VITROS Immunodiagnostic Products hs Troponin I Reagent Pack

K252393 · Ortho-Clinical Diagnostics, Inc. · MMI · Oct 29, 2025 · Clinical Chemistry

Device Facts

• Record ID: K252393
• Device Name: VITROS Immunodiagnostic Products hs Troponin I Reagent Pack
• Applicant: Ortho-Clinical Diagnostics, Inc.
• Product Code: MMI · Clinical Chemistry
• Decision Date: Oct 29, 2025
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1215
• Device Class: Class 2

Intended Use

For in vitro diagnostic use only. For the quantitative measurement of cardiac troponin I (cTnl) in human plasma (heparin) using the VITROS 5600 Integrated System. Cardiac troponin I is used to aid in the diagnosis of myocardial infarction (MI).

Device Story

VITROS Immunodiagnostic Products hs Troponin I Reagent Pack is an in vitro diagnostic immunoassay for quantitative measurement of cardiac troponin I (cTnI) in lithium heparin plasma. Performed on the VITROS 5600 Integrated System; device uses streptavidin-conjugated antibody and HRP-labeled antibody conjugate to bind cTnI in patient samples. Antigen-antibody complex is captured on biotin-BSA coated wells; unbound components are removed via washing. Bound HRP conjugate is measured via luminescent luminol reaction; light signal is proportional to cTnI concentration. Used in emergency departments by clinical laboratory personnel. Results aid physicians in diagnosing myocardial infarction. Benefits include rapid, quantitative assessment of cardiac injury to support clinical decision-making for patients with suspected acute coronary syndrome.

Clinical Evidence

Prospective, multicenter, blinded study of 2145 patients (≥22 years) presenting with acute coronary syndrome symptoms. Adjudicated diagnosis (MI vs. non-MI) based on Fourth Universal Definition of MI. Sensitivity, specificity, NPV, and PPV calculated for 0-2h, 2-4h, 4-6h, and 6-11h intervals using sex-specific (9 ng/L female, 12 ng/L male) and overall (11 ng/L) cut-offs. Sensitivity ranged from 84.03% to 97.62% depending on interval and cut-off. False negative rates were 3.2% (female) and 8.0% (male) using sex-specific cut-offs.

Technological Characteristics

Quantitative immunoassay; biotin-BSA/streptavidin-mouse monoclonal anti-cTnI capture; HRP-mouse monoclonal anti-cTnI conjugate. Measuring range 2.25–30,000 ng/L. Instrument: VITROS 5600 Integrated System. Detection via luminescent luminol reaction. Traceable to internal standard. Complies with CLSI EP05-A3, EP17-A2, EP06, EP07, EP37, EP28-A3c, and ISO 17511:2021.

Indications for Use

Indicated for the quantitative measurement of cardiac troponin I (cTnI) in human heparin plasma to aid in the diagnosis of myocardial infarction (MI) in patients presenting with symptoms consistent with acute coronary syndrome.

Regulatory Classification

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.

Predicate Devices

• Vitros Troponin I ES Reagent Pack (K062838)

Submission Summary (Full Text)

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FDA U.S. FOOD & DRUG ADMINISTRATION

October 29, 2025

Ortho-Clinical Diagnostics, Inc.
Kristine Tkacs
Senior Manager, Regulatory Affairs
100 Indigo Creek Drive
Rochester, NY 14626

Re: K252393
Trade/Device Name: VITROS Immunodiagnostic Products hs Troponin I Reagent Pack
Regulation Number: 21 CFR 862.1215
Regulation Name: Creatine phosphokinase/creatine kinase or isoenzymes test system
Regulatory Class: Class II
Product Code: MMI
Dated: July 31, 2025
Received: July 31, 2025

Dear Kristine Tkacs:

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 (the 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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database available at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

U.S. Food & Drug Administration
10903 New Hampshire Avenue
Silver Spring, MD 20993
www.fda.gov

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K252393 - Kristine Tkacs
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Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).

Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30, Design controls; 21 CFR 820.90, Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review, the QS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).

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 Part 801 and Part 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reporting-combination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.

All medical devices, including Class I and unclassified devices and combination product device constituent parts are required to be in compliance with the final Unique Device Identification System rule ("UDI Rule"). The UDI Rule requires, among other things, that a device bear a unique device identifier (UDI) on its label and package (21 CFR 801.20(a)) unless an exception or alternative applies (21 CFR 801.20(b)) and that the dates on the device label be formatted in accordance with 21 CFR 801.18. The UDI Rule (21 CFR 830.300(a) and 830.320(b)) also requires that certain information be submitted to the Global Unique Device Identification Database (GUDID) (21 CFR Part 830 Subpart E). For additional information on these requirements, please see the UDI System webpage at https://www.fda.gov/medical-devices/device-advice-comprehensive-regulatory-assistance/unique-device-identification-system-udi-system.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-devices/medical-device-safety/medical-device-reporting-mdr-how-report-medical-device-problems.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medical-devices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-devices/device-advice-comprehensive-regulatory-

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K252393 - Kristine Tkacs
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assistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

Paula V. Caposino -S

Paula Caposino, Ph.D.
Deputy Director
Division of Chemistry and
Toxicology Devices
OHT7: Office of In Vitro Diagnostics
Office of Product Evaluation and Quality
Center for Devices and Radiological Health

Enclosure

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FORM FDA 3881 (8/23)
Page 1 of 1
PSC Publishing Services [phone]
EF

| DEPARTMENT OF HEALTH AND HUMAN SERVICES Food and Drug Administration Indications for Use | Form Approved: OMB No. 0910-0120 Expiration Date: 07/31/2026 See PRA Statement below. |
| --- | --- |
| 510(k) Number (if known) K252393 | |
| Device Name VITROS Immunodiagnostic Products hs Troponin I Reagent Pack | |
| Indications for Use (Describe) For In Vitro Diagnostic Use | |
| For the quantitative measurement of cardiac troponin I (cTnI) in human plasma (heparin) using the VITROS 5600 Integrated System. Cardiac troponin I is used to aid in the diagnosis of myocardial infarction (MI). | |
| Type of Use (Select one or both, as applicable) ☑ Prescription Use (Part 21 CFR 801 Subpart D) ☐ Over-The-Counter Use (21 CFR 801 Subpart C) | |
| CONTINUE ON A SEPARATE PAGE IF NEEDED. | |
| This section applies only to requirements of the Paperwork Reduction Act of 1995. *DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.* | |
| The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to: Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff [e-mail] | |
| "An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number." | |

