The RAMP Troponin I Assay is a quantitative immunochromatographic test indicated for use as an in vitro diagnostic product used with the RAMP Clinical Reader to measure cardiac troponin I levels in EDTA whole blood. Measurement of cardiac troponin I aids in the rapid diagnosis of acute myocardial infarction (AMI). The RAMP Troponin I Assay is intended to be used only to prioritize patient management for those suspected of AMI.

Device Description

The RAMP Troponin I Assay is a quantitative immunochromatographic test for the rnic Tric Troponia T-Thous) Do A whole blood. Diluted EDTA whole blood is added to the sample well of the Test Cartridge which houses the immunochromatographic test strip our pro real blood cells are retained in the sample pad, and the separated plasma migrates along the strip. Fluorescent-dyed latex particles coated with anti-Tril antibodies hightion and in the sample. As the sample migrates along the strip, Tnl bound particles are immobilized at the detection zone, and additional particles are immobilized at the internal control zone. The RAMP Reader then measures the amount of fluorescence emitted by the complexes bound at the detection zone and at the internal control zone. Using a ratio between the two fluorescence values, a quantitative reading is calculated.

AI/ML Overview

This document describes the RAMP Troponin I Assay, a quantitative immunochromatographic test for measuring cardiac troponin I levels in EDTA whole blood, intended to aid in the rapid diagnosis of acute myocardial infarction (AMI).

1. A table of acceptance criteria and the reported device performance

The document does not explicitly state pre-defined acceptance criteria for performance metrics such as precision, linearity, or analytical sensitivity. Instead, it provides performance characteristics and then concludes that the device is "substantially equivalent" to predicate devices. For a regulatory submission like a 510(k), substantial equivalence implies that the device performs as well as or better than existing legally marketed devices.

However, we can infer some criteria and list the reported performance:

Performance Characteristic	Reported Device Performance (RAMP Troponin I Assay)
Precision
Intra-assay CV	8.7% and 8.3% (at 1.05 and 5.01 ng/mL Tnl)
Total precision CV	10.0% and 8.3% (at 1.05 and 5.01 ng/mL Tnl)
Low end precision (CV ≤ 10%)	0.21 ng/mL Tnl (within run), 0.15 ng/mL Tnl (total), 0.10 ng/mL Tnl (whole blood)
Low end precision (CV ≤ 20%)	0.14 ng/mL Tnl (within run)
Linearity (R value)	0.997
Slope	1.019
Offset	0.279
Percent Recovery	95 to 115% (average 105%)
Hook Effect	No high dose hook effect up to 500 ng/mL Tnl
Lower Limit of Detection (LLD)	0.03 ng/mL Tnl
Upper Reportable Limit	> 32 ng/mL Tnl
Cross-reactivity	No significant cross-reactivity with Skeletal Troponin I, Cardiac Troponin T, Cardiac Troponin C. Minimal with HAMA, HAGA, HARA, RhF.
Interference	No evidence of cross-reactivity or interference with hemoglobin, triglyceride, bilirubin, cholesterol, heparin at very high physiological concentrations.
Method Comparison (vs. Dimension) - Combined Pop. R value	0.988

2. Sample size used for the test set and the data provenance (e.g. country of origin of the data, retrospective or prospective)

Precision Study Test Set:
- Sample Size: 184 subjects (55 normal individuals, 129 patients suspected of AMI).
- Provenance: Samples were selected from those obtained during the Method Comparison Study. The document does not explicitly state the country of origin but implies a clinical setting via "individual hospital criteria." It indicates the samples were retrospective for the precision study, as they were "stored refrigerated for up to one day between analyses" and subsequently analyzed in duplicates "over 10 days."
Method Comparison Study Test Set:
- Sample Size: 365 subjects (180 normal individuals, 185 patients suspected of AMI).
- Provenance: The samples were obtained from subjects where "EDTA and heparin whole blood samples were obtained for each of these subjects." "All normal subjects were consented. Waste samples were used for the subjects suspected of AMI." This suggests a prospective collection for this specific study for the normal subjects, and potentially retrospective or waste samples for the AMI subjects. Country of origin is not specified but the submission is to the FDA, implying US or internationally recognized standards.
Expected Values (Normal Range) Study Test Set:
- Sample Size: 180 healthy individuals.
- Provenance: "Whole blood samples from 180 healthy individuals... were assayed". This implies a prospective collection from a healthy population.

