← Product Code [LSR](/submissions/MI/subpart-d%E2%80%94serological-reagents/LSR) · K172722

# Anti-Borrelia burgdorferi US EUROLINE-WB (IgM) (K172722)

_Euroimmun Us, Inc. · LSR · Dec 10, 2017 · Microbiology · SESE_

**Canonical URL:** https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/LSR/K172722

## Device Facts

- **Applicant:** Euroimmun Us, Inc.
- **Product Code:** [LSR](/submissions/MI/subpart-d%E2%80%94serological-reagents/LSR.md)
- **Decision Date:** Dec 10, 2017
- **Decision:** SESE
- **Submission Type:** Traditional
- **Regulation:** 21 CFR 866.3830
- **Device Class:** Class 2
- **Review Panel:** Microbiology
- **Attributes:** Software as a Medical Device

## Intended Use

The Anti-Borrelia burgdorferi US EUROLINE-WB (IgM) kit is a Western blot assay intended for the qualitative determination of IgM class antibodies against Borrelia burgdorferi in human serum and plasma (K⁺-EDTA, Li⁺-heparin, Na⁺-citrate) samples that have been found positive or equivocal/borderline using an enzyme immunoassay (EIA) or immunofluorescence assay (IFA) test procedure for Borrelia burgdorferi antibodies. Results can be read manually or automated utilizing EUROLineScan. This test is used as an aid in the diagnosis of infections with Borrelia burgdorferi and the associated diseases, in conjunction with other laboratory and clinical findings.

## Device Story

Western blot assay for qualitative detection of IgM antibodies against Borrelia burgdorferi. Input: human serum or plasma samples (K-EDTA, Li-heparin, Na-citrate). Process: electrophoretically separated B. burgdorferi antigen extracts on nitrocellulose strips; patient sample incubation; binding of specific IgM to antigens; enzyme-labeled anti-human IgM conjugate incubation; color reaction with NBT/BCIP substrate. Output: visual presence of specific protein bands (p25, p39, p41). Used in clinical laboratories; operated by trained technicians. Results read manually or via EUROLineScan software. Aids clinicians in Lyme disease diagnosis by confirming EIA/IFA results; supports clinical decision-making regarding infection status.

## Clinical Evidence

Clinical sensitivity evaluated using 101 characterized Lyme disease specimens across various disease stages; overall sensitivity 44.6%. Prospective method comparison study (n=304) against predicate showed 96.9% positive agreement and 99.5% negative agreement. Analytical specificity tested in endemic (n=98) and non-endemic (n=100) populations; cross-reactivity panel (n=155) showed no interference from common pathogens or rheumatoid arthritis. CDC panel testing (n=34) provided additional performance characterization.

## Technological Characteristics

Western blot assay; qualitative detection of IgM antibodies; utilizes antigen-coated strips; manual or automated readout via EUROLineScan software.

## Regulatory 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), the Treponema pallidum immobilization test (T.P.I.), and other treponemal tests used to identify antibodies to Treponema pallidum directly from infecting treponemal organisms in serum. The identification aids in the diagnosis of syphilis caused by bacteria belonging to the genus Treponema and provides epidemiological information on syphilis.

## Submission Summary (Full Text)

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>
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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

A. 510(k) Number: K172722
B. Purpose for Submission: To obtain a substantial equivalence determination and FDA clearance for a new device
C. Measurand: Anti-Borrelia burgdorferi (IgM) antibodies
D. Type of Test: Enzyme Immunoassay
E. Applicant: EUROIMMUN US
F. Proprietary and Established Names: Anti-Borreliaburgdorferi(IgM) antibodies

## G. Regulatory Information:

1. Regulation section: 21 CFR 866.3830; Treponema pallidum treponemal test reagents
2. Classification: Class II
3. Product code: LSR; Reagent, Borrelia Serological Reagent
4. Panel: Microbiology

