K Number

Device Name

QUALICODE B. BURGDORFERI IGM WESTERN BLOT KIT, MODEL DK-C062-024

Manufacturer

IMMUNETICS, INC.

Date Cleared

1999-09-21

(174 days)

Product Code

LSR

Regulation Number

866.3830

Intended Use

The QualiCode™ B. burgdorferi IgM Western Blot Kit is an in vitro qualitative assay for the detection of human IgM antibodies reactive with Borrelia burgdorferi antigens present on a membrane strip. The QualiCode™ B. burgdorferi IgM Western Blot Kit is intended for supplemental testing of human serum specimens which yield positive or equivocal results on B. burgdorferi ELISA or IFA screening assays. QualiCode test results can provide additional, more specific evidence of infection with B. burgdorferi which may be useful in the diagnosis of Lyme disease. The QualiCode™ B. burgdorferi IgM Western Blot Kit can be used to test human sera for IgM antibodies during the acute phase of disease, within one month of infection, in accordance with CDC/ASTPHLD guidelines. During this period, patients may not yet have developed a detectable IgG response. Beyond one month from infection, an IgG Western Blot test is recommended. Patients should not be tested solely for IgM antibodies at this stage due to the higher probability of false-positive results. Symptomatic patients showing IgM but not IgG antibodies at an early stage in infection should be re-tested for IgG antibodies 2-4 weeks later.

Device Description

The device is a Western Blot assay. Proteins and other antigenic components of the Borrelia spirochete are fractionated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. The separated proteins are electrophoretically transferred from the gel to nitrocellulose membranes. which are subsequently blocked to minimize non-specific binding and cut into strips. These nitrocellulose strips with resolved Borrelia burgdorferi antigens are then reacted with diluted serum and controls (positive and negative sera of defined reactivity). During the incubation period, human antibodies specific to the B, burgdorferi antigens, if present in the sample or control, will bind to the antigen to which they have affinity. Unbound serum and non-specific antibodies are washed from the strip. Detection of bound IgM antibodies is accomplished by reacting and incubating the strips with a solution containing anti-human IgM antibodies conjugated with alkaline phosphatase. Unbound conjugate antibodies are removed by washing. The qualitative assessment of the detected IgM antibodies is then accomplished by the reaction of the alkaline phosphatase with a chemical substrate, which is cleaved into a colored, insoluble product that can be visualized. The determination of the reactivity of each unknown specimen is accomplished by comparison of the identified, visualized bands to the Band Identifying and Band Intensity Controls.

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

Not Found

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The device description and performance studies focus on a traditional Western Blot assay and its manual interpretation, with no mention of AI or ML.

Therapeutic

No.
The device is an in vitro diagnostic assay used for qualitative detection of antibodies, providing evidence for the diagnosis of Lyme disease; it does not treat or alleviate the disease.

Diagnostic

Yes
The device is described as an "in vitro qualitative assay for the detection of human IgM antibodies reactive with Borrelia burgdorferi antigens" and is intended for "supplemental testing" to "provide additional, more specific evidence of infection with B. burgdorferi which may be useful in the diagnosis of Lyme disease." This directly indicates its use in diagnosing a medical condition.

SaMD (Software as a Medical Device)

The device description clearly outlines a laboratory assay kit involving physical components like membrane strips, reagents, and a chemical reaction to visualize results. This is a hardware-based in vitro diagnostic device, not a software-only device.

IVD (In Vitro Diagnostic)

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

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the device is an "in vitro qualitative assay for the detection of human IgM antibodies reactive with Borrelia burgdorferi antigens present on a membrane strip." It also specifies that it is intended for "supplemental testing of human serum specimens." This clearly indicates that the device is used to examine specimens taken from the human body in vitro (outside the body) to provide information for diagnostic purposes (diagnosis of Lyme disease).
Device Description: The "Device Description" details a laboratory-based assay process involving the reaction of human serum with antigens on a membrane strip, followed by detection of bound antibodies using conjugated antibodies and a substrate. This is a typical description of an in vitro diagnostic test.
Performance Studies: The inclusion of performance studies evaluating specificity and sensitivity using human serum samples from different patient populations (normal, cross-reactive, Lyme disease) further confirms its use as a diagnostic tool for analyzing human specimens.

