Predicate Devices

Elecsys Troponin T Gen 5 STAT Immunoassay (K162895)

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The document describes a standard immunoassay and analyzer with no mention of AI or ML in the device description, performance studies, or key metrics. The analysis relies on established calibration curves and analytical methods.

Therapeutic

No.
This device is an immunoassay for diagnosing myocardial infarction, which is a diagnostic tool, not a therapeutic one. Therapeutic devices are used for treating diseases or conditions.

Diagnostic

Yes

The intended use specifies that the immunoassay is "intended to aid in the diagnosis of myocardial infarction."

SaMD (Software as a Medical Device)

No

The device is an immunoassay that relies on chemical reactions and detection on a physical analyzer (cobas e immunoassay analyzers) to determine the concentration of cardiac troponin T. It is not solely software.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states "Immunoassay for the in vitro quantitative determination of cardiac troponin T (cTnT) in lithium heparin plasma." The term "in vitro" is the defining characteristic of an IVD.
Sample Type: The assay is performed on "lithium heparin plasma," which is a biological sample taken from the body and tested outside of the body.
Purpose: The immunoassay is intended to "aid in the diagnosis of myocardial infarction," which is a medical condition. IVDs are used to diagnose, monitor, or screen for diseases or conditions.
Device Description: The description details a laboratory-based immunoassay process using reagents and an analyzer, which is typical of IVD devices.

The information provided clearly aligns with the definition of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

Immunoassay for the in vitro quantitative determination of cardiac troponin T (cTnT) in lithium heparin plasma. The immunoassay is intended to aid in the diagnosis of myocardial infarction.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on cobas e immunoassay analyzers.

Product codes (comma separated list FDA assigned to the subject device)

MMI

Device Description

The Elecsys Troponin T Gen 5 STAT Immunoassay is a one-step sandwich immunoassay on the cobas e 601 and cobas e 801 analyzer. The assay uses streptavidin-coated microparticles, a biotinylated monoclonal anti-cardiac Troponin T-specific antibody, a monoclonal anti-cardiac Troponin T-specific antibody labeled with a ruthenium complex and electrochemiluminescence detection. Results are determined using a calibration curve that is generated specifically on each instrument by a 2-point calibration and a master curve (6-point calibration) provided with the reagent bar code.

Mentions image processing

Not Found

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

Not Found

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Not Found

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

Not Found

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Precision/Reproducibility

Repeatability and Intermediate Precision: Evaluated according to CLSI guideline EP5-A3.
- Sample Size: Six human samples (sample pools) and two levels of PreciControl Troponin.
- Data Source: Native Li-Heparin plasma samples (≤ 100 ng/L cTnT) and spiked samples (> 100 ng/L cTnT).
- Setting: One cobas e 801 analyzer at one internal site with three reagent lots over 21 days for Repeatability and Intermediate Precision. Two replicates of each human Li-Heparin plasma sample and two levels of PreciControl Troponin were tested in two runs per day for 21 days.
- Key Results:
  - Sample 1 (9.60 ng/L): Repeatability SD = 0.225 ng/L (CV=2.3%), Intermediate precision SD = 0.405 ng/L (CV=4.2%)
  - Sample 2 (15.8 ng/L): Repeatability SD = 0.270 ng/L (CV=1.7%), Intermediate precision SD = 0.556 ng/L (CV=3.5%)
  - Sample 3 (22.2 ng/L): Repeatability SD = 0.310 ng/L (CV=1.4%), Intermediate precision SD = 0.643 ng/L (CV=2.9%)
  - Sample 4 (164 ng/L): Repeatability SD = 2.96 ng/L (CV=1.8%), Intermediate precision SD = 5.15 ng/L (CV=3.1%)
  - Sample 5 (4877 ng/L): Repeatability SD = 97.9 ng/L (CV=2.0%), Intermediate precision SD = 153 ng/L (CV=3.1%)
  - Sample 6 (9808 ng/L): Repeatability SD = 172 ng/L (CV=1.7%), Intermediate precision SD = 328 ng/L (CV=3.3%)
  - Control 1 (26.6 ng/L): Repeatability SD = 0.370 ng/L (CV=1.4%), Intermediate precision SD = 0.866 ng/L (CV=3.3%)
  - Control 2 (1962 ng/L): Repeatability SD = 27.2 ng/L (CV=1.4%), Intermediate precision SD = 51.7 ng/L (CV=2.6%)
Reproducibility: Evaluated according to CLSI guideline EP5-A3.
- Sample Size: Five human Li-Heparin plasma samples and two levels of PreciControl Troponin.
- Setting: Three cobas e 801 analyzers at three different sites (one internal and two external sites) over 5 days.
- Key Results:
  - Sample 1 (Mean 8.74 ng/L): Reproducibility SD = 0.770 ng/L (CV = 8.8%)
  - Sample 2 (Mean 21.6 ng/L): Reproducibility SD = 0.820 ng/L (CV = 3.8%)
  - Sample 3 (Mean 171 ng/L): Reproducibility SD = 5.99 ng/L (CV = 3.5%)
  - Sample 4 (Mean 4806 ng/L): Reproducibility SD = 174 ng/L (CV = 3.6%)
  - Sample 5 (Mean 9490 ng/L): Reproducibility SD = 282 ng/L (CV = 3.0%)
  - PreciControl 1 (Mean 25.4 ng/L): Reproducibility SD = 1.19 ng/L (CV = 4.7%)
  - PreciControl 2 (Mean 1923 ng/L): Reproducibility SD = 71.8 ng/L (CV = 3.7%)