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K252393 - Page 1 of 17

# 510(k) Summary

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: K252393

## Applicant

Ortho-Clinical Diagnostics, Inc.
100 Indigo Creek Drive
Rochester, NY 14626

Contact Person: Kristine Tkacs
Phone: [phone]

## Device Name

**Trade or Proprietary Names:**
VITROS® Immunodiagnostic Products hs Troponin I Reagent Pack

**Common Name:**
VITROS hs Troponin I Assay

**Classification:**
Creatine phosphokinase/creatine kinase or isoenzymes test system (862.1215), Class II

**Product Code:**
MMI

## Predicate Device

VITROS Immunodiagnostic Products hs Troponin I Reagent Pack referred to as the VITROS hs Troponin I assay (test) in the remainder of this document, is substantially equivalent to the VITROS Immunodiagnostic Products Troponin I ES Reagent Pack K062838.

## Device Description

The VITROS hs Troponin I assay is performed using the VITROS Immunodiagnostic Products hs Troponin I Reagent Pack and the VITROS Immunodiagnostic Products hs Troponin I Calibrators on the VITROS 5600 Integrated System. An immunometric immunoassay technique is used. Cardiac troponin I present in the sample reacts simultaneously with streptavidin-conjugated antibody (mouse monoclonal anti- cTnI), bound by biotin-BSA on the wells, and a horseradish peroxidase (HRP)-labeled antibody conjugate (mouse monoclonal anti-cTnI). The antigen-antibody complex is captured by the antibody on the wells. Unbound materials are removed by washing. The bound HRP conjugate is measured by a luminescent reaction. A reagent containing

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luminogenic substrates (a luminol derivative and a peracid salt) and an electron transfer agent is added to the wells. The HRP in the bound conjugate catalyzes the oxidation of the luminol derivative, producing light. The electron transfer agent (a substituted acetanilide) increases the level of light produced and prolongs its emission. The light signals are read by the system. The amount of HRP conjugate bound is directly proportional to the concentration of cardiac troponin I present.

## Device Intended Use

For in vitro diagnostic use only.

For the quantitative measurement of cardiac troponin I (cTnI) in human plasma (heparin) using the VITROS 5600 Integrated System.

Cardiac troponin I is used to aid in the diagnosis of myocardial infarction (MI).

## Comparison to Predicate Device

Table 1 provides a summary of the key features of the new device assessed against the predicate.

Table 1: Summary of Key Features of VITROS Immunodiagnostic Products hs Troponin I Reagent Pack compared to the Predicate Device

| Device Characteristic | Predicate Device
VITROS Immunodiagnostic Products
Troponin I ES Reagent Pack, K062838 | New Device
VITROS Immunodiagnostic Products
hs Troponin I Reagent Pack, K252393 |
| --- | --- | --- |
| Similarities | | |
| Indication for Use | For in vitro diagnostic use only.
For the quantitative measurement of cardiac troponin I (cTnI) in human plasma (heparin) using the VITROS 5600 Integrated System.
Cardiac troponin I is used to aid in the diagnosis of myocardial infarction (MI). | Same |
| Analyte | Cardiac Troponin I | Same |
| Antibody | Mouse Monoclonal | Same |
| Type of immunoassay | Sandwich immunoassay | Same |

K252393 - Page 2 of 17

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| Device Characteristic | Predicate Device VITROS Immunodiagnostic Products Troponin I ES Reagent Pack, K062838 | New Device VITROS Immunodiagnostic Products hs Troponin I Reagent Pack, K252393 |
| --- | --- | --- |
| Similarities | | |
| Detection technology | Chemiluminescence | Same |
| Traceability | Calibration of the VITROS Troponin I ES test is traceable to in-house reference calibrators, which have been value assigned to correlate to another commercially available test. | Same |
| Device Characteristic | Predicate Device VITROS Immunodiagnostic Products Troponin I ES Reagent Pack, K062838 | New Device VITROS Immunodiagnostic Products hs Troponin I Reagent Pack, K252393 |
| --- | --- | --- |
| Differences | | |
| Indication for Use | For the quantitative measurement of cardiac Troponin I (cTnI) in human serum (EDTA) Indicated for the risk stratification of patients with non-ST-segment elevation acute coronary syndromes with respect to relative risk of mortality, myocardial infarction (MI) or increased probability of ischemic events requiring urgent revascularization procedures | Not indicated for these uses |
| Sample Type | Serum and plasma (EDTA and lithium heparin) | Lithium heparin plasma |
| Measuring Range | 12-80,000 ng/L | 2.25–30,000 ng/L |
| Detection Limit | LoB: 0.007 ng/mL LoD: 0.012 ng/mL LoQ: 0.012 ng/mL | LoB: 0.19 ng/L LoD: 0.59 ng/L LoQ: 2.25 ng/L |
| Upper 99th percentile cut-off | 34 ng/L | Female: 9 ng/L Male: 12 ng/L Overall: 11 ng/L |

# Nonclinical performance

The following nonclinical tests were performed.

# Precision

Precision was evaluated consistent with CLSI document EP05-A3. Two replicates each of four

K252393 - Page 3 of 17

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human lithium heparin plasma patient pools and two controls were tested on two separate occasions per day on at least 20 different test days. The experiment was performed using three reagent lots on one VITROS 5600 Integrated System. The test was designed to have within-laboratory precision of ≤20% at or near the LoQ, ≤10% at the 99th percentile, and <7% between the 99th percentile and 30,000 ng/L. Representative performance data are shown below.

| VITROS System | Conc. Units - ng/L (pg/mL) | | | | | | | No. Observe. | No. Days |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | Mean hs Troponin I Conc. | Within-run* | | Within-calibration** | | Within-lab*** | | | |
| | | SD | CV (%) | SD | CV (%) | SD | CV (%) | | |
| 5600 | 5.48 | 0.141 | 2.7 | 0.254 | 4.9 | 0.422 | 7.4 | 80 | 20 |
| | 11.66 | 0.308 | 2.7 | 0.566 | 4.9 | 0.633 | 5.4 | 80 | 20 |
| | 65.27 | 1.143 | 1.8 | 1.907 | 3.0 | 2.602 | 3.9 | 80 | 20 |
| | 309.2 | 3.42 | 1.1 | 9.15 | 3.0 | 9.56 | 3.0 | 80 | 20 |
| | 14,090 | 262.7 | 1.9 | 415.8 | 3.0 | 764.8 | 5.4 | 80 | 20 |
| | 18,460 | 236.3 | 1.3 | 565.0 | 3.1 | 544.8 | 2.9 | 80 | 20 |

* Within-run (repeatability). Between Duplicate precision averaged over all runs.
** Within-calibration. Total precision with weighted components of within-run, between-run, and between-day variation.
*** Within-lab. A measure of the effect of recalibration on total precision, calculated within reagent lot, using data from at least 4 calibrations.

