeasures Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

2

Indications for Use

510(k) Number (if known) K233367

Device Name iDart™ Lyme IgG ImmunoBlot Kit

Indications for Use (Describe)

The iDart™ Lyme IgG ImmunoBlot Kit is an immunoblot assay intended for the in vitro qualitative detection of IgG antibodies to Borrelia burgdorferi in human serum. The iDart Lyme IgG ImmunoBlot Kit is intended to detect antibodies to LSA and multiple other B. burgdorferi antigens following a modified two-tier test methodology. Positive results from the iDart Lyme IgG ImmunoBlot Kit are supportive evidence for the presence of antibodies and exposure to B. burgdorferi. Negative results do not preclude infection with B. burgdorferi. iDart™ Lyme IgG ImmunoBlot Kit is intended to aid in the diagnosis of Lyme disease and the test kit should only be used on samples from patients with clinical history, signs and symptoms consistent with Lyme disease. The iDart Lyme IgG Immunoblot Kit is not intended as a screen for asymptomatic patients.

Test results are to be used in conjunction with information obtained from the patient's clinical evaluation and other diagnostic procedures.

For in vitro diagnostic use only For professional use only For prescription use only

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)
-------------------------------------------------------------------------------------------------------------------	------------------------------------------------------------------------------------------------------------------

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

510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION SUMMARY

BACKGROUND INFORMATION: l.

A. 510(k) Number

K233367

B. Applicant

ID-FISH Technology, Inc. 556 Gibraltar Drive Milpitas CA 95035 USA

C. Contact Person

Dr. Jyotsna Shah VP of Research and Development regulatory@idfishtechnology.com 1-650-269-8610

D. Date Prepared

July 30, 2024

E. Proprietary and Established Names

iDart™ Lyme IqG ImmunoBlot Kit

F. Regulatory Information

Product Code(s): LSR - Reagents, Borrelia Serological Reagent Classification: Class II Regulation Section: 21 CFR 866.3830 - Treponema Pallidum Treponemal Test Reagents Panel: MI - Microbiology

II. SUBMISSION/DEVICE OVERVIEW:

A. Purpose for Submission:

To obtain a substantial equivalence determination for a new device.

B. Measurand:

lgG antibodies to Borrelia burgdorferi (B. burgdorferi)

C. Type of Test:

ImmunoBlot Assay

4

III. INTENDED USE/INDICATIONS FOR USE:

A. Intended Use(s):

The iDart™ Lyme IgG ImmunoBlot Kit is an immunoblot assay intended for the in vitro qualitative detection of lgG antibodies to Borrelia burgdorferi in human serum. The iDart Lyme IgG ImmunoBlot Kit is intended to detect antibodies to LSA and multiple other B. burgdorferi antigens following a modified two-tier test methodology. Positive results from the iDart Lyme IgG ImmunoBlot Kit are supportive evidence for the presence of antibodies and exposure to B. burgdorferi. Negative results do not preclude infection with B. burgdorferi. iDart™ Lyme IgG lmmunoBlot Kit is intended to aid in the disease and the test kit should only be used on samples from patients with clinical history, signs and symptoms consistent with Lyme disease. The iDart Lyme IgG Immunoblot Kit is not intended as a screen for asymptomatic patients.

Test results are to be used in conjunction with information obtained from the patient's clinical evaluation and other diagnostic procedures.

B. Indication(s) for Use:

iDart™ Lyme IgG ImmunoBlot Kit should be used for diagnosing Lyme Disease in patients with a clinical history, and signs and symptoms consistent with Lyme Disease. It is not intended for use in asymptomatic patients.

C. Special Conditions for Use Statement(s):

For in vitro diagnostic use only For professional use only For prescription use only

IV. DEVICE CHARACTERISTICS:

A. Device Description:

The iDart™ Lyme IgG ImmunoBlot tests are line immunoblot assays. Antigenic proteins specific for Borrelia species that cause Lyme Disease are produced by recombinant DNA technology in Escherichia coli. The purified proteins are then applied as discrete lines on a nitrocellulose membrane along with two control proteins.

The iDart™ Lyme IgG ImmunoBlot Kit contains IgG ImmunoBlot strips and the proteins are applied in the following order: C1 (lgG/lgM - conjugate control), C2 (Protein L - calibrator/serum control), P93, P41, P39, P23, P31, P66, P58, P45, P34, P30, P28, P18 and LSA (a chimeric VISE peptide termed the Lyme Screen Antigen).