## H. Intended Use:

1. Intended use(s): The Anti-Borrelia burgdorferi US EUROLINE-WB (IgM) kit is a Western blot assay intended for the qualitative determination of IgM class antibodies against Borrelia burgdorferi in human serum and plasma (K⁺-EDTA, Li⁺-heparin, Na⁺-citrate) samples that have been found positive or equivocal/borderline using an enzyme immunoassay (EIA) or immunofluorescence assay (IFA) test procedure for Borrelia burgdorferi antibodies. Results can be read manually or automated utilizing EUROLineScan. This test is used as an aid in the diagnosis of infections with Borrelia burgdorferi and the associated diseases, in conjunction with other laboratory and clinical findings.
2. Indication(s) for use: Same as Intended Use
3. Special conditions for use statement(s): N/A
4. Special instrument requirements: N/A

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I. Device Description: The test kit contains test strips with electrophoretically separated antigen extracts of  $B$  burgdorferi. Each test strip contains an additional membrane chip coated with p41 flagellin antigen. The blot strips will be blocked and incubated in the first reaction step with diluted patient samples. In the case of positive samples, specific antibodies of the class IgM will bind to the antigens. To detect the bound antibodies, a second incubation is carried out using an enzyme-labeled anti-human IgM (enzyme conjugate), catalyzing a color reaction.

# J. Substantial Equivalence Information:

1. Predicate device name(s): MarDx Lyme Disease (IgM) Marblot
2. Predicate  $510(\mathrm{k})$  number(s): K951709
3. Comparison with predicate:

Table 1: Similarities and Differences with the Predicate

|  Similarities  |   |   |
| --- | --- | --- |
|  Item | Device | Predicate  |
|  Intended Use | Detection of IgM antibodies to Borrelia burgdorferi | Same  |
|  Assay format | Qualitative | Same  |
|  Technology/Procedure | Serum incubation with antigen coated strips followed by a wash step, incubation with an anti-human IgM enzyme-conjugate; wash step, incubation with substrate, wash step, air drying and visual reading. | Same  |
|  Antigens | Antigens of Borrelia Burgdorferi (strain B31) are separated in the presence of sodium dodecyl sulfate (SDS) by polyacrylamide gel electrophoresis. The resolved protein bands are then transferred to a nitrocellulose membrane. | Same  |
|  Conjugate | Alkaline phosphatase anti-human IgM | Same  |
|  Substrate | NBT/BCIP | Same  |
|  Differences  |   |   |
| --- | --- | --- |
|  Item | Device | Predicate  |
|  Sample | 30 μl Serum or Plasma 1:51 dilution | 20 μl Serum undiluted  |
|  Incubation times | 30 – 30 – 10 min | 30 – 15 – (4 - 12) min  |
|  Controls | Evaluation matrix with control strip (test strip incubated with a positive control serum) | Serum band locator Weakly reactive control Negative control  |

K. Standard/Guidance Document Referenced (if applicable): N/A

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L. Test Principle: Enzyme Immunoassay

M. Performance Characteristics (if/when applicable):

1. Analytical performance:

a. Precision/Reproducibility:

Precision/Repeatability: Repeatability was investigated by repeated determinations of seven characterized samples. The samples were tested on four different days with two runs per day, two replicates per run according to the package insert. Repeatability was found to be sufficient as no positive sample was found negative and vice versa.

Table 2: Precision Study

|  n = 16 | Anti-Borrelia burgdorferi US EUROLINE-WB (IgM)  |   |   |   |   |   |   |
| --- | --- | --- | --- | --- | --- | --- | --- |
|   |  Sample 1 | Sample 2 | Sample 3 | Sample 4 | Sample 5 | Sample 6 | Sample 7  |
|  Characterization | negative | negative | negative | negative | negative | positive | positive  |
|  % negative | 100.0% | 100.0% | 100.0% | 100.0% | 100.0% | 0.0% | 0.0%  |
|  % positive | 0.0% | 0.0% | 0.0% | 0.0% | 0.0% | 100.0% | 100.0%  |

Table 2 (continued): Detailed Precision Study Data

|  Band(s) | Anti-Borrelia burgdorferi US EUROLINE-WB (IgM)  |   |   |
| --- | --- | --- | --- |
|   |  p25 | p39 | p41  |
|  Sample 1 | 0.0% | 0.0% | 0.0%  |
|  Sample 2 | 0.0% | 0.0% | 100.0%  |
|  Sample 3 | 0.0% | 0.0% | 100.0%  |
|  Sample 4 | 0.0% | 0.0% | 100.0%  |
|  Sample 5 | 0.0% | 0.0% | 100.0%  |
|  Sample 6 | 100.0% | 100.0% | 100.0%  |
|  Sample 7 | 0.0% | 100.0% | 100.0%  |

Reproducibility: Reproducibility was investigated by repeated determinations of seven characterized samples. The samples were tested on four different days with two runs per day, two replicates per run at three different sites according to the package insert. Reproducibility was found to be sufficient as no positive sample was found negative and vice versa.