## Detection Limit

The Limit of Detection (LoD) for the VITROS hs Troponin I test was consistent with CLSI document EP17-A2. The observed LoD was determined to be 0.59 ng/L. The observed Limit of Quantitation (LoQ) at 20% CV was determined to be 0.59 ng/L, consistent with CLSI document EP17-A2. The claimed LoQ is set at 0.59 ng/L.

| LoD | LoQ (20% CV) |
| --- | --- |
| 0.59 ng/L | 0.59ng/L |

## Linearity

Linearity studies were performed in line with CLSI document EP06 2nd edition. The linearity of the VITROS hs Troponin I test was established following CLSI EP06-ED2. For cTnI, the measurement procedure shows linearity for the interval from 2.25 to 30,000 ng/L, with deviations from linearity within +/- 10%.

Two studies were conducted. The first study used test fluids comprised of human plasma (troponin-stripped/low protease/buffer matrix) and spanned the entire measuring interval using 11 levels with a minimum of 3 replicates for each level, the observed % deviation from linearity ranged from -4.2% to 2.3%. The second study used lithium heparin plasma patient samples and spanned the low end up to 4799 ng/L with 12 levels and a minimum of 5 replicates for each level, the observed % deviation from linearity ranged from -9.0% to 8.5%.

K252393 - Page 4 of 17

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Analytical Specificity

# Known Interferences

The VITROS hs Troponin I test was evaluated for interference in lithium heparin plasma samples with cTnI concentrations of approximately 10.00 ng/L and 350.0 ng/L, consistent with CLSI EP07 3rd edition and EP37 1st edition. The following compounds, when tested at the concentrations indicated, caused the biases shown.

| Substance | Interferent concentration | cTnI conc. | Effect when above the concentration limit | % Interference |
| --- | --- | --- | --- | --- |
| Bilirubin, unconjugated | 40 mg/dL | 11.2 ng/L | Decreased cTnI results | -10% |
| | 40 mg/dL | 330 ng/L | Decreased cTnI results | -6% |
| Cefoxitin Sodium | 523 mg/dL | 11.2 ng/L | Decreased cTnI results | -8% |
| Dextran | 600 mg/dL | 13.1 ng/L | Increased cTnI results | +8% |
| | 600 mg/dL | 385 ng/L | Increased cTnI results | +5% |
| Fibrinogen | 500 mg/dL | 8.8 ng/L | Decreased cTnI results | -3% |
| Hemoglobin | 750 mg/dL | 320 ng/L | Decreased cTnI results | -9% |
| Streptokinase* | 37,500 U/dL | 12.0 ng/L | Decreased cTnI results | -79% |
| | 37,500 U/dL | 390 ng/L | Decreased cTnI results | -65% |
| tPA (alteplase) | 0.6 mg/dL | 11.8 ng/L | Decreased cTnI results | -9% |
| | 0.3 mg/dL | 386 ng/L | Decreased cTnI results | -5% |
| Total protein** | 10.2 g/dL | 10.3 ng/L | Decreased cTnI results | -1% |
| | 10.1 g/dL | 390 ng/L | Increased cTnI results | +3% |

*Streptokinase is not commercially available in the United States.
**Test results compared to a control sample with total protein concentration of 7.3 g/dL.

Note: These results are representative. The degree of interference at concentrations other than those listed might not be predictable from these results. Other interfering substances may be encountered in the patient population.

# Other Limitations

- Dextran at therapeutic doses significantly interferes with this test. This test should not be used on patients taking Dextran. An alternate method not subject to Dextran interference should be used.
- Heterophile as well as human anti-animal antibodies (most common human anti-mouse antibodies or HAMA) in serum or plasma of certain individuals are known to cause interference with immunoassays.¹ The anti-animal antibodies may be present in blood samples from individuals regularly exposed to animals or who have received preparations

K252393 - Page 5 of 17

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of mouse monoclonal antibodies for diagnosis or therapy. Results inconsistent with clinical observations indicate the need for additional testing.

- Troponin autoantibodies have been reported to be present in approximately 10% to 20% of patients presenting to the emergency department (ED) and may lead to falsely low troponin assay results and delay in treatment of acute coronary syndrome.²,³ Therefore, a test result that is inconsistent with the clinical picture and patient history should be interpreted with caution.

## Substances that do not Interfere

The VITROS hs Troponin I test was evaluated for interference in lithium heparin plasma samples consistent with CLSI EP07 and EP37. Of the compounds tested, none was found to cause a bias of >10% with the test at the concentrations indicated at nominal cTnI concentrations of 10.00 ng/L and 350.0 ng/L.

| Compound | Concentration | |
| --- | --- | --- |
| Acetaminophen | 156 μg/mL | 1030 μmol/L |
| Acetylcysteine | 15 mg/dL | 919 μmol/L |
| Adrenaline (epinephrine) | 20 μg/dL | 1.09 μmol/L |
| Allopurinol | 6.0 mg/dL | 441 μmol/L |
| Alprazolam | 0.0258 mg/dL | 0.835 μmol/L |
| Ambroxol | 63 μg/dL | 1.52 μmol/L |
| Amlodipine besylate | 0.0104 mg/dL | 0.183 μmol/L |
| Amoxicillin | 5.40 mg/dL | 148 μmol/L |
| Ascorbic acid | 5.25 mg/dL | 298 μmol/L |
| Atorvastatin calcium | 0.162 mg/dL | 1.34 μmol/L |
| Benazepril HCl | 0.044 mg/dL | 0.954 μmol/L |
| Bilirubin, conjugated | 40 mg/dL | 475 μmol/L |
| Biotin | 3510 ng/mL | 14,300 nmol/L |
| Bivalirudin | 2.18 mg/dL | 10.0 μmol/L |
| Caffeine | 10.8 mg/dL | 556 μmol/L |
| Carvedilol | 43.2 μg/dL | 1.06 μmol/L |
| Ceftriaxone disodium hemi(heptahydrate) | 100 mg/dL | 1510 μmol/L |
| Cephalexin sodium | 13.4 mg/dL | 363 μmol/L |
| Cholesterol | 400 mg/dL | 10.3 mmol/L |
| Cinnarizine | 108 μg/dL | 2.93 μmol/L |
| Clopidogrel | 2.4 μg/dL | 0.057 μmol/L |
| Cocaine | 0.6 mg/dL | 17.7 μmol/L |
| Cotinine | 0.24 mg/dL | 4.89 μmol/L |
| Cyclosporin A | 0.18 mg/dL | 1.50 μmol/L |
| Digoxin | 0.0039 mg/dL | 0.0499 μmol/L |