B. Principle of procedures:

During the test procedure, diluted human serum is added to the immunoblot strip. Antibodies to Borrelia infection, if present, will bind to antigen lines on the nitrocellulose strips. After removing serum and unbound antibodies by washing, the nitrocellulose strip is incubated with alkaline phosphatase conjugated anti-human IgG antibody for detecting IgG antibodies.

After removing the unbound alkaline phosphatase conjugated antibody by washinq, the antibody complex is visualized as bands by adding the alkaline phosphatase substrate 5-bromo, 4-chloro, 3indolylphosphate (BCIP) and nitro blue tetrazolium (NBT) to form a strong bluish-purple precipitate. The reaction is stopped by washing the nitrocellulose strip with distilled or deionized water. A strip reading guide included in each test kit shows the location of specific antigens in the test strip. Every immunoblot strip has two functional control bands. The test strip is only valid when both control bands are visible after completion of the test. Any band found having a visual intensity equal to or greater than the C2 control band intensity is considered as a significant (positive) band. Depending on the observed bands pattern, one can interpret the presence or absence of Lyme specific IgG antibodies in the patient serum.

5

SUBSTANTIAL EQUIVALENCE INFORMATION: V.

A. Predicate Device Name(s):

Viramed Borrelia All-In-One ViraChip Test Kit

B. Predicate 510(k) Number(s):

K220016

C. Comparison with Predicate(s):

Item	Device:	Predicate:
	iDart Lyme IgG ImmunoBlot Kit	Viramed Borrelia All-In-One ViraChip Test Kit
(K220016)
Similarities
Intended
Use	The iDartTM Lyme IgG ImmunoBlot Kit is an
immunoblot assay intended for the in vitro
qualitative detection of IgG antibodies to Borrelia
burgdorferi in human serum. The iDart Lyme IgG
ImmunoBlot Kit is intended to detect antibodies to
LSA and multiple other B. burgdorferi antigens
following a modified two-tier test methodology.
Positive results from the iDart Lyme IgG
ImmunoBlot Kit are supportive evidence for the
presence of antibodies and exposure to B.
burgdorferi. Negative results do not preclude
infection with B. burgdorferi. iDartTM Lyme IgG
ImmunoBlot Kit is intended to aid in the diagnosis
of Lyme disease and the test kit should only be
used on samples from patients with clinical
history, signs and symptoms consistent with Lyme
disease. The iDart Lyme IgG Immunoblot Kit is not
intended as a screen for asymptomatic patients.
Test results are to be used in conjunction with
information obtained from the patient's clinical
evaluation and other diagnostic procedures.
Test results are to be used in conjunction with
information obtained from the patient's clinical
evaluation and other diagnostic procedures.
For in vitro diagnostic use only
For professional use only
For prescription use only	The Viramed Biotech AG Borrelia All-In-One ViraChip
is an in vitro qualitative microarray
assay for the detection of IgM and IgG antibodies to
Borrelia burgdorferi in human serum. The assay is
intended for testing serum samples from symptomatic
patients or those suspected of Lyme Disease. It is
intended to detect antibodies to VIsE and multiple
other B. burgdorferi antigens following a modified two-
tier test methodology. Positive results from the
Viramed Biotech AG Borrelia All-In-One ViraChip are
supportive evidence for the presence of antibodies
and exposure to B. burgdorferi, the causative agent
for Lyme disease. Negative results do not preclude
infection with B. burgdorferi. Test results are to be
used in conjunction with information obtained from the
patient's clinical evaluation and other diagnostic
procedures as an aid in diagnosis of Lyme disease.
The Viramed Biotech AG Borrelia All-In-One ViraChip
Test must be used with a ViraChip
Reader and the ViraChip Software.
Specimen
Type	Serum	Serum
Antibodies
Detected	IgG	IgM and IgG
Controls	Positive Control serum, Negative Control Serum	Positive Control serum, Negative Control Serum
Method	Qualitative	Qualitative

6

Differences
ltem	Device	Predicate
Assay
Technology	ImmunoAssay	Antigen Coated wells
(Microarrays)
Antigens	Recombinant antigenic proteins -
LSA antigen (chimeric VIsE peptide termed the
Lyme Screen Antigen), 93 kD, 66 kD, 58 kD, 45
kD, 41 kD, 39 kD, 34 kD, 31 kD, 30 kD, 28 kD, 23
kD and 18 kD.	VlsE, 93 kD, 58 kD, 45
kD, 39 kD, 30 kD, 23
kD, 21 kD, 19 kD, 18
kD, and 17 kD antigens of B.burgdorferi
Procedure	Line Blot Assay Borrelia
IgG antibodies to specific antigen bands.
Wash between sample
and conjugate incubation steps,
incubate with substrate	Wash after Sample and
Conjugate Step
Sample
Volume	10 µl neat serum in 1000 ml sample diluent	Samples diluted 1:76 and 100 µL added per well
Reagents	Sample diluent, Wash Buffer, Milk powder,
Conjugate Buffer, Substrate solution	10X Wash Buffer, Sample Buffer,
Chromogen/Substrate Solution
Result
Generation	Manual reading	Automated with ViraChip Reader

STANDARDS/GUIDANCE DOCUMENTS REFERENCED: VI.

