MYCOPLASMA IGG

K033064 · Trinity Biotech USA · LON · Nov 26, 2003 · Microbiology

Device Facts

• Record ID: K033064
• Device Name: MYCOPLASMA IGG
• Applicant: Trinity Biotech USA
• Product Code: LON · Microbiology
• Decision Date: Nov 26, 2003
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.1645
• Device Class: Class 2

Intended Use

The Trinity Biotech Captia™ Mycoplasma IgG ELISA kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for semi-quantitative or qualitative determination of IgG antibodies in human serum to Mycoplasma pneumoniae for the determination of immunological experience. The Mycoplasma IgG ELISA kit may be used to evaluate paired sera for seroconversions and the presence of a significant increase in specific IgG as and aid in the diagnosis of Mycoplasma pneumoniae infection in the adult population.

Device Story

The Captia Mycoplasma IgG ELISA is an in vitro diagnostic test for human serum. It utilizes purified Mycoplasma pneumoniae antigen immobilized on microtiter wells. Patient serum is added; if specific IgG antibodies are present, they bind to the antigen. After washing, enzyme-labeled anti-human IgG is added, followed by a substrate solution. The resulting color change is measured photometrically, providing an indirect measurement of antibody concentration. The test is performed in clinical laboratories by trained technicians. Results are used by physicians to aid in the diagnosis of Mycoplasma pneumoniae infection by identifying seroconversion or significant antibody increases in paired acute and convalescent serum samples. The device provides semi-quantitative or qualitative results to assess patient immunological experience.

Clinical Evidence

No clinical data provided. Device is a distributor name change for a previously cleared predicate (K971393); performance characteristics were established in the predicate submission.

Technological Characteristics

Enzyme-Linked Immunosorbent Assay (ELISA) using purified Mycoplasma pneumoniae antigen coated on solid-phase microtiter wells. Employs enzyme-labeled anti-human IgG conjugate and substrate for photometric detection. Designed for semi-quantitative or qualitative analysis. Standard laboratory equipment required for photometric measurement.

Indications for Use

Indicated for adult population to detect IgG antibodies to Mycoplasma pneumoniae in human serum to determine immunological experience and aid in diagnosis of infection via evaluation of paired sera for seroconversion or significant IgG increase.

Regulatory Classification

Identification

A fully automated short-term incubation cycle antimicrobial susceptibility system is a device that incorporates concentrations of antimicrobial agents into a system for the purpose of determining in vitro susceptibility of bacterial pathogens isolated from clinical specimens. Test results obtained from short-term (less than 16 hours) incubation are used to determine the antimicrobial agent of choice to treat bacterial diseases.

Special Controls

*Classification.* Class II (special controls). The special control for this device is FDA's guidance document entitled “Class II Special Controls Guidance Document: Antimicrobial Susceptibility Test (AST) Systems; Guidance for Industry and FDA.”

Predicate Devices

• Mycoplasma IgG ELISA Test Kit (K971393)

Submission Summary (Full Text)

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K033064

Summary of Safety and Effectiveness Information Mycoplasma IgG ELISA Test Kit

- I. Trinity Biotech 2823 Girts Road Jamestown, NY 14701 Contact person: Bonnie B. DeJoy Telephone: [phone] Date of preparation: Nov. 20, 2003
- II. Description of Device

The Mycoplasma IgG ELISA kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for semi-quantitative or qualitative determination of IgG antibodies in human serum to Mycoplasma pneumoniae for the determination of immunological experience. The Mycoplasma IgG ELISA kit may be used to evaluate paired sera for the presence of seroconversions and a significant increase in specific IgG as an aid in the diagnosis of Mycoplasma pneumoniae infection in the adult population. For In Vitro Diagnostic Use Only.

The Mycoplasma IgG ELISA test is an enzyme linked immunosorbent assay to detect IgG antibodies to Mycoplasma pneumoniae. Purified Mycoplasma pneumoniae antigen is attached to a solid phase microtiter well. Diluted test sera are added to each well. If the antibodies are present that recognize the antigen, they will bind to the antigen in the well. After incubation, the wells are washed to remove unbound antibody. An enzyme labeled anti-human IgG is added to each well. After incubation, the wells are washed to remove unbound conjugate. A substrate solution is added to each well. If enzyme is present, the substrate will undergo a color change. After an incubation period the reaction is stopped and the color intensity is measured photometrically, producing and indirect measurement of specific antibody in the patient specimen.