² Park JY, Jaffe AS. Troponin autoantibodies: from assay interferent to mediator of cardiotoxicity. Clin Chem 2017; 63(1):30-32.
³ Nussinovitch U, Shoenfeld Y. Anti-troponin autoantibodies and the cardiovascular system. Heart 2010;96(19):1518-1524

K252393 - Page 6 of 17

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K252393 - Page 7 of 17
| Compound | Concentration | |
| --- | --- | --- |
| Diphenhydramine | 0.0774 mg/dL | 3.03 μmol/L |
| Dopamine | 0.0621 mg/dL | 4.06 μmol/L |
| Enalaprilat | 0.0819 mg/dL | 2.35 μmol/L |
| Enoxaparin (LMWH) | 360 U/dL | N/A |
| Eptifibatide | 1.44 mg/dL | 17.3 μmol/L |
| Erythromycin | 13.8 mg/dL | 188 μmol/L |
| Ethanol | 600 mg/dL | 130,000 μmol/L |
| Fondaparinux | 0.39 mg/dL | 2.26 μmol/L |
| Furosemide | 1.59 mg/dL | 48.1 μmol/L |
| HAMA (human anti-mouse antibodies) | 800 μg/L | 0.005 μmol/L |
| Heparin (Sodium), UFH | 330 U/dL | N/A |
| Ibuprofen | 21.9 mg/dL | 1060 μmol/L |
| Insulin | 3.12 μg/dL | 0.005 μmol/L |
| L-dopa (Levodopa) | 0.75 mg/dL | 38.0 μmol/L |
| Levothyroxine | 0.0429 mg/dL | 0.552 μmol/L |
| Lidocaine | 1.5 mg/dL | 64.0 μmol/L |
| Methylprednisolone | 0.783 mg/dL | 20.9 μmol/L |
| Metronidazole | 12.3 mg/dL | 719 μmol/L |
| Naproxen sodium | 39.3 mg/dL | 1560 μmol/L |
| Nifedipine | 0.0588 mg/dL | 1.70 μmol/L |
| Nitrofurantoin | 0.213 mg/dL | 8.94 μmol/L |
| Nitroglycerin (Nitrostat) | 1.2 μg/dL | 0.053 μmol/L |
| Omeprazole | 0.84 mg/dL | 24.3 μmol/L |
| Oxycodone | 0.0324 mg/dL | 1.03 μmol/L |
| Oxytetracycline | 1.2 mg/dL | 24.2 μmol/L |
| Phenytoin | 6.0 mg/dL | 238 μmol/L |
| Propranolol HCl | 0.115 mg/dL | 3.88 μmol/L |
| Pseudoephedrine | 0.330 mg/dL | 20.0 μmol/L |
| Quinidine | 1.5 mg/dL | 46.2 μmol/L |
| Rheumatoid Factor | 900 IU/mL | N/A |
| Rifampicin (Rifampin) | 4.8 mg/dL | 58.3 μmol/L |
| Rivaroxaban | 0.270 mg/dL | 6.19 μmol/L |
| Salicylic acid | 2.86 mg/dL | 207 μmol/L |
| Spironolactone | 0.0555 mg/dL | 1.33 μmol/L |
| Theophylline | 6.0 mg/dL | 333 μmol/L |
| Total Protein | 10.1 g/dL | 101 g/L |
| Triglycerides | 1500 mg/dL | 16.9 mmol/L |
| Vancomycin hydrochloride | 12.3 mg/dL | 82.8 μmol/L |
| Verapamil | 0.16 mg/dL | 3.51 μmol/L |
| Vorapaxar | 36 μg/dL | 0.731 μmol/L |
| Warfarin sodium | 8.0 mg/dL | 259 μmol/L |

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# Cross-Reactivity

The cross-reactivity of the VITROS hs Troponin I test was evaluated in lithium heparin plasma by adding the following substances to the respective plasma pools containing no cTnI.

| Cross-Reactant | Cross Reactant Concentration | | Mean Result of Control Pool | Mean Result of Cross-Reactant Pool | % Cross-Reactivity |
| --- | --- | --- | --- | --- | --- |
| | ng/mL | ng/L | ng/L (pg/mL) | ng/L (pg/mL) | |
| | | | | | |
| Actin (from Rabbit Muscle) | 1000 | 1000000 | * | * | * |
| Cardiac Troponin C (Recombinant) | 1000 | 1000000 | * | 11.01 | 0.0 |
| Cardiac Troponin T (Recombinant) | 1000 | 1000000 | * | * | * |
| CK-MB (Recombinant) | 1000 | 1000000 | * | * | * |
| Myoglobin (Recombinant) | 1000 | 1000000 | * | * | * |
| Myosin (Recombinant) | 1000 | 1000000 | * | * | * |
| Skeletal Troponin I | 1000 | 1000000 | * | * | * |
| Tropomyosin (from porcine muscle) | 1000 | 1000000 | * | * | * |

* Not Detectable. Concentration was below the measuring range of the test, 2.25 - 30,000 ng/L (pg/mL).

