← Product Code [LSR](/submissions/MI/subpart-d%E2%80%94serological-reagents/LSR) · K142038

# EUROIMMUN LYME ELISA(IgG/IgM) (K142038)

_Euroimmun US · LSR · May 4, 2015 · Microbiology · SESE_

**Canonical URL:** https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/LSR/K142038

## Device Facts

- **Applicant:** Euroimmun US
- **Product Code:** [LSR](/submissions/MI/subpart-d%E2%80%94serological-reagents/LSR.md)
- **Decision Date:** May 4, 2015
- **Decision:** SESE
- **Submission Type:** Traditional
- **Regulation:** 21 CFR 866.3830
- **Device Class:** Class 2
- **Review Panel:** Microbiology
- **Attributes:** Pediatric

## Indications for Use

The EUROIMMUN Lyme ELISA (IgG/IgM) test kit is intended for the qualitative determination of IgG and/or IgM class antibodies against Borrelia burgdorferi in human serum and plasma (K⁺-EDTA, Li⁺-heparin) from symptomatic patients or people suspected of B. burgdorferi infection. It is used as an aid in the diagnosis of Lyme disease, in conjunction with other laboratory and clinical findings. All positive and borderline results should be supplemented by a second step testing method such as Western blot.

## Device Story

EUROIMMUN Lyme ELISA (IgG/IgM) is an in vitro diagnostic test kit for detecting antibodies against Borrelia burgdorferi. Input: human serum or plasma samples. Principle: Enzyme-linked immunosorbent assay (ELISA) to identify IgG/IgM class antibodies. Output: qualitative determination of antibody presence. Used in clinical laboratory settings by trained laboratory personnel. Results assist clinicians in diagnosing Lyme disease when interpreted alongside patient symptoms and other clinical findings. Positive or borderline results necessitate confirmatory testing using a second-step method like Western blot to improve diagnostic accuracy.

## Clinical Evidence

No clinical data provided in the document.

## Technological Characteristics

In vitro diagnostic ELISA test kit for qualitative antibody detection. Analyzes human serum and plasma (K2-EDTA, Li-heparin).

## 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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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

A. 510(k) Number:
K142038

B. Purpose for Submission:
To obtain a substantial equivalence determination and FDA clearance for a new device

C. Measurand:
IgG/IgM class antibodies against *Borrelia burgdorferi*

D. Type of Test:
Enzyme immunoassay

E. Applicant:
EUROIMMUN US Inc.

F. Proprietary and Established Names:
EUROIMMUN Lyme ELISA (IgG/IgM)

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 EUROIMMUN Lyme ELISA (IgG/IgM) test kit is intended for the qualitative determination of IgG and/or IgM class antibodies against *Borrelia burgdorferi* in human serum and plasma (K⁺-EDTA, Li⁺-heparin) from symptomatic patients or people suspected of *B. burgdorferi* infection. It is used as an aid in the diagnosis of Lyme disease, in conjunction with other laboratory and clinical findings. All positive and borderline results should be supplemented by a second step testing method such as Western blot.
2. Indication(s) for use: Same as Intended Use
3. Special conditions for use statement(s): N/A

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4. Special instrument requirements: N/A

I. Device Description:

The test kit contains 12 microtiter strips each with 8 break-off reagent wells coated with Borrelia burgdorferi antigens. In the first reaction step, diluted patient samples, calibrator and controls are incubated in the wells. Anti-Borrelia burgdorferi antibodies will bind to the antigens coated in the microtiter wells. The wells are washed to remove any unbound proteins and non-specific antibodies. In a second reaction step, anti-human IgG/IgM HRP enzyme conjugate (rabbit/goat) is added to each well. The enzyme conjugate will bind to any wells that have human IgG and/or IgM binding to the B. burgdorferi antigens. The wells are washed to remove any unbound HRP enzyme conjugate and 3,3,5,5 tetramethylbenzidine (TMB) enzyme substrate is added. If the HRP enzyme is present in the well (positive reaction), the HRP enzyme will react with the TMB substrate and produce a blue color. After an additional incubation time to allow the color development, a stop solution is added which turns the blue color yellow and inhibits further color development to allow for a stable spectrophotometric reading. The test strips are placed in a microplate reader and the optical density of the color is measured. The amount of antigen specific bound antibody is proportional to the color intensity.

