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 Description

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.

AI/ML Overview

Here's an analysis of the provided text, focusing on the acceptance criteria and the study proving the device meets them:

1. Table of Acceptance Criteria and Reported Device Performance

Performance Characteristic	Acceptance Criteria (Implicit)	Reported Device Performance
Agreement with Predicate Device (IFA)
% Agreement Positive (Study 1)	High positive agreement (e.g., >90%)	95.1% (95% CI: 91.2% - 99.0%)
% Agreement Negative (Study 1)	Sufficient negative agreement (no explicit target, but acceptable for predicate comparison)	55.6% (95% CI: 40.7% - 70.4%)
% Overall Agreement (Study 1)	High overall agreement (e.g., >80%)	84.5% (95% CI: 78.1% - 90.1%)
% Agreement Positive (Study 2)	High positive agreement (e.g., >90%)	98.5% (95% CI: 96.4% - 100.0%)
% Agreement Negative (Study 2)	Sufficient negative agreement (no explicit target, but acceptable for predicate comparison)	45.9% (95% CI: 29.6% - 62.3%)
% Overall Agreement (Study 2)	High overall agreement (e.g., >80%)	87.1% (95% CI: 82.0% - 92.3%)
Precision (CV)	< 15% CV (guidance for user)
Intra-Assay Precision (Study 1)	< 15% CV (observed)	Ranged from 5.48% to 14.58% (Individual assay/sera)
Inter-Assay Precision (Study 1)	< 15% CV (observed)	Ranged from 7.21% to 15.93% (Individual sera)
Intra-Assay Precision (Study 2)	< 15% CV (observed)	Ranged from 4.17% to 13.58% (Individual assay/sera)
Inter-Assay Precision (Study 2)	< 15% CV (observed)	Ranged from 4.87% to 12.90% (Individual sera)
Inter-Site Precision (Table 6 Samples 3-7, HPC, CAL, LPC)	< 15% CV (observed)	Ranged from 2.50% to 14.53%
Inter-Site Precision (Table 6 Samples 1, 2, NC)	(No specific explicit criterion, but observed values for some samples are higher than 15% CV)	30.89% (Sera 1), 25.73% (Sera 2), 64.46% (NC)
Linearity (r value)	≥ 0.974 (for log2 dilution vs ISR)	All ≥ 0.974
Paired Sera Evaluation (% rise in ISR value)	> 46% rise in ISR value when acute sera < 2.18, leading to 100% agreement positive.	100% agreement positive (56/56 pairs showed > 46% rise)
CF Paired Serum Study (% rise in ISR value)	> 46% rise in ISR value for serum meeting paired sera criteria, leading to 100% agreement positive.	100% agreement positive (7/7 pairs showed > 46% rise)
Reproducibility (Pearson correlation coefficient)	> 0.987 (between sites)	> 0.987
Reproducibility (% agreement of expected results)	High agreement (e.g., >95%)	99.3% (145/146)

Note on Acceptance Criteria: The document primarily presents performance characteristics and compares them to a predicate device or implied standards (e.g., "With appropriate technique the user should obtain precision of < 15% CV"). Explicit, pre-defined numerical acceptance criteria are not always stated, but the reported values demonstrate the device's performance in relation to the predicate or common laboratory standards. For precision, the text itself suggests "< 15% CV" as an expected outcome with appropriate technique.

2. Sample Sizes Used for the Test Set and Data Provenance

Agreement with Predicate Device (IFA):
- Study 1: 187 frozen retrospective sera.
  - Provenance: Normal individuals of various ages, gender, from Lyme disease endemic and non-endemic areas. (Country of origin not specified, but commercial companies were located in Maryland and New York, affiliated with the manufacturer).
- Study 2: 176 frozen retrospective sera.
  - Provenance: Randomly selected sera from normal individuals of various ages, gender, and geographical location. (Country of origin not specified, but commercial companies were located in Maryland and New York, affiliated with the manufacturer).
Precision:
- 7 sera, each assayed 10 times on 3 different assays at 2 different sites. This means 7 * 10 * 3 * 2 = 420 individual tests for sera, plus controls.
- Provenance: Not explicitly stated for the sera samples themselves, but the testing was done at R&D laboratories at commercial companies affiliated with the manufacturer (Maryland and New York).
Linearity (Simulated Paired Sera):
- 20 positive sera, serially two-fold diluted. The number of dilutions is not specified, but at least 4-5 dilutions per serum would be typical (Neat, 1:2, 1:4, 1:8, 1:16 as shown in the graph).
- 56 paired sera (for % agreement positive detection of four-fold increase).
- Provenance: Not specified for individual sera or paired sera.
Complement Fixation Paired Serum Study:
- 11 serum pairs, with 7 ultimately used for analysis.
- Provenance: From patients suspected of having acute Mycoplasma pneumoniae infection, tested by CF. (Country of origin not specified).
Reproducibility Study:
- 50 different sera.
- Provenance: Assayed at three different sites: two R&D labs at commercial companies (Maryland and New York) and one large clinical laboratory (Pennsylvania).