The cross-reactivity of the VITROS hs Troponin I test was evaluated in lithium heparin plasma by adding the following substances to the respective plasma pools containing cTnI at a concentration of approximately 10.00 ng/L.

| Cross-Reactant | Cross Reactant Concentration | | Mean Result of Control Pool | Mean Result of Cross-Reactant Pool | % Cross-Reactivity |
| --- | --- | --- | --- | --- | --- |
| | ng/mL | ng/L | ng/L (pg/mL) | ng/L (pg/mL) | |
| | | | | | |
| Actin (from Rabbit Muscle) | 1000 | 1000000 | 11.65 | 11.94 | 0.0 |
| Cardiac Troponin C (Recombinant) | 1000 | 1000000 | 11.65 | 26.70 | 0.0 |
| Cardiac Troponin T (Recombinant) | 1000 | 1000000 | 11.65 | 11.64 | 0.0 |
| CK-MB (Recombinant) | 1000 | 1000000 | 12.39 | 14.53 | 0.0 |
| Myoglobin (Recombinant) | 1000 | 1000000 | 12.39 | 11.73 | 0.0 |
| Myosin (Recombinant) | 1000 | 1000000 | 12.39 | 11.35 | 0.0 |
| Skeletal Troponin I | 1000 | 1000000 | 11.65 | 15.31 | 0.0 |
| Tropomyosin (from porcine muscle) | 1000 | 1000000 | 12.39 | 13.69 | 0.0 |

Cross-reactivity was expressed as the mean result obtained for the cross-reactant pool minus the mean result obtained for the control sample divided by the cross-reactant concentration in percentage terms.

$$
\% \text{ Cross-reactivity} = (\text{Mean Troponin I Result Cross-reactant Pool}) - (\text{Mean Troponin I Result Control Sample}) \times 100
$$

Concentration of Cross-reactant

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cTnC was spiked into one Lithium Heparin plasma pool with endogenous cTnI at approximately 10.00 ng/L. cTnC (Purified from Human heart tissue, 98% purity) was prepared as a concentrate before spiking into the sample pool to yield a cTnC concentration of 80,000 ng/L. An equal volume of the relevant solvent was added to each of the control pools for comparison. Interim dilutions were also prepared at 75%, 50% and 25% of the high pool, yielding concentrations of 60,000 ng/L, 40,000 ng/L and 20,000 ng/L.

Five replicates were collected for each fluid level. The percent bias between the test/interim dilutions and control samples was calculated: 100 * (Spike - Control Sample)/Control Sample.

| | VITROS hsTnI | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Cardiac Troponin C (ng/L) | Rep 1 (ng/L) | Rep 2 (ng/L) | Rep 3 (ng/L) | Rep 4 (ng/L) | Rep 5 (ng/L) | Mean (ng/L) | % Difference (Mean - Mean_Blank)/Mean_Blank |
| 0 | 10.04 | 10.63 | 10.24 | 10.63 | 10.29 | 10.37 | N/A |
| 20000 | 10.56 | 11.03 | 10.50 | 10.77 | 10.57 | 10.69 | 3.1% |
| 40000 | 11.95 | 12.30 | 11.39 | 11.65 | 11.34 | 11.73 | 13.1% |
| 60000 | 12.40 | 12.24 | 12.28 | 12.29 | 12.14 | 12.27 | 18.3% |
| 80000 | 11.93 | 11.89 | 11.85 | 11.89 | 11.76 | 11.87 | 14.5% |

The results of these tests demonstrate that the performance of the VITROS hs Troponin I assay is not affected by the presence of cTnC up to 20,000 ng/L.

## High Dose Hook

The VITROS hs Troponin I test has no high dose hook effect up to a concentration of 100,000 ng/L.

## Expected Values

The VITROS hs Troponin I 99th percentile URLs were established from lithium heparin plasma of nine hundred fifty-two (952) apparently healthy adults, including 486 female and 466 male subjects. The results included variation from reagent lots, systems, operators, and reagent age. The subjects ranged in age from 22 to 91 years old, with fifty-nine percent of the subjects ≥ 50 years of age.

Subjects were excluded if they met any of the following exclusion criteria:

- History of kidney disease, diabetes, heart disease, cancer, lung disease, thyroid disease, or stroke
- High blood pressure, cholesterol or triglycerides
- Muscle or skeletal injury or surgery in the last 3 months
- Current smoker
- Pregnancy
- Additional exclusion criteria:
- Hemoglobin A1c ≥ 6.5%
- NT-proBNP > 125 pg/mL for subjects < 75 years of age or > 450 pg/mL for subjects ≥ 75

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years of age

- eGFR < 60 mL/min/1.73m²

The overall observed 99th percentile URL from the 952 lithium heparin plasma samples is 11 ng/L (90% CI of 8.2 – 14.3 ng/L). The 99th percentile URL values and respective 90% Confidence Intervals (CI), determined for lithium heparin plasma (females, males, and overall) using the non-parametric statistical method, are shown in the table below.

| Sample Type | Gender | Number of Subjects | 99th Percentile URL ng/L (pg/mL) | 90% CI* ng/L (pg/mL) |
| --- | --- | --- | --- | --- |
| Lithium Heparin Plasma | Female | 486 | 9 | 3.9 - 17.5 |
| | Male | 466 | 12 | 8.8 - 20.9 |
| | Overall | 952 | 11 | 8.2 - 14.3 |

*CI = Confidence Interval

## High Sensitivity Determination

For a troponin test to be labelled as high sensitivity it must fulfil the two IFCC TF-CB established criteria:

- The test should have analytical imprecision (% CV) at the 99th percentile ≤ 10% and
- The test should measure troponin concentrations above the Limit of Detection (LoD) in ≥ 50% of healthy male and subjects ≥ 50% of healthy female subjects.

Data from a Limit of Quantitation (LoQ) study was used to create a best fit regression expressing the relationship between %CV and cTnI concentration in the clinically relevant range. The analytical imprecision around the 99th percentile URL concentrations was estimated using the regression analysis.

| Sample Type | Gender | 99th PercentileURL ng/L (pg/mL) | %CV Based on LoQ Imprecision Profile |
| --- | --- | --- | --- |
| Lithium Heparin Plasma | Female | 9 | 4.7 |
| | Male | 12 | 4.5 |
| | Overall | 11 | 4.5 |

Greater than 50% of the healthy male subjects and greater than 50% of the healthy female subjects used to establish the 99th percentile had cTnI results equal to or above the claimed LoD. The VITROS hs troponin I test meets the IFCC TF-CB criteria for a high sensitivity cardiac troponin test.⁴

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# Clinical performance

Results of this test should be used in conjunction with the patient's history, risk factors, clinical presentation, other diagnostic tests, such as electrocardiogram (ECG), and in accordance with the appropriate clinical guidelines to aid in the diagnosis of MI.