J. Substantial Equivalence Information:

1. Predicate device name(s): Immunetics® C6 B. burgdorferi (Lyme) ELISA™
2. Predicate 510(k) number(s): K003754
3. Comparison with predicate:

Similarities

|  Item | Device | Predicate  |
| --- | --- | --- |
|  Intended Use | Detection of IgG and IgM antibodies to Borrelia burgdorferi | Same  |
|  Technology | ELISA | Same  |
|  Assay Platform | 96-well microtiter plates | Same  |
|  Conjugate | Anti-human IgG/IgM labelled with horseradish peroxidase | Same  |
|  Substrate | TMB | Same  |
|  Wash Buffer | 10x concentrate | Same  |
|  Stop Solution | 0.5 M/1 N sulphuric acid | Same  |
|  Reagent Preparation | Reagents, calibrator & controls are ready for use, except for the wash buffer. | Same  |
|  Procedure | Sample incubation with micro-well antigen coated plate, followed by a wash step, incubation with an anti-human IgG/IgM enzyme conjugate; wash step, incubation with substrate; stopping of the reaction with stop solution, photometric reading. | Same  |

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Differences

|  Item | Device | Predicate  |
| --- | --- | --- |
|  Antigen | B. burgdorferi sensu stricto B31 US strain VlsE and OspC antigens | Synthetic peptide (VlsE protein derived)  |
|  Assay Format | Qualitative | “Presumptive detection” as stated on FDA Indications for Use form  |
|  Samples Type | Serum or plasma (K+-EDTA, Li+-heparin) | Serum  |
|  Sample Dilution | 1:101 | 1:21  |
|  Calibrators and Controls | Prediluted. 1 calibrator (cut-off) 2 controls: 1 positive, 1 negative | Not prediluted. 1 calibrator (cut-off) 2 controls: 1 positive, 1 negative  |
|  Reported Results | Ratio | Index  |
|  Cut off levels | Ratio Result ≥0.8 to <1.1 borderline ≥1.1 positive | Index Result 0.91 to 1.09 equivocal ≥1.10 positive  |

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

L. Test Principle: Enzyme Immunoassay

M. Performance Characteristics (if/when applicable):

1. Analytical performance:

a. Precision/Reproducibility:

Precision: Repeatability was investigated using samples with values at different concentrations. The intra-assay repeatability is based on 20 determinations and the inter-assay repeatability is based on 40 determinations performed in 20 different runs on 10 different days (with 2 runs per day and 2 replicates per run). The results are shown below.

|  Sample No. | Mean Ratio | Within-Run |   | Within-Day |   | Between-Days |   | Total  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |  SD | %CV | SD* | %CV | SD | %CV | SD | %CV  |
|  1 | 0.1 | 0.006 | 4.1 | 0.00 | 0.0 | 0.009 | 6.6 | 0.01 | 7.8  |
|  2 | 0.2 | 0.008 | 3.9 | 0.00 | 0.0 | 0.007 | 3.5 | 0.01 | 5.2  |
|  3 | 0.5 | 0.022 | 4.2 | 0.00 | 0.0 | 0.031 | 6.0 | 0.04 | 7.3  |
|  4 | 0.8 | 0.051 | 6.4 | 0.00 | 0.0 | 0.051 | 6.4 | 0.07 | 9.1  |
|  5 | 1.1 | 0.029 | 2.5 | 0.00 | 0.0 | 0.058 | 5.2 | 0.06 | 5.8  |
|  6 | 1.9 | 0.065 | 3.5 | 0.00 | 0.0 | 0.133 | 7.2 | 0.15 | 8.0  |
|  7 | 3.0 | 0.103 | 3.4 | 0.00 | 0.0 | 0.223 | 7.4 | 0.25 | 8.2  |
|  8 | 3.7 | 0.113 | 3.0 | 0.00 | 0.0 | 0.247 | 6.6 | 0.27 | 7.3  |

*When the estimate of within-day SD is negative, it is set to 0 [CLSI EP-5: Evaluation of Precision Performance of Quantitative Measurements Methods, Repeatability Estimate.]