Method Comparisons

Study Type: Comparison of the Elecsys Troponin T Gen 5 immunoassay on the cobas e 801 (Y-axis) with the predicate device (Elecsys Troponin T Gen 5 STAT Immunoassay) on the cobas e 601 (X-axis).
Sample Size: One set of 299 samples, with at least 264 samples detected within the measuring range.
Key Results:
- Passing-Bablok: Slope = 0.993 (95% LCL=0.979, 95% UCL=1.01), Y-intercept = 1.34 ng/L (95% LCL=1.07, 95%UCL=1.53), Absolute Bias at cutoff 14 ng/L = 1.24, Absolute Bias at cutoff 19 ng/L = 1.21, Absolute Bias at cutoff 22 ng/L = 1.19, Correlation Coefficient = 0.976 (Kendall's tau)
- Weighted Deming: Slope = 0.995 (95% LCL=0.987, 95% UCL=1.00), Y-intercept = 1.26 ng/L (95% LCL=1.10, 95%UCL=1.41), Absolute Bias at cutoff 14 ng/L = 1.19, Absolute Bias at cutoff 19 ng/L = 1.17, Absolute Bias at cutoff 22 ng/L = 1.16, Correlation Coefficient = 1.00 (Pearson's r)

Limit of Blank (LoB)

Study type: Determination of LoB according to CLSI EP17-A2.
Sample size: 60 measurements from analyte-free Li-Heparin plasma sample.
Key Results: All lots met the predetermined acceptance criterion of ≤ 2.5 ng/L. LoB claim in labeling will be set to 2.5 ng/L.

Limit of Detection (LoD)

Study type: Determination of LoD according to CLSI EP17-A2.
Sample size: 60 determinations per reagent lot (two-fold determination, six runs over at least three days for five low-analyte Li-Heparin plasma samples).
Key Results: All lots met the predetermined acceptance criterion of ≤ 3 ng/L. LoD claim in labeling will be set to 3 ng/L.

Limit of Quantitation (LoQ)

Study type: Determination of LoQ according to CLSI Guideline EP17-A2.
Sample size: Ten native Li-Heparin plasma pools and two controls.
Key Results: All lots met the predetermined acceptance criterion of 6 ng/L. LoQ claim in labeling will be set to 6 ng/L.

Linearity/Assay Reportable Range

Study type: Linearity study according to CLSI guideline EP6-A.
Sample size: Three high analyte Li-Heparin plasma samples diluted with three low analyte Li-Heparin plasma samples yielding at least sixteen concentrations.
Key Results: The linear range is from 6 to 13766 ng/L. The deviation from linearity is within specifications.

High Dose Hook Effect

Study type: Assessment of high dose hook effect.
Key Results: No High dose Hook effect was seen up to 111326 ng/L.

Human Anti-Mouse Antibodies (HAMA)

Study type: Assessment of HAMA interference.
Sample size: Two different samples with low levels of Troponin T spiked with 644 ug/L HAMA.
Key Results: Specifications were met for the two native Li-Heparin plasma samples. A maximum of 10% deviation from the expected concentration was seen for a HAMA concentration of 644 ug/L (80%).