A multi-center prospective study was conducted to assess the diagnostic accuracy of the VITROS hs Troponin I test in lithium heparin plasma.

The study included 2145 evaluable subjects from patients presenting with symptoms consistent with acute coronary syndrome (ACS) at 24 Emergency Departments (ED) across the United States. Each subject's diagnosis was determined by an independent adjudication committee consisting of US board-certified cardiologists according to the Fourth Universal Definition of Myocardial Infarction – consensus guideline. The prevalence of adjudicated MI in the study was 8.16% (175/2145).

The diagnostic accuracy of the VITROS hs Troponin I test was determined by quantifying the area under the Receiver Operating Characteristic (ROC) curves (AUC) for each time bracket relative to the time of patient ED presentation, within and across genders. The AUC and the two-tailed 95% confidence intervals (CIs) are presented in the table below: AUC Analysis Results for the VITROS hs Troponin I Test.

AUC Analysis Results for the VITROS hs Troponin I Test

| Gender | Time Bracket | N | AUC | 95% Confidence Interval |
| --- | --- | --- | --- | --- |
| Female | 0-2 hours | 1042 | 0.947 | 0.911- 0.983 |
| | ≥2-4 hours | 858 | 0.979 | 0.966- 0.992 |
| | ≥4-6 hours | 508 | 0.963 | 0.930- 0.996 |
| | ≥6-11 hours | 541 | 0.976 | 0.962- 0.991 |
| Male | 0-2 hours | 1214 | 0.902 | 0.868- 0.937 |
| | ≥2-4 hours | 979 | 0.914 | 0.875- 0.952 |
| | ≥4-6 hours | 648 | 0.920 | 0.876- 0.963 |
| | ≥6-11 hours | 799 | 0.937 | 0.906- 0.969 |

Applications of Cardiac Bio-Markers of the International Federation of Clinical Chemistry and Laboratory Medicine. Clin Chem. 2018;64(4):645-655. doi:10.1373/clinchem.2017.277186.
5 Thygesen K, Alpert JS, Jaffe AS, Chaitman BR, Bax JJ, Morrow DA, et al. Fourth Universal Definition of Myocardial Infarction (2018). Circulation 2018; 138: e618-e651. https://doi.org/10.1161/CIR.000000000000617
6 Zweig MH, Campbell G, Receiver-Operating Characteristic (ROC) Plots: A Fundamental Evaluation Tool in Clinical Medicine. Clinical Chemistry, 1993; 39(4):561-577.
7 Bamber D, The Area Above the Ordinal Dominance Graph and the Area Below the Receiver Operating Characteristic Graph. Journal of Mathematical Psychology. 1975; 12:387-415.

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AUC Analysis Results for the VITROS hs Troponin I Test

| Gender | Time Bracket | N | AUC | 95% Confidence Interval |
| --- | --- | --- | --- | --- |
| All Subjects | 0-2 hours | 2256 | 0.923 | 0.898- 0.948 |
| | ≥2-4 hours | 1837 | 0.941 | 0.918- 0.965 |
| | ≥4-6 hours | 1156 | 0.935 | 0.905- 0.965 |
| | ≥6-11 hours | 1340 | 0.952 | 0.931- 0.972 |

Clinical performance was determined by the following analyses:

| VITROS hs Troponin I Test Result | Adjudicated Diagnosis | |
| --- | --- | --- |
| | MI | No MI |
| cTnI >99th percentile cutoff | A | B |
| cTnI ≤99th percentile cutoff | C | D |

- Sensitivity = 100xA/(A+C);
- Specificity = 100xD/(B+D);
- Negative predictive value (NPV), probability of no MI diagnosis in patients with cTnI ≤99th percentile cutoff = 100xD/(C+D);
- Positive predictive value (PPV), probability of MI diagnosis in patients with cTnI >99th percentile cutoff = 100xA/(A+B);
- Negative likelihood ratio (LR-), the ratio of the likelihood of false negative results to the likelihood of true negative results = (1-sensitivity)/specificity;
- Positive likelihood ratio (LR+), the ratio of the likelihood of true positive results to the likelihood of false positive results = sensitivity/(1-specificity).

Predictive value analysis is directly related to the prevalence of the disease in the intended use population. Likelihood ratios are not directly related to the prevalence of the disease in the intended use population and are applicable across populations with varying MI prevalence.

Data analyses were conducted with samples categorized into four time brackets related to patient ED presentation: 0-2 hours, ≥2-4 hours, ≥4-6 hours, and ≥6-11 hours.

The clinical performance of the VITROS hs Troponin I test versus adjudicated diagnosis was determined by time brackets after ED presentation for all subjects using the overall (11 ng/L) 99th percentile cutoff.

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All Subjects using 11 ng/L Cutoff (Overall 99th Percentile)

| Time Bracket | N | Sensitivity (%) (n/N) | 95% CI* (%) | Specificity (%) (n/N) | 95% CI* (%) | NPV (%) (n/N) | 95% CI* (%) | PPV (%) (n/N) | 95% CI* (%) |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| 0-2 hours | 2256 | 85.64 (155/181) | 79.78 - 90.00 | 86.60 (1797/2075) | 85.07 - 88.00 | 98.57 (1797/1823) | 97.92 - 99.02 | 35.80 (155/433) | 31.42 - 40.42 |
| ≥2-4 hours | 1837 | 90.00 (144/160) | 84.37 - 93.75 | 85.45 (1433/1677) | 83.68 - 87.06 | 98.90 (1433/1449) | 98.21 - 99.32 | 37.11 (144/388) | 32.45 - 42.02 |
| ≥4-6 hours | 1156 | 92.00 (115/125) | 85.90 - 95.60 | 81.57 (841/1031) | 79.09 - 83.82 | 98.82 (841/851) | 97.85 - 99.36 | 37.70 (115/305) | 32.45 - 43.27 |
| ≥6-11 hours | 1340 | 93.30 (167/179) | 88.65 - 96.12 | 78.55 (912/1161) | 76.10 - 80.82 | 98.70 (912/924) | 97.74 - 99.26 | 40.14 (167/416) | 35.54 - 44.92 |