Reproducibility: Reproducibility was investigated using samples with values at different concentrations, which are based on 30 determinations performed with 3

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different lots with 1 run per lot and 10 replicates per run according to the package insert. Site-to-site testing was done in 48 determinations per sample performed at 3 different sites for 4 days with 2 runs per day and 2 replicates per run. The results are shown below.

|  Sample No. | Mean Ratio | Within-Run |   | Within-Day |   | Between-Days |   | Between-Sites |   | Total  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |  SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV  |
|  1 | 0.6 | 0.056 | 8.7 | 0.014 | 2.1 | 0.014 | 2.1 | 0.020 | 3.0 | 0.030 | 4.6  |
|  2 | 0.8 | 0.047 | 6.0 | 0.074 | 9.4 | 0.074 | 9.4 | 0.051 | 6.5 | 0.057 | 7.3  |
|  3 | 1.0 | 0.099 | 9.8 | 0.017 | 1.7 | 0.017 | 1.7 | 0.094 | 9.3 | 0.070 | 6.9  |
|  4 | 5.9 | 0.367 | 6.2 | 0.587 | 9.9 | 0.587 | 9.9 | 0.138 | 2.3 | 0.364 | 6.2  |
|  5 | 6.4 | 0.268 | 4.2 | 0.769 | 12.0 | 0.769 | 12.0 | 0.227 | 3.5 | 0.421 | 6.6  |
|  6 | 7.1 | 0.218 | 3.1 | 0.941 | 13.2 | 0.941 | 13.2 | 0.310 | 4.3 | 0.490 | 6.9  |
|  7 | 9.2 | 0.373 | 4.0 | 1.136 | 12.3 | 1.136 | 12.3 | 0.307 | 3.3 | 0.605 | 6.5  |
|  8 | 10.1 | 0.606 | 6.0 | 1.182 | 11.7 | 1.182 | 11.7 | 0.316 | 3.1 | 0.701 | 6.9  |

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

Analytical Specificity Study: The levels of anti-Borrelia burgdorferi antibodies from asymptomatic populations were analyzed with the EUROIMMUN Lyme ELISA (IgG/IgM) in a panel of 98 samples from an endemic region (Pennsylvania; 89 men and 9 women; age range: 18 - 56 y) and in another panel of 100 samples from a non-endemic region (Tennessee; 82 men and 18 women; age range: 2 -76 y). The results are shown below.

|  Sample Type | N | Negative | Equivocal | Positive | % Equivocal + Positive  |
| --- | --- | --- | --- | --- | --- |
|  Endemic | 98 | 95 | 0 | 3 | 3.1%  |
|  Non-endemic | 100 | 95 | 3 | 2 | 5.0%  |

Cross Reactivity: Cross reactivity was investigated using 886 serologically characterized sera positive for antibodies against different disease conditions and the results obtained are shown in the table below.

|  No. | Panel | n | Lyme ELISA (IgG/IgM)  |   |
| --- | --- | --- | --- | --- |
|   |   |   |  Negative | %  |
|  1 | Anti-Treponema pallidum | 88 | 87* | 98.9  |
|  2 | Anti-Adenovirus | 50 | 50 | 100.0  |
|  3 | Anti-Bordetella pertussis toxin | 50 | 50 | 100.0  |
|  4 | Anti-Bordetella FHA | 50 | 50 | 100.0  |
|  5 | Anti-CMV | 50 | 50 | 100.0  |
|  6 | Anti-EBV-CA | 50 | 47 | 94.0  |
|  7 | Anti-Helicobacter pylori | 50 | 50 | 100.0  |