- III. Predicate Device
The Mycoplasma IgG ELISA test is substantially equivalent to the IFA test. Equivalence is demonstrated by the following comparative results:

# Performance Characteristics

### % Agreement Positive and % Agreement Negative

Two different sites compared the Trinity Biotech Mycoplasma IgG ELISA test relative to a commercial IFA kit. The two sites were R&D laboratories at commercial companies located in Maryland and New York and affiliated with the manufacturer of the kit. The 187 frozen retrospective sera from the first study were from normal individuals of various ages, gender, from Lyme disease endemic and non-endemic areas. The results of the first study are summarized in Table 2. The 176 frozen retrospective sera from the second study were randomly selected sera from normal inividuals of various ages, gender, and geographical location. The results of the second study are summarized in Table 3. None of the performance characteristics were established with specimens from patients having documented mycoplasma infections.

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### Table 2 Comparison of Trinity Biotech Mycoplasma IgG ELISA and IFA Study 1

### Trinity Biotech Mycoplasma IgG ELISA

| | + | eq | - | Total |
|--------------------------|-----|----|----|-------|
| + | 117 | 13 | 6 | 136 |
| Mycoplasma<br>IFA (1:32) | 20* | 6 | 25 | 51 |
| Total | 137 | 19 | 31 | 187 |

| % Agreement positive = 117/123 = 95.1% | 95% Confidence interval = 91.2% - 99.0% |
|----------------------------------------|-----------------------------------------|
| % Agreement negative = 25/45 = 55.6% | 95% Confidence interval = 40.7% - 70.4% |
| % Agreement = 142/168 = 84.5% | 95% Confidence interval = 78.19 - 90.1% |

* All 20 sera were found to be positive by an alternate ELISA. Equivocals were not included in the above calculations. The 95% Confidence Intervals were calculated using the normal method.

### Table 3 Comparison of Trinity Biotech Mycoplasma IgG ELISA and IFA Study 2

### Trinity Biotech Mvcoplasma IgG ELISA

| | + | eq | - | Total | |
|--------------------------|-------|-----|---|-------|-----|
| + | 132 | 5 | 2 | 139 | |
| Mycoplasma<br>IFA (1:32) | - | 20* | 0 | 17 | 37 |
| | Total | 152 | 5 | 19 | 176 |

| % Agreement positive = 132/134 = 98.5% | 95% Confidence interval = 96.4% - 100.0% |
|----------------------------------------|------------------------------------------|
| % Agreement negative = 17/37 = 45.9% | 95% Confidence interval = 29.6% - 62.3% |
| % Agreement = 149/171 = 87.1% | 95% Confidence interval = 82.0% - 92.3% |

* All 20 sera were found to be positive by an alternate ELISA.

Equivocals were not included in the above calculations.

The 95% Confidence Intervals were calculated using the normal method.

Please be advised that "% agreement positive" and "% agreement negative" refer to the comparison of this assay's results to that of a similar assay. There was not an attempt to correlate the assay's results with disease presence or absence. No judgment can be made on the comparison assay's accuracy to predict disease.

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### Precision

Seven sera were assayed ten times each on three different assays at two different sites. Both sites were affiliated with the manufacturer of the kit. The intra- and inter-assay precision at each site is shown in Tables 4 and 5. The inter-site precision is shown in Table 6. With appropriate technique the user should obtain precision of < 15% CV.

| Mycoplasma IgG ELISA Intra- and Inter-Assay Precision Study 1 | | | | |
|---------------------------------------------------------------|---------------------------|---------------------------|---------------------------|------------------------------|
| Sera# | Assay 1 (n=10)<br>X SD CV | Assay 2 (n=10)<br>X SD CV | Assay 3 (n=10)<br>X SD CV | Inter-Assay(n=30)<br>X SD CV |
| 1 | 0.42 0.054 12.86% | 0.36 0.033 9.18% | 0.47 0.025 5.48% | 0.42 0.054 13.02% |
| 2 | 0.29 0.043 14.58% | 0.23 0.026 11.38% | 0.29 0.034 11.91% | 0.27 0.043 15.93% |
| 3 | 3.54 0.274 7.73% | 3.24 0.244 7.53% | 3.50 0.273 7.78% | 3.43 0.274 7.98% |
| 4 | 1.89 0.133 7.05% | 1.76 0.142 8.09% | 1.90 0.103 5.42% | 1.85 0.133 7.21% |
| 5 | 0.42 0.059 13.93% | 0.33 0.051 15.21% | 0.42 0.044 10.45% | 0.39 0.059 15.02% |
| 6 | 1.09 0.103 9.49% | 1.03 0.088 8.56% | 1.16 0.096 8.28% | 1.09 0.103 9.45% |
| 7 | 2.31 0.218 9.44% | 2.21 0.286 12.98% | 2.41 0.160 6.66% | 2.31 0.218 9.46% |
| HPC | | | | 4.51 0.078 1.72%* |
| Cal | | | | 2.50 0.072 2.89%** |
| LPC | | | | 1.31 0.135 10.33% |
| NC | | | | 0.49 0.049 10.14% |
| * n = 3 | | | | |
| ** n = 9 | | | | |