Endogenous Interference Studies

Study type: Evaluation of endogenous substances for potential interference.
Interferents tested: Intralipid, biotin, bilirubin, hemoglobin, rheumatoid factors, cholesterol and human serum albumin.
Sample size: Three native Li-Heparin plasma samples (low, medium, high concentration of Troponin T) used for dilution series.
Key Results: No interference was seen from the potential interferents at the following levels: Biotin up to 1200 ng/mL, Intralipid up to 2000 mg/dL, Bilirubin up to 66.0 mg/dL, Hemoglobin up to 200 mg/dL, Rheumatic Factor up to 1200 IU/mL, Human Serum Albumin up to 7.00 g/dL, Cholesterol up to 310 mg/dL.

Cross-Reactivity

Study type: Evaluation for potential cross-reactivity.
Cross-reactants tested: Skeletal muscle TnT (30 ng/mL), Skeletal muscle TnI (100 ng/mL), Cardiac TnI (25 ng/mL), Human TnC (100 ng/mL).
Key Results: Specifications were met for all substances tested. No interference seen up to the spiked concentrations.

Exogenous Interference - Drugs

Study type: Evaluation of drugs for potential interference.
Sample size: Seventeen common pharmaceutical compounds and 22 cardiac drugs spiked into Li-Heparin plasma samples.
Key Results: Specifications were met for all exogenous interfering substances tested.

Clinical Data -- APACE Study

Study type: International, multicenter prospective trial evaluating diagnostic sensitivity and specificity for aid in diagnosis of MI.
Sample size: 1074 subjects with 3023 available samples, 188 adjudicated diagnoses of MI.
Data Source: Patients presenting to emergency department with symptoms of chest pain and angina pectoris, with peak symptoms within the last 12 hours (onset of symptoms reported ranged from 0 to 72 hours). Exclusion criteria: kidney failure requiring dialysis and age less than 18 years. Diagnosis of MI by independent adjudication committee.
Key Results (cobas e 801, single 99th percentile cutoff 19 ng/L, all sexes):
- BCT 1.5-2.5-3.5h: Sens = 91.8% (95% CI: 81.9-97.3), Spec = 82.4% (95% CI: 77.5-86.6), PPV = 52.3% (95% CI: 42.5-62.1), NPV = 98.0% (95% CI: 95.3-99.3)
Second Multicenter Study (cobas e 601, single 99th percentile cutoff 19 ng/L, all sexes):
- Sample Size: 1679 subjects enrolled, 173 adjudicated MIs. 1675 subjects evaluated on cobas e 601 analyzer.
- Exclusion criteria: MI within last 3 months, surgery/hospitalization within last 3 months, revascularization/PCI within last 3 months, established acute non-cardiac primary illness, subjects transferred from another hospital or facility.
- Key Results (females, single 99th percentile cutoff 19 ng/L):
  - Baseline: Sens = 82.5% (95% CI: 70.9-90.9), Spec = 91.9% (95% CI: 89.7-93.8), PPV = 47.7% (95% CI: 38.1-57.5), NPV = 98.3% (95% CI: 97.0-99.2)
  - 3 hours: Sens = 91.8% (95% CI: 80.4-97.7), Spec = 90.2% (95% CI: 87.6-92.4), PPV = 42.1% (95% CI: 32.6-52.0), NPV = 99.3% (95% CI: 98.2-99.8)
  - 6-9 hours: Sens = 91.3% (95% CI: 79.2-97.6), Spec = 90.8% (95% CI: 87.9-93.2), PPV = 48.3% (95% CI: 37.4-59.2), NPV = 99.1% (95% CI: 97.7-99.8)
  - 12-24 hours: Sens = 87.2% (95% CI: 72.6-95.7), Spec = 85.3% (95% CI: 81.2-88.8), PPV = 39.1% (95% CI: 28.8-50.1), NPV = 98.4% (95% CI: 96.3-99.5)
- Key Results (males, single 99th percentile cutoff 19 ng/L):
  - Baseline: Sens = 88.1% (95% CI: 80.2-93.7), Spec = 84.1% (95% CI: 81.2-86.7), PPV = 43.4% (95% CI: 36.5-50.5), NPV = 98.1% (95% CI: 96.7-99.0)
  - 3 hours: Sens = 95.6% (95% CI: 89.1-98.8), Spec = 83.0% (95% CI: 79.9-85.8), PPV = 44.4% (95% CI: 37.3-51.6), NPV = 99.3% (95% CI: 98.1-99.8)
  - 6-9 hours: Sens = 96.7% (95% CI: 90.8-99.3), Spec = 80.2% (95% CI: 76.5-83.5), PPV = 45.9% (95% CI: 38.7-53.2), NPV = 99.3% (95% CI: 98.0-99.9)
  - 12-24 hours: Sens = 94.4% (95% CI: 86.4-98.5), Spec = 76.3% (95% CI: 71.8-80.4), PPV = 41.7% (95% CI: 34.1-49.7), NPV = 98.7% (95% CI: 96.7-99.6)