* 95% Wilson Score Confidence Interval

| Time Bracket | N | LR- | 95% CI** | LR+ | 95% CI* |
| --- | --- | --- | --- | --- | --- |
| 0-2 hours | 2256 | 0.17 | 0.12 - 0.24 | 6.39 | 5.64 - 7.24 |
| ≥2-4 hours | 1837 | 0.12 | 0.07 - 0.19 | 6.19 | 5.45 - 7.02 |
| ≥4-6 hours | 1156 | 0.10 | 0.05 - 0.18 | 4.99 | 4.35 - 5.73 |
| ≥6-11 hours | 1340 | 0.09 | 0.05 - 0.15 | 4.35 | 3.87 - 4.89 |

* 95% Confidence Interval

The clinical performance of the VITROS hs Troponin I test versus adjudicated diagnosis was determined by time brackets after ED presentation for female subjects using the gender-specific (9 ng/L) and overall (11 ng/L) 99th percentile cutoffs. Using the female gender-specific (9 ng/L) 99th percentile cutoff instead of the overall (11 ng/L) 99th percentile cutoff may result in a higher proportion of positive test results for females that are not having an MI.

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Female Subjects Using Overall (11 ng/L) versus Gender-specific (9 ng/L) 99th Percentile Cutoffs

| Time Bracket | 99th Percentile Cutoff | Sensitivity (%) (n/N) | 95% CI* (%) | Specificity (%) (n/N) | 95% CI* (%) | NPV (%) (n/N) | 95% CI* (%) | PPV (%) (n/N) | 95% CI* (%) |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| 0-2 hours (N=1042) | Gender-specific | 88.71 (55/62) | 78.48 - 94.42 | 90.31 (885/980) | 88.29 - 92.00 | 99.22 (885/892) | 98.39 - 99.62 | 36.67 (55/150) | 29.38 - 44.62 |
| | Overall | 87.10 (54/62) | 76.55 - 93.31 | 91.63 (898/980) | 89.73 - 93.21 | 99.12 (898/906) | 98.27 - 99.55 | 39.71 (54/136) | 31.87 - 48.10 |
| ≥2-4 hours (N=858) | Gender-specific | 95.08 (58/61) | 86.51 - 98.31 | 88.58 (706/797) | 86.19 - 90.61 | 99.58 (706/709) | 98.76 - 99.86 | 38.93 (58/149) | 31.47 - 46.94 |
| | Overall | 91.80 (56/61) | 82.21 - 96.45 | 90.21 (719/797) | 87.95 - 92.09 | 99.31 (719/724) | 98.39 - 99.70 | 41.79 (56/134) | 33.78 - 50.26 |
| ≥4-6 hours (N=508) | Gender-specific | 97.62 (41/42) | 87.68 - 99.58 | 84.98 (396/466) | 81.45 - 87.94 | 99.75 (396/397) | 98.59 - 99.96 | 36.94 (41/111) | 28.54 - 46.21 |
| | Overall | 95.24 (40/42) | 84.21 - 98.68 | 86.48 (403/466) | 83.08 - 89.29 | 99.51 (403/405) | 98.22 - 99.86 | 38.83 (40/103) | 29.99 - 48.49 |
| ≥6-11 hours (N=541) | Gender-specific | 96.77 (60/62) | 88.98 - 99.11 | 84.76 (406/479) | 81.27 - 87.70 | 99.51 (406/408) | 98.23 - 99.87 | 45.11 (60/133) | 36.91 - 53.59 |
| | Overall | 93.55 (58/62) | 84.55 - 97.46 | 86.01 (412/479) | 82.62 - 88.83 | 99.04 (412/416) | 97.55 - 99.63 | 46.40 (58/125) | 37.90 - 55.12 |

*95% Wilson Score Confidence Interval

| Time Bracket | 99th Percentile Cutoff | LR- | 95% CI* | LR+ | 95% CI** |
| --- | --- | --- | --- | --- | --- |
| 0-2 hours (N=1042) | Gender-specific | 0.13 | 0.06 - 0.25 | 9.15 | 7.41 - 11.30 |
| | Overall | 0.14 | 0.07 - 0.27 | 10.41 | 8.28 - 13.08 |
| ≥2-4 hours (N=858) | Gender-specific | 0.06 | 0.02 - 0.17 | 8.33 | 6.81 - 10.19 |
| | Overall | 0.09 | 0.04 - 0.21 | 9.38 | 7.50 - 11.73 |
| ≥4-6 hours (N=508) | Gender-specific | 0.03 | 0.00 - 0.19 | 6.50 | 5.21 - 8.11 |
| | Overall | 0.06 | 0.01 - 0.21 | 7.04 | 5.54 - 8.95 |
| ≥6-11 hours (N=541) | Gender-specific | 0.04 | 0.01 - 0.15 | 6.35 | 5.12 - 7.88 |
| | Overall | 0.08 | 0.03 - 0.19 | 6.69 | 5.31 - 8.43 |

*95% Confidence Interval

The lower bound of the CI for PPV for female subjects using the gender-specific 99th percentile (9 ng/L) cutoff was as low as 28.54% (≥4-6 hours). Cardiac troponin test results should always be interpreted in conjunction with the patient's signs, symptoms, and other clinical information.

The lower bound of the CI for PPV for female subjects using the overall 99th percentile (11 ng/L) cutoff was as low as 29.99% (at ≥4-6 hours). Cardiac troponin test results should always be interpreted in conjunction with the patient's signs, symptoms, and other clinical information.

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The clinical performance of the VITROS hs Troponin I test versus adjudicated diagnosis was determined by time brackets after ED presentation for male subjects using the gender-specific (12 ng/L) and overall (11 ng/L) 99th percentile cutoffs. Using the male gender-specific (12 ng/L) 99th percentile cutoff instead of the overall (11 ng/L) 99th percentile cutoff may result in a higher proportion of negative test results for males that are having an MI.