| Table 4 | | | | | |
|---------------------------------------------------------------|--|--|--|--|--|
| Mycoplasma IgG ELISA Intra- and Inter-Assay Precision Study 1 | | | | | |

| Table 5 | | | | | |
|---------------------------------------------------------------|--|--|--|--|--|
| Mycoplasma IgG ELISA Intra- and Inter-Assay Precision Study 2 | | | | | |

| | | Assay 1 (n=10) | | | Assay 2 (n=10) | | | Assay 3 (n=10) | | | Inter-Assay(n=30) | | |
|---------|--|----------------|-------|--------|----------------|-------|--------|----------------|-------|--------|-------------------|-------|--------|
| Sera# | | X | SD | CV | X | SD | CV | X | SD | CV | X | SD | CV |
| 1 | | 0.23 | 0.013 | 5.74% | 0.22 | 0.024 | 10.76% | 0.25 | 0.028 | 11.04% | 0.24 | 0.025 | 10.75% |
| 2 | | 0.17 | 0.022 | 12.71% | 0.17 | 0.023 | 13.58% | 0.19 | 0.018 | 9.45% | 0.18 | 0.023 | 12.90% |
| 3 | | 3.58 | 0.199 | 5.56% | 3.58 | 0.161 | 4.51% | 3.70 | 0.154 | 4.17% | 3.62 | 0.176 | 4.87% |
| 4 | | 1.84 | 0.102 | 5.57% | 1.84 | 0.173 | 9.41% | 2.07 | 0.135 | 6.55% | 1.91 | 0.174 | 9.07% |
| 5 | | 0.35 | 0.016 | 4.54% | 0.33 | 0.028 | 8.37% | 0.37 | 0.029 | 7.90% | 0.35 | 0.029 | 8.29% |
| 6 | | 1.15 | 0.070 | 6.09% | 1.14 | 0.074 | 6.47% | 1.27 | 0.078 | 6.15% | 1.19 | 0.092 | 7.78% |
| 7 | | 2.22 | 0.147 | 6.64% | 2.16 | 0.154 | 7.13% | 2.31 | 0.105 | 4.56% | 2.23 | 0.148 | 6.63% |
| HPC | | | | | | | | | | | 3.48 | 0.191 | 5.48%* |
| Cal | | | | | | | | | | | 2.51 | 0.056 | 2.21%* |
| LPC | | | | | | | | | | | 1.43 | 0.140 | 9.78% |
| NC | | | | | | | | | | | 0.13 | 0.03 | 23.08% |
| * n = 3 | | | | | | | | | | | | | |

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# Table 6 Trinity Biotech Mycoplasma IgG ELISA Inter Site Precision Study

| Inter-Assay | | | | |
|-------------|------|-------|--------|----|
| Sera# | X | SD | CV | n |
| 1 | 0.33 | 0.101 | 30.89% | 60 |
| 2 | 0.22 | 0.057 | 25.73% | 60 |
| 3 | 3.52 | 0.247 | 7.01% | 60 |
| 4 | 1.88 | 0.157 | 8.33% | 60 |
| 5 | 0.37 | 0.050 | 13.43% | 60 |
| 6 | 1.14 | 0.108 | 9.50% | 60 |
| 7 | 2.27 | 0.189 | 8.34% | 60 |
| HPC | 4.00 | 0.581 | 14.53% | 6 |
| CAL | 2.51 | 0.063 | 2.50% | 18 |
| LPC | 1.37 | 0.140 | 10.24% | 6 |
| NC | 0.31 | 0.199 | 64.46% | 6 |

A total of 456 determinations were made at the two sites. The only specimen to change status was # 6 which was positive 38 times and equivocal 22 times.

X = Mean SD = Standard Deviation CV = Coefficient of Variation = SD/X x 100

The methods in NCCLS EP5 were utilized for precision parameters.