Establishing the Performance Characteristics of In Vitro Diagnostic Devices for the Detection of Antibodies to Borrelia burgdorferi - Guidance for Industry and FDA Staff - MARCH 2013

PERFORMANCE CHARACTERISTICS (IF/WHEN APPLICABLE): VII.

A. Analytical Performance:

1. Reproducibility:

The iDart™ Lyme IgG ImmunoBlot Kit was tested in a blind study to evaluate reproducibility across 3 separate sites each with 2 operators over 5 days, 2 runs a day, using a panel of blinded and coded samples of negative, moderate negative, high negative, moderate positive and high positive samples for IgG. It is concluded that there was 100% agreement of all bands among all runs, all days and across 3 sites for the iDart™ Lyme IgG ImmunoBlot Kit (See Table 1).

| Sample # | Sample Type | Number of
Bands
Present/sample | lgG | # of Samples
(+) | Expected
Result | % samples
matched to
expected result |
|----------|-------------------|--------------------------------------|-----|---------------------|--------------------|--------------------------------------------|
| GA | High Positive | 13 | P | 90/90 | P | 100% |
| GB | Moderate Positive | 10 | P | 90/90 | P | 100% |
| GC | Negative-1 | 5 | N | 0/90 | N | 100% |
| GD | Negative-2 | 0 | N | 0/90 | N | 100% |
| GE | Negative-3 | 1 | N | 0/90 | N | 100% |
| GF | Low Positive | 5 | P | 90/90 | p | 100% |

Table 1. Reproducibility Study Summary - Overall - All Sites - 6 operators/5 days/ 3 replicates

7

2. Analytical Specificity/Interference:

Analytical Specificity

Table 2 and 3 below shows the results of testing iDart™ Lyme IgG ImmunoBlot Kit on samples collected from a population of 313 apparently healthy individuals from endemic areas, and 112 samples collected from healthy individuals in non-endemic areas in the US.

Source	N (313)	IgG Positive
CDC	62	0
BAY AREA LYME FOUNDATION
(NY, MA, WI)	251	2
TOTAL	313	2
Specificity		99.36%

Table 2: iDart Lyme IgG ImmunoBlot results for samples collected from healthy individuals in endemic areas.

Table 3: iDart Lyme IgG ImmunoBlot results for samples collected from healthy individuals in non-endemic areas.

Source	N (112)	IgG Positive
CDC	62	0
CA	50	0
TOTAL	112	0
Specificity		100%

Cross-Reactivity Study

A cross reactivity study was performed on specimens known to contain potentially cross-reactive antibodies to Lyme infection. Serum samples from patients with bacterial/viral infections and sera from patients with diagnoses that can be confused with the late manifestations of Lyme disease were tested. Based on the data presented in Table 4, there was no cross-reactivity with antibodies to all non- Borrelia pathogens or autoimmune diseases tested with the iDart™ Lyme IgG ImmunoBlot Kit.

Table 4: iDart™ Lyme IgG ImmunoBlot Kit - Cross Reactivity
	Disease State		IgG			% Cross-
Source		N (376)	LSA	2+
Bands	IgG
Positive	reactivity
	Fibromyalgia	15	0	0	0	0%
	Mononucleosis	15	0	1	0	0%
CDC	Multiple sclerosis	15	0	0	0	0%
	Rheumatoid arthritis	15	0	0	0	0%
	Severe periodontitis	15	0	0	0	0%
	Syphilis	15	0	0	0	0%
	Babesiosis	28	0	0	0	0%
	Bartonellosis	48	0	0	0	0%
IGeneX (CA)	Ehrlichiosis	5	0	0	0	0%
	Anaplasmosis	7	0	0	0	0%
	Rickettsiosis	22	0	0	0	0%
	Tick Borne Relapsing Fever	14	0	0	0	0%
	HIV*	12	0	0	0	0%
	RPR	23	0	2	0	0%
New York Biological	HSV1	8	0	1	0	0%
(NY)	HSV2	2	0	0	0	0%
	CMV	13	0	0	0	0%
	EBV	27	0	0	0	0%
	RSV	4	0	0	0	0%
BEI	FLU	21	0	0	0	0%
Kamineni Life Sciences Pvt.	Pregnant women	12	0	0	0	0%