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Table 3: Reproducibility Study

|  n = 48 | Anti-Borrelia burgdorferi US EUROLINE-WB (IgM)  |   |   |   |   |   |   |
| --- | --- | --- | --- | --- | --- | --- | --- |
|   |  Sample 1 | Sample 2 | Sample 3 | Sample 4 | Sample 5 | Sample 6 | Sample 7  |
|  Characterization | negative | negative | negative | negative | negative | positive | positive  |
|  % negative | 100.0% | 100.0% | 100.0% | 100.0% | 100.0% | 0.0% | 0.0%  |
|  % positive | 0.0% | 0.0% | 0.0% | 0.0% | 0.0% | 100.0% | 100.0%  |

Table 3 continued: Detailed Reproducibility Data

|  Band(s) | Anti-Borrelia burgdorferi US EUROLINE-WB (IgM)  |   |   |
| --- | --- | --- | --- |
|   |  p25 | p39 | p41  |
|  Sample 1 | 0.0% | 0.0% | 0.0%  |
|  Sample 2 | 0.0% | 0.0% | 100.0%  |
|  Sample 3 | 0.0% | 0.0% | 100.0%  |
|  Sample 4 | 0.0% | 0.0% | 100.0%  |
|  Sample 5 | 0.0% | 0.0% | 100.0%  |
|  Sample 6 | 100.0% | 100.0% | 100.0%  |
|  Sample 7 | 0.0% | 100.0% | 100.0%  |

b. Linearity/assay reportable range: N/A
c. Traceability, Stability, Expected values (controls, calibrators, or methods): N/A
d. Detection limit: N/A
e. Analytical specificity:

Normal Population Study: Testing of samples from asymptomatic population was performed in endemic and non-endemic regions with the Anti-Borrelia burgdorferi US EUROLINE-WB (IgM). A panel of 98 samples from the endemic region (Pennsylvania; 89 men and 9 women with an average age of 34 years; age range: 18 – 56 years) and a panel of 100 samples from a non-endemic region (Tennessee; 82 men and 18 women with an average age of 38 years; age range: 3 – 62 years) were tested. The results are shown below.

Table 4: Analytical Specificity Study

|  Panel | n | Anti-Borrelia burgdorferi US EUROLINE-WB (IgM)  |   |
| --- | --- | --- | --- |
|   |   |  Positive (%) | Negative (%)  |
|  Endemic | 98 | 5 (5.1%) | 93 (94.9%)  |
|  Non-endemic | 100 | 4 (4.0%) | 96 (96.0%)  |

Cross-Reactivity: Cross-reactivity was investigated using characterized samples from the following groups as shown in the table below. All the samples tested were found negative with the Anti-Borrelia burgdorferi US EUROLINE-WB (IgM) according to the CDC criteria of results interpretation.

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Table 5: Cross-Reactivity Study

|  Panel |  | n | Anti-Borrelia burgdorferi US EUROLINE-WB (IgM)  |
| --- | --- | --- | --- |
|   |   |   |  Negative (%)  |
|  EBV |  | 15 | 15 (100.0%)  |
|  HSV |  | 14 | 14 (100.0%)  |
|  Influenza viruses |  | 15 | 15 (100.0%)  |
|  H. pylori |  | 11 | 11 (100.0%)  |
|  Measles |  | 15 | 15 (100.0%)  |
|  Parvovirus B19 |  | 12 | 12 (100.0%)  |
|  Rubella |  | 14 | 14 (100.0%)  |
|  Treponema |  | 10 | 10 (100.0%)  |
|  CMV |  | 10 | 10 (100.0%)  |
|  Babesiosis |  | 3 | 3 (100.0%)  |
|  Anaplasmosis (ehrlichiosis) |  | 10 | 10 (100.0%)  |
|  Rickettsial diseases |  | 4 | 4 (100.0%)  |
|  Rheumatoid arthritis |  | 22 | 22 (100.0%)  |
|  Total |  | 155 | 155 (100.0%)  |

Note: The results obtained with babesiosis (3) and ricketsial diseases (4) samples are not conclusive as enough samples were not tested.