Therefore, based on the provided information, the QualiCode™ B. burgdorferi IgM Western Blot Kit fits the definition of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The QualiCode™ B. burgdorferi IgM Western Blot Kit can be used to test human sera for IgM antibodies during the acute phase of disease, within one month of infection, in accordance with CDC/ASTPHLD guidelines. During this period, patients may not yet have developed a detectable IgG response. Beyond one month from infection, an IgG Western Blot test is recommended. Patients should not be tested solely for IgM antibodies at this stage due to the higher probability of false-positive results. Symptomatic patients showing IgM but not IgG antibodies at an early stage in infection should be re-tested for IgG antibodies 2-4 weeks later.

The Immunetics QualiCode" B. burgdorferi IgM Western Blot Kit is intended for use in testing human serum samples which have demonstrated positive or eguivocal responses using EIA or IFA test procedures to provide supportive evidence of infection with Borrelia burgdorferi.

The QualiCode B. burgdorferi IgM Western Blot Kit can be used to test human sera for IqM antibodies during the acute phase of disease, within one month of infection, in accordance with CDC/ASTPHLD During this period, patients may not yet have developed quidelines. a detectable IgG response. Beyond one month from infection, an IgG Western Blot test is recommended. Patients should not be tested solely for IgM antibodies at this stage due to the higher probability of false positive results. Symptomatic patients showing IgM but not IqG antibodies at an early stage in infection should be re-tested for IgG antibodies 2-4 weeks later.

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

LSR

Device Description

During the incubation period, human antibodies specific to the B, burgdorferi antigens, if present in the sample or control, will bind to the antigen to which they have affinity. Unbound serum and non-specific antibodies are washed from the strip. Detection of bound IgM antibodies is accomplished by reacting and incubating the strips with a solution containing anti-human IgM antibodies conjugated with alkaline phosphatase. Unbound conjugate antibodies are removed by washing. The qualitative assessment of the detected IgM antibodies is then accomplished by the reaction of the alkaline phosphatase with a chemical substrate, which is cleaved into a colored, insoluble product that can be visualized. The determination of the reactivity of each unknown specimen is accomplished by comparison of the identified, visualized bands to the Band Identifying and Band Intensity Controls.

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

Not Found

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

Not Found

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

Not Found

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

Expected values:
Three (3) investigational sites, including Immunetics and two (2) independent off-site investigators, assayed samples from the following patient populations:

Normal population (n=430) comprised of samples from Lyme disease endemic (n=279) and non-endemic (n=151) regions.
Cross Reactive Panel (n=172) comprised of samples from patients with diseases other than Lyme disease that may be cross - reactive.
Lyme Disease Panel (n=186) comprised of samples from patients with a clinical diagnosis of Lyme disease, based on presence of erythema migrans or one or more symptoms of late Lyme disease, and which tested positive or equivocal on ELISA or other screening assays for B. burgdorferi antibodies.

Specificity (study type: Performance against normal and cross-reactive panels):
Specificity of the device was determined from analysis of results of testing normal donor specimens from endemic and non-endemic regions and potentially cross reactive disease specimens (602 total samples). The specificity values derived from testing each population are shown in the table below in the "% Negative" column:

Sample Type	n	% Positive	% Negative	95% CI
Normal - endemic region	279	5%	95%	92-97 %
Normal - non-endemic region	151	4%	96%	92-98 %
Normal - overall	430	4%	96%	93-97 %
Cross - Reactive Panel	172	13%	87%	88-96%

Sensitivity (study type: Performance against well-characterized Lyme disease sera):
Sensitivity of the device was determined by testing a total of 186 well-characterized sera from patients with Lyme disease, which had been drawn at different times after onset of disease. Sensitivity for each time after onset category is shown in the table below in the "% Positive" column:

Draw Time (months)	n	% Positive	% Negative	95% CI
Unknown	54	74%	26%	60-85 %
12	17	47%	53%	25-72 %

Reproducibility (study type: Inter-lot, Inter-run, Inter-reader):

Inter-lot reproducibility: Assaying a panel of 20 specimens, including positive, weakly reactive and negative sera, on three lots of the Immunetics QualiCode™ kit. There was 80% agreement between interpretations from the three kit lots. The reproducibility of scoring of individual bands between the three lots varied from 80% to 100%, averaging 88%.
Inter-run reproducibility: Assaying the 20 specimen panel twice, on separate days, at each of three sites. There was 85% agreement overall between interpretations from the two runs averaged over all three sites. The reproducibility of individual band scoring between the two runs averaged over all three sites varied between 75% and 93%, with an average of 85% over all three bands.
Inter-reader reproducibility: Assaying the 20 specimen panel twice, on separate days, at each of three sites, with each strip interpreted by two indevendent readers. There was 93% agreement overall between interpretations from the two readers averaged over the three sites. Band scoring reproducibility varied between 97% and 98% averaged over the three sites, with an average of 97% over all three bands.