8

Ltd, Hydrabad (India)	H. pylori	10	0	0	0	0%
Warde Medical Laboratory
(MI)	Parvovirus-19	10	0	0	0	0%
CDC	Leptospira	10	0	0	0	0%
False Positive			0	5	0	0%
Specificity			100%	98.67%	100%

Interference from Endogenous Analytes

The potential interfering effect of endogenous substances in patient samples when using the iDart Lyme IgG ImmunoBlot was evaluated using one positive, one low positive and one negative Borrelia IgG samples were spiked with the endogenous substances at the final concentrations listed in the table below. All samples were tested in singlicate. No interference was observed in he tested samples.

Table 5. Effect of Interference Substances on iDart™ Lyme IgG ImmunoBlot Kit

| Agent | Concentration in
serum | iDart™ Lyme IgG result | | | Effect on ImmunoBlot
Kit |
|---------------|---------------------------|------------------------|--------------|----------|-----------------------------|
| | | High Positive | Low Positive | Negative | |
| Bilirubin | 1mg/dL (low) | Positive | Positive | Negative | No effect |
| Bilirubin | 15mg/dL (high) | Positive | Positive | Negative | No effect |
| Albumin | 3.5g/dL (low) | Positive | Positive | Negative | No effect |
| Albumin | 5g/dL (high) | Positive | Positive | Negative | No effect |
| Cholesterol | 150mg/dL (low) | Positive | Positive | Negative | No effect |
| Cholesterol | 250mg/dL (high) | Positive | Positive | Negative | No effect |
| Triglycerides | 150mg/dL (low) | Positive | Positive | Negative | No effect |
| Triglycerides | 500mg/dL (high) | Positive | Positive | Negative | No effect |
| Hemoglobin | 10g/dL (low) | Positive | Positive | Negative | No effect |
| Hemoglobin | 20g/dL (low) | Positive | Positive | Negative | No effect |

1. Assay Reportable Range: Not applicable.
1. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods): Not applicable.
1. Detection Limit: Not applicable.
1. Assay Cut-Off: Not applicable.

B. Clinical Studies:

1. Method Comparison with comparators (STTT):
  The performance of the iDart™ Lyme lgG ImmunoBlot Kit for detection of Borrelial-specific antibodies was compared to FDA-cleared ElA and immunoblot as part of the standard two-tier test methodology (STTT) .Results are summarized below. A total of 768 serum samples were procured from two vendors and tested at three clinical sites. Table 6 bellow summarizes the distribution of samples per testing site and cohort.

9

Table 6: Sample distribution by clinical site and cohort.

| | Number of
Samples | Sample Type | Vendors Providing
Samples |
|--------|----------------------|-----------------------------------|------------------------------|
| Site 1 | 290 | Clinical serum samples | Bay Area Lyme Foundation |
| Site 2 | 37 | Clinical serum samples - Cohort 2 | IGeneX Inc. |
| Site 2 | 230 | Clinical serum samples - Cohort 3 | IGeneX Inc. |
| Site 3 | 211 | Clinical serum samples - Cohort 2 | IGeneX Inc. |

All samples were blinded, re-coded, and tested at the respective clinical sites as per the instructions for use for the iDart Lyme IgG ImmunoBlot Kit. Performance by cohort is summarized in tables 7 through 9.

Table 7: Performance Summary on prospective banked samples from Bay Area Lyme Foundation (n=290). iDart Lyme IgG ImmunoBlot test versus STTT.

		STTT
		Positive (+)	Negative (-)
iDart Lyme IgG ImmunoBlot	Positive (+)	19	36
	Negative (-)	1	234
	Total	20	270
	PPA (95% CI)	95.00% (76.39% – 99.11%)
	NPA (95% CI)	86.67% (82.09% – 90.21%)

Table 8: Performance Summary on samples from IGeneX Inc. Cohort 2 (n=248). iDart Lyme IgG ImmunoBlot test versus STTT.

		STTT
		Positive (+)	Negative (-)
iDart Lyme IgG ImmunoBlot	Positive (+)	114	12
	Negative (-)	6	116
	Total	120	128
	PPA (95% CI)	95.00% (89.52% – 97.69%)
	NPA (95% CI)	90.63% (84.33% - 94.56%)

Table 9: Performance Summary on samples from IGeneX Inc. Cohort 3 (n=230). iDart Lyme IgG ImmunoBlot test versus STTT.