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|  8 | Anti-HSV-1 | 50 | 50 | 100.0  |
| --- | --- | --- | --- | --- |
|  9 | Anti-Influenza virus type A | 50 | 50 | 100.0  |
|  10 | Anti-Influenza virus type B | 50 | 50 | 100.0  |
|  11 | Anti-Measles virus | 50 | 50 | 100.0  |
|  12 | Anti-Mumps virus | 50 | 50 | 100.0  |
|  13 | Anti-Mycoplasma pneumoniae | 50 | 50 | 100.0  |
|  14 | Anti-Parainfluenza virus types 1-4 | 50 | 50 | 100.0  |
|  15 | Anti-RSV | 50 | 50 | 100.0  |
|  16 | Anti-Parvovirus B19 | 35 | 34 | 97.1  |
|  17 | ANA | 14 | 14 | 100.0  |
|  18 | Rheumatoid arthritis | 17 | 16 | 94.1  |
|  19 | Fibromyalgia | 18 | 18 | 100.0  |
|  20 | Multiple sclerosis and other neurological diseases | 22 | 22 | 100.0  |

*1 Borderline sample counted as positive.

Specimens from tick-borne relapsing fever, rickettsial diseases, ehrlichiosis, babesiosis, and leptospirosis have not been tested; therefore the performance of this device is unknown with these pathologies.

**Interferences:** Hemolytic, lipemic and icteric samples showed no influence on the result up to a concentration of 1000 mg/dl for hemoglobin, 2000 mg/dl for triglycerides and 40 mg/dl for bilirubin in testing with the EUROIMMUN Lyme ELISA (IgG/IgM). Interferences with albumin, intralipids and cholesterol on the assay have not been investigated.

**f. Assay cut-off:**

**Determination of Assay Cut-off:** The assay cut-off recommendation is based on a ROC analysis from the results of 81 reference samples obtained from the Centers for Disease Control and Prevention in Atlanta, GA.

**2. Comparison studies:**

**a. Method comparison with predicate device:**

**Method Comparison Study:** A prospective study was performed with clinical samples collected from various locations in the Northeastern US. The samples were tested with the EUROIMMUN Lyme ELISA (IgG/IgM) in parallel with the predicate device. The panel consisted of 173 men, 243 women and 2 unknowns with the age range from 19 - 76 years. The table below shows the results from the prospective studies.

|  n = 418 | Predicate ELISA  |   |   |   |
| --- | --- | --- | --- | --- |
|   |   |  Positive | Equivocal | Negative  |
|  EUROIMMUN ELISA | Positive | 103 | 2 | 20  |
|   |  Borderline | 4 | 0 | 11  |
|   |  Negative | 2 | 2 | 274  |

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Positive Agreement = 96.5% (109/113) 95% C.I. 91.2 - 99.0%

Negative Agreement = 89.8% (274/305) 95% C.I. 85.9 - 93.0%

Note: Borderline/equivocal counted as positives

## b. Matrix comparison:

Serum/Plasma Comparison: The usability of plasma was investigated using sample pairs each of serum and corresponding plasma (K⁺-EDTA, Li⁺-heparin). Passing-Bablok regression was calculated for the comparison of serum to plasma. The regression equation is near the ideal correlation (intercept 0; slope 1.0) indicating equivalence of concentrations between serum and the corresponding plasma matrices. Coefficients of determination were found to be above 0.975 and % recovery compared to serum was in the range of 87 to 109 % (serum = 100 %).

|   | K⁺-EDTA Plasma | Li⁺-Heparin Plasma  |
| --- | --- | --- |
|  N | 20 | 20  |
|  Concentration Range (serum) | Ratio 0.4 – 4.3 | Ratio 0.4 – 4.3  |
|  Concentration Range (plasma) | Ratio 0.4 – 4.2 | Ratio 0.4 – 4.2  |
|  Regression Equation (y = plasma, x = serum) | y = -0.00 + 0.97 x | y = 0.04 + 0.96 x  |
|  95% C.I. of Intercept | -0.05 – 0.05 | -0.03 – 0.07  |
|  95% C.I. of Slope | 0.92 – 1.01 | 0.94 – 1.00  |
|  Coefficient of Determination R² | 0.9944 | 0.9961  |
|  Mean %Recovery | 97 % | 100 %  |
|  Range of %Recovery | 87 – 103 % | 93 – 109 %  |

## 3. Clinical studies:

### a. Clinical Sensitivity:

Sensitivity Study: A study, consisting of 100 clinically characterized Lyme disease specimens, was conducted at the manufacturer's site with the EUROIMMUN Lyme ELISA (IgG/IgM) test device in parallel with the predicate device. These specimens contain samples from early, early disseminated and late phases of the disease. The panel consisted of 36 men, 52 women and 12 unknowns. The age ranged from 16 - 80 years.