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

See "Summary of Performance Studies" section for detailed metrics including Sensitivity, Specificity, Positive Predictive Value (PPV), and Negative Predictive Value (NPV) with 95% Confidence Intervals.

Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.

Elecsys Troponin T Gen 5 STAT Immunoassay (K162895)

Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).

Not Found

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.

Summary

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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: the Department of Health & Human Services logo on the left and the FDA text logo on the right. The text logo has the FDA acronym in a blue square, followed by the words "U.S. FOOD & DRUG ADMINISTRATION" in blue.

September 21, 2021

Roche Diagnostics Jane Phillips, Ph.D. Senior Regulatory Program Manager 9115 Hague Road Indianapolis, IN 46250

Re: K201441

Trade/Device Name: Elecsys Troponin T Gen 5 Regulation Number: 21 CFR 862.1215 Regulation Name: Creatine Phosphokinase/Creatine Kinase Or Isoenzymes Test System Regulatory Class: Class II Product Code: MMI Dated: December 2, 2020 Received: December 3, 2020

Dear Jane Phillips:

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. 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 located 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.

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

1

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 of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-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 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 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-device-safety/medical-device-reportingmdr-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/medicaldevices/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-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Marianela Perez-Torres, Ph.D. Deputy Director Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Ouality Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K201441

Device Name Elecsys Troponin T Gen 5

Indications for Use (Describe)

Immunoassay for the in vitro quantitative determination of cardiac troponin T (cTnT) in lithium heparin plasma. The immunoassay is intended to aid in the diagnosis of myocardial infarction.

The electrochemiluminescence immunoassay "ECLIA" is intended for use on cobas e immunoassay analyzers.

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)

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510(k) Summary of Safety and Effectiveness

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

510(k) Number	K201441
Manufacturer	Roche Diagnostics
9115 Hague Road
Indianapolis, IN 46250
Contact	Jane Ellen Phillips, PhD, Senior Regulatory Program
Manager
9115 Hague Road, Building B
jane.phillips@roche.com
Phone: 317-521-3338
Fax; 317-521-2324
Date	September 15th, 2021
Device Name	Proprietary name:	Elecsys Troponin T Gen 5
	Common name:	Troponin T Gen 5

Device Classification Class II

| Panel | Product
Code | Classification Name | Regulation
Citation |
|--------------------|-----------------|----------------------------------------------------------------------------|------------------------|
| Clinical Chemistry | MMI | Creatine
phosphokinase/creatine
kinase or isoenzymes test
system. | 862.1215 |

Substantial	The results presented in this 510(k) premarket notification
Equivalence	demonstrate that the Elecsys Troponin T Gen 5 Immunoassay is
	substantially equivalent to the predicate Elecsys Troponin T Gen
	5 STAT Immunoassay (K162895).

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| Substantial Equivalence Comparison | | Predicate Device:
Elecsys Troponin T Gen 5 STAT Immunoassay (K162895) |
|-----------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------|
| Feature | Candidate Device:
Elecsys Troponin T Gen 5 | |
| | General Immunoassay Features | |
| Intended Use/
Indications for
Use | Immunoassay for the in vitro quantitative
determination of cardiac troponin T (cTnT)
in lithium heparin plasma. The
immunoassay is intended to aid in the
diagnosis of myocardial infarction.

Intended Use of the Device:

Immunoassay for the in vitro quantitative determination of cardiac troponin T (cTnT) in lithium heparin plasma. The immunoassay is intended to aid in the diagnosis of myocardial infarction.