3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts (e.g. radiologist with 10 years of experience)

The studies described are for an in-vitro diagnostic device measuring a biomarker (Troponin I). The "ground truth" here is the actual concentration of Troponin I in the samples. This is typically established by reference methods or comparison to legally marketed predicate devices, not by human expert consensus or radiologists.

For the Method Comparison Study, the Dimension® RxL Cardiac Troponin-I Flex® assay (a predicate device, K973650) served as a reference for comparison, implying its results were considered a form of "ground truth" or a highly reliable comparator. The data was also compared to the Triage Cardiac Panel®, another predicate device.
For analytical performance (precision, linearity, LLD), the ground truth is established by preparing known concentrations of the analyte (Tnl antigen) in a controlled laboratory setting.

Therefore, the concept of "experts" establishing ground truth in the way described (e.g., radiologists) is not applicable to this type of device and study.

4. Adjudication method (e.g. 2+1, 3+1, none) for the test set

Adjudication methods like 2+1 or 3+1 are typically used in studies involving human interpretation (e.g., radiology reads) where discrepancies between readers need to be resolved. This document describes studies for an automated in-vitro diagnostic device measuring a biomarker. Therefore, no adjudication method of this type was used or is relevant. The device output is a quantitative measurement.

5. If a multi reader multi case (MRMC) comparative effectiveness study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

No, a multi-reader multi-case (MRMC) comparative effectiveness study was not done. This type of study focuses on the impact of a device (often AI-based) on human reader performance, which is not applicable to an automated quantitative immunoassay like the RAMP Troponin I Assay. The device provides a direct numerical measurement of a biomarker, not an interpretation that human readers would "improve with."

6. If a standalone (i.e. algorithm only without human-in-the loop performance) was done

Yes, the studies described are inherently standalone algorithm performance studies. The RAMP Troponin I Assay, in conjunction with the RAMP Reader, is an automated system that provides quantitative measurements. The performance characteristics (precision, linearity, analytical sensitivity, hook effect, cross-reactivity, interference, and method comparison) are all evaluating the device's ability to accurately measure Troponin I concentrations directly, without human interpretation of the assay's output influencing the direct measurement itself. The "human-in-the-loop" aspect for this device would be the clinician's interpretation of the numerical result in the context of patient symptoms, not the interpretation of the assay output itself.

7. The type of ground truth used (expert consensus, pathology, outcomes data, etc)

The primary "ground truth" used for this quantitative assay comprises:

Known concentrations of analyte: For analytical sensitivity, linearity, and recovery studies, known concentrations of Tnl antigen were prepared and used as the reference.
Results from legally marketed predicate devices: For the method comparison study, the Dimension® RxL Cardiac Troponin-I Flex® Assay and Triage Cardiac Panel® Troponin I Assay were used as comparators, providing a "ground truth" for clinical sample concentrations.

This is distinct from ground truth based on expert consensus, pathology, or outcomes data, which are more common for diagnostic imaging or clinical prediction models.

8. The sample size for the training set

The document does not explicitly describe a "training set" in the context of machine learning or AI models, as this is a chemical immunoassay, not a software algorithm that learns from data.

However, if we consider "training" in a broader sense related to assay development and optimization (e.g., determining assay parameters, reagent concentrations), that information is not provided as part of this 510(k) summary. The summary focuses on the final performance validation of the developed assay.

9. How the ground truth for the training set was established

As there is no "training set" described in the machine learning sense, this question is not fully applicable. For the development and establishment of the assay itself (analogous to "training" certain parameters), the ground truth would have been established through controlled laboratory experiments using purified Troponin I antigens at known concentrations and characterization against reference materials. However, details of this developmental phase are not included in the 510(k) "Summary of Studies," which focuses on performance verification.

Summary

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510(K) SUMMARY OF SAFETY AND EFFECTIVENESS 4.0

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92.