# Linearity

### Simulated Paired Sera Evaluation

To evaluate the linearity of the assay 20 positive sera were serially two-fold diluted and run on the assay. The ISR values were compared to log2 of dilution by standard linear regression. The r values were all ≥0.974. The data indicate that the antibody can be semi-quantitated by using a single serum dilution. The detection of a significant antibody increase may be made only by an evaluation of paired specimens, acute and convalescent. To validate the % agreement positive of the paired sera procedure, the percent rise in ISR value was calculated for 56 pairs that had a four-fold dilution where the acute sera had a value of less than 2.18. All 56 pairs demonstrated a >46% rise in ISR value, showing a significant rise in antibody. Therefore, the paired sera procedure demonstrated 100% agreement positive in being able to detect a four-fold increase in antibody level when the acute sera has a value of <2.18.

Figure 1 illustrates the linearity of a representative serum. The ISR values were compared to log> of dilution by standard linear regression

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Image /page/4/Figure/0 description: The image is a graph titled "Linearity of Mycoplasma ELISA". The graph shows the ISR value on the y-axis and the dilution factor on the x-axis. The dilution factors are Neat, 1:2, 1:4, 1:8, and 1:16. The graph also shows that r = 0.989, slope = 0.66, and y inter = -0.24.

### Complement Fixation Paired Serum Study

Eleven serum pairs tested by CF from patients suspected of having acute Mycoplasma pneumoniae infection were assayed on the Trinity Biotech Mycoplasma IgG ELISA assay. Each serum pair was evaluated to determine a significant rise in antibody. Four serum pairs could not be used due to the acute serum being too high. The remaining seven pairs all demonstrated a >46% rise in ISR values thus giving a 100% agreement positive versus CF for showing a significant rise in antibody for serum meeting the paired sera criteria.

### Reproducibility Study

Fifty different sera with various levels of activity were assayed at three different sites. Two sites were R&D laboratories at commercial companies located in Maryland and New York. The third site was a large clinical laboratory located in Pennsylvania. The data from the three sites show good correlation with Pearson product moment correlation coefficients of >0.987 between the sites. Excluding equivocals (n = 4) one determination varied from its expected result (negative result for a positive specimen) giving a percent agreement of expected results between the three sites of 99.3% (145/146). The expected results were derived from previous Trinity Biotech ELISA testing of the samples. Three sera changed status in this study: one serum was equivocal at one site and negative at the other two sites; the second serum was equivocal at two sites and positive at the third; and the third serum was positive at one site, equivocal at second site, and negative at the third site.

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Image /page/5/Picture/1 description: The image is a black and white seal for the Department of Health & Human Services - USA. The seal is circular with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the perimeter. Inside the circle is a stylized image of an eagle with three lines representing its wings and a wavy line representing its tail.

NOV 26 2003

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Ms. Bonnie B. DeJoy Director, Quality Systems Trinity Biotech USA P.O. Box 1059 Jamestown, NY 14702-1059

Re: k033064

> Trade/Device Name: Captia Mycoplasma IgG ELISA Regulation Number: 21 CFR 866.3375 Regulation Name: Mycoplasma spp. serological reagents Regulatory Class: Class II Product Code: LON Dated: May 14, 2003 Received: May 15, 2003

Dear Ms. DeJoy:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration. Iisting 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 (PMA), it may be subject to such additional controls. Existing major regulations affecting your device can be found in Title 21, Code of Federal Regulations (CFR), Parts 800 to 895. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).

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Page 2 -

This letter will allow you to begin marketing your device as described in your Section 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 information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers, International and Consumer Assistance at its toll-free number [phone] or [phone] or at its Internet address http://www.fda.gov/cdrh/dsma/dsmamain.html.

Sincerely yours,

Steven Sutman

Steven I. Gutman, M.D., M.B.A. Director Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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510(k) Number: K033064

Device Name: Trinity Biotech Captia™ Mycoplasma IgG ELISA

Indications For Use: The Trinity Biotech Captia™ Mycoplasma IgG ELISA kit is an Enzyme-Linked Immunosorbent Assay (ELISA) for semi-quantitative or qualitative determination of IgG antibodies in human serum to Mycoplasma pneumoniae for the determination of immunological experience. The Mycoplasma IgG ELISA kit may be used to evaluate paired sera for seroconversions and the presence of a significant increase in specific IgG as and aid in the diagnosis of Mycoplasma pneumoniae infection in the adult population.

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Concurrence of CDRH, Office of Device Evaluation (ODE)

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510(k) K033064