		STTT
		Positive (+)	Negative (-)
iDart Lyme IgG ImmunoBlot	Positive (+)	10	7
	Negative (-)	1	212
	Total	11	219
	PPA (95% CI)	90.91% (62.27% – 98.38%)
	NPA (95% CI)	96.80% (93.55% – 98.44%)

2. Clinical Sensitivity/Specificity:

CDC Serum Panel:

A Panel of 280 serum samples was received from CDC. These samples were from patients diagnosed with Lyme Disease at different stages (Stages 1, 2, and 3), Lyme disease look-like infectious mononucleosis, multiple sclerosis, rheumatoid arthritis, fibromyalgia and severe periodontitis), and from healthy controls living in endemic and non-endemic regions of Lyme disease. Results are analyzed according to disease stages and compared to STTT (See Table 10).

10

| Disease
Stage | Stage I | | Stage II | | Stage III | | Overall | | Healthy
controls | | Disease
Controls | |
|------------------|---------|--------|----------|--------|-----------|------|---------|--------|---------------------|------|---------------------|------|
| N | 60 | | 10 | | 20 | | 90 | | 100 | | 90 | |
| Test Kit | iDart | STTT | iDart | STTT | iDart | STTT | iDart | STTT | iDart | STTT | iDart | STTT |
| Positive | 35 | 18 | 9 | 9 | 20 | 20 | 64 | 47 | 0 | 0 | 0 | 0 |
| Negative | 25 | 42 | 1 | 1 | 0 | 0 | 26 | 43 | 100 | 100 | 90 | 90 |
| Sensitivity | 58.33% | 30.00% | 90.00% | 90.00% | 100% | 100% | 71.11% | 52.22% | | | | |
| Specificity | | | | | | | | | 100% | 100% | 100% | 100% |

Table 10. iDart Lyme IgG ImmunoBlot Kit performance using the CDC Reference Panel

3. Other Clinical Supportive Data:

Fresh and Frozen Samples Comparison Study:

72 decoded left-over patient serum samples were tested twice with iDart Lyme lgG ImmunmoBlot Test Kit, pre and post freezing. As shown below in Table 11, after freezing, all IgG positive samples remained positive and all negative samples remained negative. Clinical Performance of iDart™ Lyme IgG ImmunoBlot Kit for fresh and frozen samples are comparable and the test kit can be used on both fresh and frozen samples.

Table 11. : Fresh and Frozen Samples Tested with iDart™Lyme IgG ImmunoBlot Kit

Sample Type	Tested	N	IgG
			LSA (+)	≥2 bands	IgG (+)	Neg
Fresh Samples	Within 2 weeks of collection
(stored refrigerated)	72	39	47	33	39
Frozen Samples	After being Frozen (9-44 days)	72	39	47	33	39

Antibody Class Specificity:

A study was conducted to demonstrate the specificity of the goat anti-human IgG Conjugate used in the iDart™ Lyme IgG ImmunoBlot Kit. 10 previously tested patient samples that includes 6 negatives and 4 positives were included study. 3 sets of anti-human lgG conjugate were prepared and tested:

1ug/ml of human IgM
. 1ug/ml of human IgG
Control no additives .

The data in Table 12 demonstrates the IgG antibody class specificity performance of iDart™ Lyme IgG ImmunoBlot Kit. All positive samples tested with goat anti-human IgG conjugate without treatment with human IgG or with treatment with human IgM remained positive; and all negative samples remained neqative.

11

| Sample

| None | | | Human IgM | | | Human IgG | | |

|-------------|------|---------------|--------|-----------|---------------|--------|-----------|---------------|--------|
| | LSA | Band
Group | Result | LSA | Band
Group | Result | LSA | Band
Group | Result |
| 1 | P | P | P | P | P | P | N | N | N |
| 2 | N | P | N | N | P | N | N | N | N |
| 3 | P | P | P | P | P | P | N | N | N |
| 4 | P | P | P | P | P | P | N | N | N |
| 5 | P | P | P | P | P | P | N | N | N |
| 6 | N | N | N | N | N | N | N | N | N |
| 7 | N | P | N | N | P | N | N | N | N |
| 8 | N | P | N | N | P | N | N | N | N |
| 9 | N | N | N | N | N | N | N | N | N |
| 10 | N | P | N | N | P | N | N | N | N |

Table 12. iDart Lyme IgG ImmunoBlot Antibody Class Specificity Study results

C. Clinical Cut-Off:

Not applicable.

D. Expected Values/Reference Range:

Not applicable.

PROPOSED LABELING: VIII.

The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10

CONCLUSION: IX.

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