The assigned 510(k) number is: K033745

1. Establishment

Response Biomedical Corp. 8081 Lougheed Highway Burnaby, British Columbia Canada, V5A 1W9

Tel: (604) 681-4101 Fax: (604) 412-9830

William J. Radvak Contact: President and CEO

November 27, 2003 Prepared:

2. Regulatory Information

Trade Name:	Response Biomedical Corp. RAMP® Troponin I Assay
Common Name:	Troponin I immunological test system
Classification Name:	Troponin I immunological test system
Regulation Number:	862.1215
Product Code:	MMI
Panel:	Clinical Chemistry

3. Predicate Device

Immunoassay:	Triage Cardiac Panel®; Troponin I Assay (K973126) which iscurrently being marketed by Biosite Diagnostics, Inc.
Immunoassay:	Dimension® RxL Cardiac Troponin-I Flex®, (K973650) whichis currently being marketed by Dade Behring Inc.

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4. Description of the Device

The RAMP Reader then measures the amount of fluorescence emitted by the complexes bound at the detection zone and at the internal control zone. Using a ratio between the two fluorescence values, a quantitative reading is calculated.

5. Indication for Use

6. Comparison of Technological Characteristics

The RAMP Troponin I Assay, Triage Cardiac Panel (Triage) Troponin I, and Dade Dimension RxL (Dimension) Cardiac Troponin-I (Troponin I) Flex Assays are for the quantitative measurement of Tnl in human whole blood (RAMP and Triage) or plasma (Triage and Dimension). All three immunoassays utilize the binding of Tnl to specific antibodies and utilize light in their respective detection systems. Both the RAMP and Triage assays measure light production from a fluorescence reaction using a fluorometer while the Dimension measures the amount of colored product produced which is directly proportional to the concentration of Tnl present in the patient sample. Both the RAMP Troponin I and the Triage Troponin I assays are quantitative immunochromatographic tests, whereas the Dimension Troponin I test is a sandwich enzyme immunoassay.

7. Summary of Studies

PERFORMANCE CHARACTERISTICS

PRECISION: The intra-assay and the inter-assay precision of the RAMP Troponin I Assay were determined by one operator assaying duplicates of two standards (1.05 and 5.01 ng/mL Tnl) twice each day over 10 days. The mean, standard deviation and %CV were calculated for the predicted Tnl at each concentration. Within run precision was 8.7% and 8.3% respectively. Total precision was 10.0% and 8.3% respectively. Low end precision was also determined by the same method using Plasma Standards of 0.09, 0.15, 0.22, 0.29, 0.40 and 0.70 ng/mL Tnl. The assay concentration in standards where 10.0% or less CV is achieved was 0.21 ng/mL Tnl and where 20.0% or less CV is achieved was 0.14

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ng/mL Tnl for within run precision. The assay concentration in standards where 10.0% or ngmic Yil for within full problem. The as a viere 20.0% or less CV is achieved was 0.15 ng/mL Tnl for total precision. Low end precision of the RAMP Troponin I Assay was 0.15 Ilgmir Thi for total prodision: Low one proclaired assaying 10 replicates of each of also determined in ED World Color assay concentration in blood where 10.0% or less CV is 5 spired ini concentrations. The assure of is achieved was 0.10 ng/mL Tnl.

LINEARITY and PERCENT RECOVERY: Tnl antigen concentrations of 0.86, 1.72, 3.44, 6.88, 13.75, and 27.5 ng/mL were prepared in normal donor EDTA blood. The linearity 0.00, 15.15, and 27.5 hgmd. Were propan assaying five replicates of each concentration and baseline. The mean, standard deviation and %CV were calculated for the predicted and bach concentration. Linear regression analysis of actual Tnl concentration versus expected TnI concentration resulted with an R = 0.997 and a slope of 1.019 with an offset of 0.279. The recovery of spiked Tnl antigen at the five concentrations was 0.82, 1.72, 3.65, 7.65, 15.78, and 27.51 ng/mL. Percent recovery of Tnl ranged from 95 to 115% with an average of 105%.

HOOK EFFECT: There is no high dose hook effect in the RAMP Troponin I Assay up to the highest level tested (500 ng/mL Tnl).