Study	n	% Interpretation Agreement
Inter-lot	20	80
Inter-run	20	85
Inter-reader	20	93

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

Specificity:
- Normal - endemic region: 95%
- Normal - non-endemic region: 96%
- Normal - overall: 96%
- Cross - Reactive Panel: 87%
Sensitivity (% Positive):
- Unknown draw time: 74%
- 12 months draw time: 47%
Reproducibility (% Interpretation Agreement):
- Inter-lot: 80%
- Inter-run: 85%
- Inter-reader: 93%

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

Not Found

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

Not Found

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

Not Found

Regulation Number and Section

§ 866.3830

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

510(k) Safety and Effectiveness Summary

Submitted by:

Immunetics, Inc. 63 Rogers Street Cambridge, MA 02142

Contact Person:

Andrew E. Levin, Ph.D. Scientific Director

(617) 492 - 5416

Date of Preparation:

September 9, 1999

1. Name and Address of Owner/Operator and Manufacturer

Immunetics, Inc. 63 Rogers Street Cambridge, MA 02142

2. Product Name

Trade Name: QualiCode™ B. burgdorferi IgM Western Blot Kit

Common Name: B. burgdorferi IgM Western Blot Kit

3. Claim of Substantial Equivalence

The characterized samples used for the establishment of Substantial Equivalence were obtained from patients with a clinical diagnosis of Lyme Disease in accordance with the CDC case definition, i.e. based on the presence of EM (erythema migrans) or the presentation of late Lyme clinical manifestations (e.g., arthritic, cardiac, or neurological symptoms). Infection was confirmed by culture of Borrelia from biopsies in many examples.

Each of the clinical trial sites provided specimens that were well characterized by the site using Lyme-specific serological analyses, including EIA and/or Western Blot testing. In particular, specimens selected for the trial were required to have tested positive or indeterminate on a B. burgdorferi screening assay, typically an ELISA, in accordance with the two-tier testing protocol recommended by CDC/ASTPHLD (CDC (1995) "Recommendations for test performance and interpretation from the Second

National Conference on Serologic Diagnosis of Lyme Disease", Morbid. Mort. Weekly Rep. 44:590-591.).

Substantial equivalence of this device is based on the assessment of performance of the device in these clinical trials in which the well-characterized, archived Lyme Disease specimens, the Centers for Disease Control Lyme Disease Serum Panel, normal donor specimens (from endemic and non-endemic regions), and samples from diverse disease conditions were analyzed.

4. Description

5. Intended use

antibodies at this stage due to the higher probability of false-positive results. Symptomatic patients showing IgM but not IgG antibodies at an early stage in infection should be re-tested for IgG antibodies 2-4 weeks later.

Summary of Performance 6.

From a summary of the clinical trial data, the following performance characteristics are described:

Expected values

Three (3) investigational sites, including Immunetics and two (2) independent off-site investigators, assayed samples from the following patient populations:

1. Normal population (n=430) comprised of samples from Lyme disease endemic (n=279) and non-endemic (n=151) regions.
1. Cross Reactive Panel (n=172) comprised of samples from patients with diseases other than Lyme disease that may be cross - reactive.
1. Lyme Disease Panel (n=186) comprised of samples from patients with a clinical diagnosis of Lyme disease, based on presence of erythema migrans or one or more symptoms of late Lyme disease, and which tested positive or equivocal on ELISA or other screening assays for B. burgdorferi antibodies.