## Male Subjects Using Overall (11 ng/L) versus Gender-specific (12 ng/L) 99th Percentile Cutoffs

| Time Bracket | 99th Percentile Cutoff | Sensitivity (%) (n/N) | 95% CI* (%) | Specificity (%) (n/N) | 95% CI* (%) | NPV (%) (n/N) | 95% CI* (%) | PPV (%) (n/N) | 95% CI* (%) |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| 0-2 hours (N=1214) | Gender-specific | 84.03 (100/119) | 76.40 - 89.53 | 83.11 (910/1095) | 80.77 - 85.21 | 97.95 (910/929) | 96.83 - 98.69 | 35.09 (100/285) | 29.78 - 40.79 |
| | Overall | 84.87 (101/119) | 77.35 - 90.21 | 82.10 (899/1095) | 79.72 - 84.26 | 98.04 (899/917) | 96.92 - 98.75 | 34.01 (101/297) | 28.85 - 39.57 |
| ≥2-4 hours (N=979) | Gender-specific | 88.89 (88/99) | 81.19 - 93.68 | 81.48 (717/880) | 78.78 - 83.91 | 98.49 (717/728) | 97.31 - 99.15 | 35.06 (88/251) | 29.42 - 41.15 |
| | Overall | 88.89 (88/99) | 81.19 - 93.68 | 81.14 (714/880) | 78.42 - 83.58 | 98.48 (714/725) | 97.30 - 99.15 | 34.65 (88/254) | 29.06 - 40.69 |
| ≥4-6 hours (N=648) | Gender-specific | 89.16 (74/83) | 80.66 - 94.19 | 78.41 (443/565) | 74.83 - 81.60 | 98.01 (443/452) | 96.26 - 98.95 | 37.76 (74/196) | 31.27 - 44.72 |
| | Overall | 90.36 (75/83) | 82.12 - 95.03 | 77.52 (438/565) | 73.90 - 80.77 | 98.21 (438/446) | 96.50 - 99.09 | 37.13 (75/202) | 30.76 - 43.97 |
| ≥6-11 hours (N=799) | Gender-specific | 93.16 (109/117) | 87.09 - 96.49 | 75.07 (512/682) | 71.69 - 78.17 | 98.46 (512/520) | 96.99 - 99.22 | 39.07 (109/279) | 33.53 - 44.90 |
| | Overall | 93.16 (109/117) | 87.09 - 96.49 | 73.31 (500/682) | 69.87 - 76.50 | 98.43 (500/508) | 96.92 - 99.20 | 37.46 (109/291) | 32.09 - 43.15 |

*95% Wilson Score Confidence Interval

| Time Bracket | 99th Percentile Cutoff | LR- | 95% CI** | LR+ | 95% CI** |
| --- | --- | --- | --- | --- | --- |
| 0-2 hours (N=1214) | Gender-specific | 0.19 | 0.13 - 0.29 | 4.97 | 4.27 - 5.80 |
| | Overall | 0.18 | 0.12 - 0.28 | 4.74 | 4.09 - 5.50 |
| ≥2-4 hours (N=979) | Gender-specific | 0.14 | 0.08 - 0.24 | 4.80 | 4.11 - 5.60 |
| | Overall | 0.14 | 0.08 - 0.24 | 4.71 | 4.04 - 5.50 |
| ≥4-6 hours (N=648) | Gender-specific | 0.14 | 0.07 - 0.26 | 4.13 | 3.47 - 4.91 |
| | Overall | 0.12 | 0.06 - 0.24 | 4.02 | 3.40 - 4.76 |
| ≥6-11 hours (N=799) | Gender-specific | 0.09 | 0.05 - 0.18 | 3.74 | 3.25 - 4.30 |
| | Overall | 0.09 | 0.05 - 0.18 | 3.49 | 3.05 - 3.99 |

**95% Confidence Interval

The lower bound of the CI for PPV for male subjects using the gender-specific 99th percentile (12 ng/L) cutoff was as low as 29.42% (≥2-4 hours). Cardiac troponin test results should always be interpreted in conjunction with the patient's signs, symptoms, and other clinical information.

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The lower bound of the CI for PPV for male subjects using the overall 99th percentile (11 ng/L) cutoff was as low as 28.85% (0-2 hours). Cardiac troponin test results should always be interpreted in conjunction with the patient's signs, symptoms, and other clinical information.

## Non-elevated VITROS hs Troponin I Test Results in Patients Adjudicated as Having MI (False Negatives)

Among the 62 female subjects adjudicated as having MI, there were 4 for whom none of the VITROS hs Troponin I test results exceeded the overall cutoff of 11 ng/L. When using the gender-specific cutoff of 9 ng/L, the number of false negatives for females was reduced from 4 to 2.

Among the 113 male subjects adjudicated as having MI, there were 8 for whom none of the VITROS hs Troponin I test results exceeded the overall cutoff of 11 ng/L. When using the gender-specific cutoff of 12 ng/L, the number of false negatives for males increased from 8 to 9.

Serial blood samples for subjects in the VITROS hs Troponin I study were drawn no more than 11 hours after ED presentation; however, clinical information of the study subjects, including standard of care (SOC) troponin test results, were collected for the subjects' entire hospital stay.

The standard of care (SOC) troponin tests were the tests used by each clinical study site and were reviewed by the adjudicators for the adjudicated diagnosis.

For 2 of the male false negative subjects, the earliest SOC troponin test results that exceeded the SOC troponin cutoff were from samples drawn more than 26 hours after the final VITROS hs Troponin I sample was collected. For 2 of the female false negative subjects, the earliest SOC troponin test results that exceeded the SOC cutoff were from samples drawn more than 17 hours after the final VITROS hs Troponin I sample was collected. The adjudicated MI diagnosis for some of these subjects could therefore have been based on SOC troponin test results from samples drawn beyond the VITROS hs Troponin I study period.

## Elevated VITROS hs Troponin I Test Results in Patients Adjudicated as Not Having MI (False Positives)

There are conditions other than MI that are known to cause acute or chronic myocardial injury and lead to elevated troponin values. The VITROS hs Troponin I clinical trial enrolled all subjects presenting to the emergency department with symptoms consistent with ACS. Some of these subjects had an acute or chronic condition other than MI. In the clinical trial, 15.79% (311/1970) of subjects without an MI diagnosis had at least one VITROS hs Troponin I test result above the overall 99th percentile cutoff on one or more serial draws. 91.96% (286/311) of these subjects were found to have decreased kidney function (eGFR < 60 mL/min/1.73m², Stage 3 chronic kidney disease or renal failure) or one or more of the following cardiac conditions: angina, atrial fibrillation, cardiomyopathy, coronary artery disease, heart failure or tachycardia

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(heart rate > 100 BPM).

## Conclusions

The nonclinical and clinical data presented in the submission provides a reasonable assurance that the VITROS hs Troponin I Reagent Pack is substantially equivalent to the cleared predicate device.

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