ANALYTICAL SENSITIVITY: The lower limit of detection (LLD) is defined as the analyte concentration corresponding to the mean (n=20) plus 2 standard deviations of the zero. The LLD is 0.03 ng/mL Tnl. Tnl levels in excess of 32 ng/mL are reported as greater than (>) 32 nq/mL.

ANALYTICAL SPECIFICITY: Potentially cross-reactive substances were evaluated by spiking different concentrations into blood. Skeletal Troponin I, Cardiac Troponin T and Cardiac Troponin C appear to have no significant cross-reactivity with the RAMP Troponin I Assay. HAMA, HAGA, HARA and RhF appear to have minimal cross-reactivity with the RAMP Troponin I Assay.

INTERFERENCE: Potentially interfering substances were evaluated by spiking different concentrations of potential interferents into EDTA blood with TnI added. Different blood samples were used for each potential interferent. Interference was evaluated by calculating the Tnl concentration of potential interferent-spiked blood, expressed as a percentage of the Tnl concentration of the unspiked (no potential interferent) blood sample. No evidence of cross-reactivity or interference was observed for hemoglobin, triglyceride, bilirubin, cholesterol, or heparin at levels of very high physiological concentrations, up to 1500 mg/dL, 3000mg/dL, 80 mg/dL, 500 mg/dL, and 66 IU/mL, respectively. No trend was observed in the Tnl predictions as the concentration of potential interferent was increased.

CLINICAL PERFORMANCE

EXPECTED VALUES

Whole blood samples from 180 healthy individuals (84 males and 96 females) were assayed for Troponin I by the three methods, RAMP, Triage, and Dimension. The lower assayed for Troponia 1 by the first normal range were defined as the 5" and 95" percentile values and are presented in Table 4-1. The RAMP Troponin I normal range distribution is presented in Figure 4-1.

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The normal range of the RAMP Troponin I Assay was found to be 0.00 ng/mL in the The normal fange of the Triage system reports values less than 0.2 as "< 0.2" and hemial popular the normal range of the Triage Troponin I Assay was found to be < 0.2 because of this the normal fange of d. This is very similar to the normal range described ngmlE in the normal popular of < 0.19 ng/mL [1]. Finally, the Dimension normal range was found to be 0.00 to 0.04 ng/mL in the normal population studied. This is very similar to the normal range described in the Dimension Troponin I package insert of 0.00 to 0.05 ng/mL [2]. The data is presented in Table 4-1.

	RAMPng/mL	Triageng/mL	Dimensionng/mL
Percentile
5th (LLN)	0.00	< 0.2	0.00
50th	0.00	< 0.2	0.00
90th	0.00	< 0.2	0.04
95th (ULN)	0.00	< 0.2	0.04
97.5th	0.025	< 0.2	0.04
Figure	11-1	11-2	11-3

Table 4-1: Percentile Ranking of Normal Individuals

Figure 4-1: RAMP Normal Range Distribution

Image /page/3/Figure/5 description: The image is a graph showing frequency and cumulative percentage of RAMP Troponin I (ng/mL). The x-axis represents RAMP Troponin I (ng/mL) values from 0.00 to >0.20, while the left y-axis represents frequency and ranges from 0 to 200. The right y-axis represents cumulative percentage and ranges from 0% to 100%. The graph includes a bar chart representing frequency and a line graph representing cumulative percentage.

PRECISION STUDY

One hundred and eighty-four (184) subjects were enrolled in the Precision Study. Of these, 55 were normal individuals (28 males and 27 females) and 129 were patients suspected of AMI based on the individual hospital criteria (76 males and 53 females). The samples were selected from those obtained during the Method Comparison Study. The samples were stored refrigerated for up to one day between analyses. The data were reviewed and one outlier was removed. Correlation for Troponin I Assay replicate Result 2 vs Result 1 for the RAMP Assay is presented in Table 4-2.