Interferences: Hemolytic, lipemic and icteric samples showed no influence at the result up to a concentration of 500 mg/dl for hemoglobin, 2000 mg/dl for triglycerides and 40 mg/dl for bilirubin in this test system. Interferences with albumin, intralipids, and cholesterol have not been investigated.

f. Assay cut-off:

Band Cut-off Determination: The cut-off of the Anti-Borrelia burgdorferi US EUROLINE-WB (IgM) test system is defined as the lowest limit of a clearly visible band. Because visual examination by operator might be subjective, the reaction control card was developed to standardize strip evaluation. Alternatively the EUROLineScan software was established to allow for automated evaluation.

2. Comparison studies:

a. Method comparison with predicate device:

Prospective Study: A prospective study was performed at 3 different sites in the U.S. Samples were tested as per the CDC two-tiered testing for Lyme disease algorithm. All positive and equivocal samples by the first-tier EIA were tested with the Anti-Borrelia burgdorferi US EUROLINE-WB (IgM) and the predicate IgM Western blot. Results are interpreted by the CDC criteria.

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Table 6: Percent Agreement with Predicate Device

|  n = 304 | Predicate IgM WB  |   |   |
| --- | --- | --- | --- |
|   |   |  Positive | Negative  |
|  Anti-Borrelia burgdorferi US EUROLINE-WB (IgM) | Positive | 95 | 1  |
|   |  Negative | 3 | 205  |

Positive agreement 95/98 = 96.9% 95% C.I.: 91.3% - 99.4%
Negative agreement 205/206 = 99.5% 95% C.I.: 97.3% - 100.0%

b. Matrix Comparison

Serum/Plasma Comparison: The use of K⁺-EDTA, Li⁺-heparin, Na⁺-citrate plasma samples was confirmed by a correlation of 10 sample sets of serum and corresponding plasma. The sample sets were selected to cover both negative and positive results. The results of the plasma samples and the corresponding serum sample were compared and found to be sufficient as no positive sample was found negative and vice versa.

3. Clinical studies:

a. Clinical Sensitivity:

Clinical/Diagnostic Sensitivity Study: A study, consisting of 101 clinically characterized Lyme disease specimens, was conducted at the manufacturer's site with the Anti-Borrelia burgdorferi US EUROLINE-WB (IgM) test device. These specimens contain samples from early, early disseminated and late phases of the disease. The panel consisted of 35 men, 58 women and 8 unknowns. The age ranged from 16 - 87 years with a mean age of 45 years.

Table 7: Case Confirmed Lyme Disease Samples

|  Interval | n | Anti-Borrelia burgdorferi US EUROLINE-WB (IgM) |   |   | Predicate IgM WB  |   |   |
| --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |  Positive | % | 95% C.I. | Positive | % | 95% C.I.  |
|  <1 month | 11 | 7 | 63.6% | 30.8 - 89.1% | 4 | 36.4% | 10.9 - 69.2%  |
|  >1-3 months | 23 | 12 | 52.2% | 30.6 - 73.2% | 9 | 39.1% | 19.7 - 61.5%  |
|  >3-12 months | 38 | 15 | 39.5% | 24.0 - 56.6% | 8 | 21.1% | 9.6 - 37.3%  |
|  >12 months | 29 | 11 | 37.9% | 20.7 - 57.7% | 4 | 13.8% | 3.9 - 31.7%  |
|  Overall | 101 | 45 | 44.6% | 34.7 - 54.8% | 25 | 24.8% | 16.7 - 34.3%  |

CDC Panel Results: Thirty-four (34) sera of patients with clinically characterized borreliosis in different disease stages and five normals, obtained from the Centers for Disease Control and Prevention, Atlanta, GA, USA, were tested with the Anti-Borrelia burgdorferi US EUROLINE-WB (IgM) in parallel with the predicate device. The results stratified by time after onset of infection are shown below.