|  Disease Stage | n | EUROIMMUN Lyme ELISA (IgG/IgM) |   | Predicate ELISA  |   |
| --- | --- | --- | --- | --- | --- |
|   |   |  Positive or Borderline | Sensitivity (%) 95% C.I. | Positive or Borderline | Sensitivity (%) 95% C.I.  |
|  Acute (EM or culture positive, <3 months after onset) | 46 | 44 | 95.7 85.2 - 99.5% | 35 | 76.1 61.2 - 87.4%  |
|  Convalescent (EM or culture positive, 3-12 months after onset) | 30 | 27 | 90.0 73.5 - 97.9% | 25 | 83.3 65.3 - 94.4%  |

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|  Late
(Lyme disease with presentations other than EM, onset unknown or >1 year) | 24 | 24 | 100.0
85.8 - 100.0% | 22 | 91.7
73.0 - 99.0%  |
| --- | --- | --- | --- | --- | --- |
|  Total | 100 | 95 | 95.0
88.7 - 98.4% | 82 | 82.0
73.1 - 89.0%  |

CDC Panel Testing: Forty (40) samples of various reactivity were acquired from the Centers for Disease Control and Prevention in Atlanta, GA and evaluated internally. Of the 40 samples, 5 samples were from normal blood donors and 35 samples were from patients diagnosed with Lyme disease (clinically characterized borreliosis stratified by disease stage). All samples were tested with the EUROIMMUN Lyme ELISA (IgG/IgM) in parallel with the predicate device. Note: The results of the testing are presented here as a means of conveying further information on the performance of this assay with a characterized serum panel and does not imply an endorsement of the assay by the CDC.

Agreement to clinical diagnosis:

|  Disease Stage | n | EUROIMMUN Lyme ELISA (IgG/IgM) |   | Predicate ELISA  |   |
| --- | --- | --- | --- | --- | --- |
|   |   |  Positive or Borderline | Agreement with Clinical Diagnosis | Positive or Borderline | Agreement with Clinical Diagnosis  |
|  Normals | 5 | 0 | 100.0% | 0 | 100.0%  |
|  < 1 month | 6 | 6 | 100.0% | 6 | 100.0%  |
|  > 1 - 3 months | 11 | 10 | 90.9% | 10 | 90.9%  |
|  > 3 - 12 months | 11 | 9 | 81.8% | 9 | 81.8%  |
|  > 12 months | 7 | 7 | 100.0% | 7 | 100.0%  |
|  Total | 40 | 32 | 80.0% | 32 | 80.0%  |

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:

The range of values and positivity of different populations among different studies with the EUROIMMUN Lyme ELISA (IgG/IgM) test kit are presented below with available patient demographics.

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|  Population | n | Sex | Age Range | Ratio Results |   |   | Qualitative Results  |   |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |   |   |  Mean | Range | Std. Dev. | Positive or Borderline | %  |
|  Prospective Study | 418 | 173 men, 243 women, 2 unknown | 19-76 y; 2 unknown | 1.5 | 0.0 - 9.8 | 2.29 | 140 | 33.5%  |
|  Sensitivity Study | 100 | 36 men, 52 women, 12 unknown | 16-80 y; 12 unknown | 5.1 | 0.2 - 10.7 | 3.38 | 95 | 95.0%  |
|  CDC Panel (Lyme Disease) | 35 | unknown | unknown | 4.2 | 0.3 - 8.5 | 2.79 | 32 | 80.0%  |
|  Normal Endemic | 98 | 89 men, 9 women | 18-56 y | 0.4 | 0.1 - 5.8 | 0.85 | 3 | 3.1%  |
|  Normal Non-Endemic | 100 | 82 men, 18 women | 2-76 y | 0.3 | 0.1 - 3.8 | 0.40 | 5 | 5.0%  |

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 Part 809.10.

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/K142038](https://fda.innolitics.com/submissions/MI/subpart-d%E2%80%94serological-reagents/LSR/K142038)

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