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Population	n	Slopey =	Intercept	R
Combined Populations	183	1.086	-0.153	0.989
Patients with suspect AMI	128	1.093	-0.246	0.988

Table 4-2: Precision of RAMP Troponin I Assay, Result 1 vs Result 2

METHOD COMPARISON

Three hundred and sixty-five (365) subjects were enrolled in the Method Comparison Study. Of these subjects, 180 were normal individuals (84 males and 96 females) and 185 were suspected of acute myocardial infarct (AMI) based on the individual hospital criteria (115 males and 70 females). EDTA and heparin whole blood samples were obtained for each of these subjects. All normal subjects were consented. Waste samples were used for the subjects suspected of AMI. An aliquot of the whole blood was taken for the RAMP Troponin I Assay and heparinized plasma was prepared for the Triage Troponin I Assay and the Dimension Troponin I Assay. The samples were stored refrigerated for up to one day between analyses for the rapid tests. Heparin samples were frozen and sent to a reference lab for the Dimension testing.

To accommodate the differing reportable ranges of the RAMP and Triage Troponin I Assays, and the Dimension Troponin I Assay, the data were winsorized, and then examined for outliers. The correlation data is presented for both the RAMP and Triage Troponin I Assays versus the Dimension Troponin I Assay in Table 4-3.

	Population	n	Slope (y=)	Intercept	R
RAMP Troponin I Assay	Combined Normal andSuspect AMI Subjects	364	0.456	0.011	0.988
	Suspect AMI Subjects	184	0.456	0.025	0.986
Triage Troponin I Assay	Combined Normal andSuspect AMI Subjects	365	0.718	-0.138	0.974
	Suspect AMI Subjects	185	0.729	-0.563	0.972

Table 4-3: Correlation of RAMP and Triage vs Dimension

8. Conclusion

The RAMP Troponin I Assay when utilized with the RAMP Reader is substantially equivalent to other assays currently in commercial distribution for the measurement of Thi.

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Image /page/5/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.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

MAY 1 7 2004

Mr. William J. Radvak President and CEO Response Biomedical Corp. 8081 Lougheed Hwy. Burnaby, British Columbia, Canada V5A 1W9

Re: K033745

Trade/Device Name: RAMP Troponin 1 Assay Regulation Number: 21 CFR 862.1215 Regulation Name: Creatine phosphokinase/creatine kinase or isoenzymes test system Regulatory Class: Class II Product Code: MMI Dated: March 15, 2004 Received: March 15, 2004

Dear Mr. Radvak:

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 r ou mays 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 (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 or any I with all the Act's requirements, including, but not limited to: registration and listing (21 CVR 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) I mis lotter will and wyour e 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, If you don't spotition 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). You may obtain other general information on your responsibilities under the Act from the I ou may 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,

Jean M. Cooper, US, DVM.

Jean M. Cooper, MS, D.V.M. Director Division of Chemistry and Toxicology Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known): K033745

RAMP Troponin I Assay Device Name:

The RAMP Troponin I Assay is a quantitative Indications For Use: immunochromatographic test indicated for use as an in vitro diagnostic product used with the RAMP Clinical Reader to measure cardiac troponin I levels in EDTA whole blood. Measurement of cardiac troponin I aids in the rapid diagnosis of acute myocardial infarction (AMI). The RAMP Troponin I Assay is intended to be used only to patient management for those prioritize suspected of AMI.

(Part 21 CFR 801 Subpart D)

$\frac{(VD)}{(D)}$ AND/OR

Prescription Use X (IVD) AND/OR Over-The-Counter Use N/A (21 CFR 807 Subpart C)

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

Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD)

Abuts Gils'
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Office of in Vitro Diagnostic Device Evaluation and Safety

510(k)

Page 1 of 1

Regulation Number and Section

§ 862.1215 Creatine phosphokinase/creatine kinase or isoenzymes test system.

(a)
Identification. A creatine phosphokinase/creatine kinase or isoenzymes test system is a device intended to measure the activity of the enzyme creatine phosphokinase or its isoenzymes (a group of enzymes with similar biological activity) in plasma and serum. Measurements of creatine phosphokinase and its isoenzymes are used in the diagnosis and treatment of myocardial infarction and muscle diseases such as progressive, Duchenne-type muscular dystrophy.(b)
Classification. Class II.