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Table 8: Testing of CDC Lyme Disease Panel

|  Interval | n | Anti-Borrelia burgdorferi US EUROLINE-WB (IgM) |   |   | Predicate IgM WB  |   |   |
| --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |  Negative | % | 95% C.I. | Negative | % | 95% C.I.  |
|  Normals | 5 | 5 | 100.0% | 47.8 – 100.0% | 5 | 100.0% | 47.8 – 100.0%  |

Table 8 continued: Detailed Analysis of CDC Lyme Disease Panel Testing

|  Interval | n | Anti-Borrelia burgdorferi US EUROLINE-WB (IgM) |   |   | Predicate IgM WB  |   |   |
| --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |  Positive | % | 95% C.I. | Positive | % | 95% C.I.  |
|  <1 month | 6 | 3 | 50.0% | 11.8 - 88.2% | 3 | 50.0% | 11.8 - 88.2%  |
|  >1-3 months | 10 | 7 | 70.0% | 34.8 - 93.3% | 7 | 70.0% | 34.8 - 93.3%  |
|  >3-12 months | 12 | 2 | 16.7% | 2.1 - 48.4% | 2 | 16.7% | 2.1 - 48.4%  |
|  >12 months | 6 | 0 | 0.0% | 0.0 - 45.9% | 0 | 0.0% | 0.0 - 45.9%  |
|  Overall | 34 | 12 | 35.3% | 19.7 - 53.5% | 12 | 35.3% | 19.7 – 53.5%  |

Note: The results are presented as a means to convey further information on the performance of this assay with a masked, characterized serum panel. This does not imply an endorsement of the assay by the CDC.

b. Clinical specificity: N/A
c. Other clinical supportive data (when a. and b. are not applicable): N/A

4. Clinical cut-off: N/A
5. Expected values/Reference range:

Expected Values: The incidence of IgM antibodies and positivity in different populations as tested by the Anti-Borrelia burgdorferi US EUROLINE-WB (IgM) are presented below with available patient demographics.

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Table 9: Expected Values

|  Population |   | n | Gender | Mean Age Years (Range) | p25 | p39 | p41 | No. Pos. (%)  |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
|  Sensitivity Study | <1 mth | 11 | 4 men;7 women; | 45 (16-70) 1 unknown | 8 | 0 | 8 | 7 (63.6)  |
|   |  >1-3 mths | 23 | 4 men;18 women;4 unknown | 47 (21-80) 5 unknown | 15 | 4 | 20 | 12 (52.2)  |
|   |  >3-12 mths | 38 | 17 men;17 women;2 unknown | 46 (18-87) 3 unknown | 12 | 1 | 18 | 15 (39.5)  |
|   |  >yr(s) | 29 | 10 men;16 women;2 unknown | 41 (18-63) 2 unknown | 9 | 2 | 15 | 11 (37.9)  |
|   |  Total | 101 | 35 men;58 women;8 unknown | 45 (16-87) 11 unknown | 44 | 7 | 61 | 45 (44.6)  |
|  Prospective Study | Site 1 | 142 | 88 men;52 women;2 unknown | 51 (4-88) 4 unknown | 45 | 38 | 112 | 56 (39.4)  |
|   |  Site 2 | 97 | 51 men;46 women | 57 (20 - 75) | 33 | 5 | 53 | 29 (29.9)  |
|   |  Site 3 | 65 | 30 men;35 women | 60 (19-86) | 13 | 4 | 26 | 11 (16.9)  |
|   |  Total | 304 | 169 men;133 women;2 unknown | 56 (4-88) 4 unknown | 91 | 47 | 191 | 96 (31.6)  |
|  Normal Population Study | Endemic | 98 | 89 men;9 women | 34 (18-56) | 3 | 4 | 84 | 5 (5.1)  |
|   |  Non-Endemic | 100 | 82 men;18 women | 38 (3-62) | 2 | 2 | 87 | 4 (4.0)  |
|   |  Total | 198 | 171 men;27 women | 36 (3-62) | 5 | 6 | 171 | 9 (4.5)  |

Note: It is recommended that each laboratory determine its own normal range based on the population and equipment used.

# N. Proposed Labeling:

The labeling is sufficient and it satisfies the requirements of 21 CFR Parts 801 and 809, as applicable.

# O. Conclusion:

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

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**Source:** [https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/LSR/K172722](https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/LSR/K172722)

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