3. Number of Experts Used to Establish Ground Truth for the Test Set and Qualifications

No "experts" per se were used to establish ground truth in the traditional sense for the primary agreement studies.
The ground truth for the agreement studies (Studies 1 and 2) was the Mycoplasma IFA (1:32) kit, which is the predicate device. This is a comparative study against an existing, legally marketed device, not a comparison to a clinical diagnosis established by experts.
For the Complement Fixation Paired Serum Study, the ground truth for "significant rise in antibody" was established by the CF method.
The "expected results" for the Reproducibility Study were derived from "previous Trinity Biotech ELISA testing of the samples."

4. Adjudication Method for the Test Set

For the primary agreement studies (Studies 1 and 2), the comparison was directly between the new Trinity Biotech Mycoplasma IgG ELISA and the commercial IFA kit. There was no mention of an adjudication process between different readers or methods beyond the direct comparison.
In cases where the Trinity Biotech ELISA and the IFA kit disagreed, all 20 sera in Study 1 and 20 sera in Study 2 that were positive by IFA but negative by the Trinity Biotech ELISA were further tested by "an alternate ELISA" to confirm their positive status. This serves as a form of secondary evaluation or tie-breaking for discordant results.
For the reproducibility study, "expected results were derived from previous Trinity Biotech ELISA testing." This suggests a pre-defined reference, not an adjudication of new readings.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done, and Effect Size

No, a Multi-Reader Multi-Case (MRMC) comparative effectiveness study was not done.
This device is an in vitro diagnostic (ELISA test kit), not an imaging or interpretation device that would typically involve multiple human readers interpreting results with and without AI assistance. The performance studies focus on the analytical and clinical agreement of the assay itself with a predicate device and its internal consistency (precision, linearity, reproducibility).

6. If a Standalone (Algorithm Only Without Human-in-the-Loop Performance) Was Done

Yes, the studies presented are all standalone performance evaluations of the Mycoplasma IgG ELISA kit.
The ELISA kit is a laboratory test where the "algorithm" is the biochemical reaction and photometric measurement, yielding a quantitative or semi-quantitative result. There is no human-in-the-loop variable being tested in terms of interpreting algorithmic output. The performance metrics (agreement, precision, linearity, reproducibility) represent the intrinsic performance of the device on its own.

7. The Type of Ground Truth Used

Primary Ground Truth for Agreement Studies: The predicate device (commercial IFA kit) was used as the comparator for determining "% Agreement Positive" and "% Agreement Negative." This is a comparator or reference method ground truth.
Secondary Ground Truth for Discordant Results: An "alternate ELISA" was used to confirm positive status for samples where the predicate IFA was positive but the study device was negative.
Paired Sera Studies: The "significant rise in antibody" was determined by either the calculated % rise in ISR value within the Trinity Biotech ELISA or by the Complement Fixation (CF) method as a reference.
Reproducibility Study: "Expected results" were derived from "previous Trinity Biotech ELISA testing".

8. The Sample Size for the Training Set

No explicit training set is mentioned in the description.
ELISA kits, particularly for a 510(k) submission, are typically validated through analytical and clinical performance studies, not by training a machine learning algorithm. The "development" or "optimization" would involve chemical and biological engineering, not data training in the AI sense.
The provided studies evaluate the final product's performance, not the iterative development process that might involve a 'training set' for an AI model.

9. How the Ground Truth for the Training Set Was Established

As no explicit training set for an AI/algorithm was described, the concept of establishing ground truth for a training set does not apply directly to this device's validation as presented in the summary.
The 'ground truth' pertinent to the evaluation was established by either a predicate device (IFA), an alternate ELISA, or a CF method as described in section 7.

Summary

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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: 716-483-3851 Date of preparation: Nov. 20, 2003
II. Description of Device

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
MycoplasmaIFA (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
MycoplasmaIFA (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)X SD CV	Assay 2 (n=10)X SD CV	Assay 3 (n=10)X SD CV	Inter-Assay(n=30)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 (301) 594-3084. 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 (800) 638-2041 or (301) 443-6597 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.

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

Prescription Use (Per 21 XFR 801.109)
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Over-The-Counter Use (Optional Format 1-2-96)
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Division Sign-OffOffice of In Vitro Diagnostic Device Evaluation and Safety

510(k) K033064

Regulation Number and Section

§ 866.1645 Fully automated short-term incubation cycle antimicrobial susceptibility system.

(a)
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.(b)
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.”