Specificity

Specificity of the device was determined from analysis of results of testing normal donor specimens from endemic and non-endemic regions and potentially cross reactive disease specimens (602 total samples). The specificity values derived from testing each population are shown in the table below in the "% Negative" column:

Sample Type	n	% Positive	% Negative	95% CI
Normal - endemic region	279	5%	95%	92-97 %
Normal - non-endemic region	151	4%	96%	92-98 %
Normal - overall	430	4%	96%	93-97 %
Cross - Reactive Panel	172	13%	87%	88-96%

Sensitivity

Sensitivity of the device was determined by testing a total of 186 well-characterized sera from patients with Lyme disease, which had been drawn at different times after onset of disease. Sensitivity for each time after onset category is shown in the table below in the "% Positive" column:

Sensitivity of the Immunetics, Inc. QualiCode™ B. burgdorferi IgM Western Blot Kit vs. Time after Onset

Draw Time (months)	n	% Positive	% Negative	95% CI
Unknown	54	74%	26%	60-85 %
12	17	47%	53%	25-72 %

Reproducibility

Inter-lot reproducibility was determined by assaying a panel of 20 specimens, including positive, weakly reactive and negative sera, on three lots of the Immunetics QualiCode™ kit. There was 80% agreement between interpretations from the three kit lots. The reproducibility of scoring of individual bands between the three lots varied from 80% to 100%, averaging 88%.

Inter-run reproducibility was determined by assaying the 20 specimen panel twice, on separate days, at each of three sites. There was 85% agreement overall between interpretations from the two runs averaged over all three sites. The reproducibility of individual band scoring between the two runs averaged over all three sites varied between 75% and 93%, with an average of 85% over all three bands.

Inter-reader reproducibility was determined by assaying the 20 specimen panel twice, on separate days, at each of three sites, with each strip interpreted by two indevendent readers. There was 93% agreement overall between interpretations from the two readers averaged over the three sites. Band scoring reproducibility varied between 97% and 98% averaged over the three sites, with an average of 97% over all three bands.

Study	n	% Interpretation Agreement
Inter-lot	20	80
Inter-run	20	85
Inter-reader	20	93

7. Conclusions

Based on the clinical performance, this device has been shown to be safe and effective for the intended use in the qualitative detection of human Immunoglobulin M (IgM) antibodies in serum or plasma to Borrelia burgdorferi antigens, and as a supplemental, more specific, test to aid in the diagnosis of infection or exposure to Borrelia burgdorferi, the causative agent of Lyme disease.

Image /page/4/Picture/1 description: The image is a seal for the Department of Health & Human Services - USA. The seal is circular and contains the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. In the center of the seal is an abstract image of an eagle.

SEP 2 1 1999

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Andrew E. Levin. Ph.D. Scientific Director Immunetics, Inc. 63 Rogers Street Cambridge, Massachusetts 02142

Re: K991062

Trade Name: QualiCode™ B. burgdorferi IgM Western Blot Kit Regulatory Class: II Product Code: LSR Dated: July 8, 1999 Received: July 9, 1999

Dear Dr. Levin:

We have reviewed your Section 510(k) notification of intent to market the device referenced above and we 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). 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.

If your device is classified (see above) into either class II (Special Controls) or class III (Premarket Approval), it may be subject to such additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 895. A substantially equivalent determination assumes compliance with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (OS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic OS inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to comply with the GMP regulation may result in regulatory action. In addition. FDA may publish further announcements concerning your device in the Federal Register. Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal laws or regulations.

Page 2

Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device may require a CLIA complexity categorization. To determine if it does, you should contact the Centers for Disease Control and Prevention (CDC) at (770)488-7655.

This letter will allow you to begin marketing your device as described in your 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801 and additionally 809.10 for in vitro diagnostic devices), please contact the Office of Compliance at (301) 594-4588. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at (301) 594-4639. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll free number (800) 638-2041 or at (301) 443-6597 or at its internet address "http://www.fda.gov/cdrh/dsmamain.html"

Sincerely yours,

Steven Autman

Steven I. Gutman, M.D., M.B.A. Director Division of Clinical Laboratory Devices Office of Device Evaluation Center for Devices and Radiological Health

Enclosure

510(k) Number (if known):

Device Name: QualiCode B. burgdorferi IgM Western Blot Kit

Indications For Use:

(PLEASE DO NOT WRITE BELOW THIS LINE - CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of Device Evaluation (ODE)

Woody Dubres

Prescription Use
(Per 21 CFR 801.109)

Over-The-Counter Use

(Optional format 1-2-96)