The electrochemluminescence immunoassay "ECLIA" is intended for use on the cobas e immunoassay analyzers.

Device Description

The Elecsys Troponin T Gen 5 STAT Immunoassay is a one-step sandwich immunoassay on the cobas e 601 and cobas e 801 analyzer. The assay uses streptavidin-coated microparticles, a biotinylated monoclonal anti-cardiac Troponin T-specific antibody, a monoclonal anti-cardiac

5

Troponin T-specific antibody labeled with a ruthenium complex and electrochemiluminescence detection. Results are determined using a calibration curve that is generated specifically on each instrument by a 2-point calibration and a master curve (6-point calibration) provided with the reagent bar code.

Changes to Reagent Composition

The following changes were implemented to the reagent to block the potential interference of biotin with the Elecsys Troponin T Gen 5 assay. Briefly, we took a two-step approach by adding an antibody to bind free biotin in the sample, and changing the linker on the biotinylated capture antibodv.

For the neutralization of free biotin in Li-Heparin plasma. Roche developed an antibody, which binds to free biotin. The antibodies are specific for free biotin and do not bind to or interact with the biotin-linker conjugates.

In the updated biotinylated antibody conjugate, the distance between biotin and the reactive MEA group is elongated by a PEG spacer, which provides the coupled biotin more flexibility for the interaction with the Streptavidin matrix. No change is made to the antibody (immunologically reactive component) which still recognizes the same epitope.

Summary of Analytical Performance

Precision/Reproducibility

Repeatability and Intermediate Precision

Precision measurements were conducted to evaluate repeatability (within-run precision) and the intermediate precision (within-laboratory precision) according to the CLSI guideline EP5-A3.

The precision of the Elecsys Troponin T Gen 5 assay was evaluated on one cobas e 801 analyzer at one internal site with three reagent lots over 21 days. Six human samples (sample pools) were run in randomized order on the analyzer. Native Li-Heparin plasma samples (≤ 100 ng/L cTnT) as well as spiked samples (> 100 ng/L cTnT) were used. Two replicates of each human Li-Heparin plasma sample and two levels of PreciControl Troponin were tested in two runs per day for 21 days. The analysis did not differentiate between runs conducted on the same day. therefore, the precision estimates reported below are for 21 days, 1 run/day, and 4 replicates. Calibration was performed once on day 1 according to the method sheet. No gel separator tubes were used in this experiment. Results from one representative lot of the three lots tested are described in the table below. Within laboratory precision includes within-run and between-day variability.

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| Sample | Mean
[ng/L] | Repeatability
(within-part precision) | | Intermediate precision
(within-lab/total) | |
|-----------|----------------|------------------------------------------|-----------|----------------------------------------------|-----------|
| | | SD
[ng/L] | CV
[%] | SD
[ng/L] | CV
[%] |
| Sample 1 | 9.60 | 0.225 | 2.3 | 0.405 | 4.2 |
| Sample 2 | 15.8 | 0.270 | 1.7 | 0.556 | 3.5 |
| Sample 3 | 22.2 | 0.310 | 1.4 | 0.643 | 2.9 |
| Sample 4 | 164 | 2.96 | 1.8 | 5.15 | 3.1 |
| Sample 5 | 4877 | 97.9 | 2.0 | 153 | 3.1 |
| Sample 6 | 9808 | 172 | 1.7 | 328 | 3.3 |
| Control 1 | 26.6 | 0.370 | 1.4 | 0.866 | 3.3 |
| Control 2 | 1962 | 27.2 | 1.4 | 51.7 | 2.6 |

Table 1.1: Results for Precision with the Reagent Lot 344524 according to the 21d measuring model

Reproducibility

Measurements were conducted to evaluate reproducibility according to the CLSI guideline EPS-A3.

A reproducibility panel consisting of five human Li-Heparin plasma samples and two levels of PreciControl Troponin was measured in five-fold determinations for 5 days on three cobas e 801 analyzers at three different sites (one internal and two external sites). The five native or spiked Li-Heparin plasma samples used were generated from sample pools. Samples ≤ 100 ng/L were native. For the higher concentrations, samples were spiked.

One cobas e 801 with a unique serial number per site was used with one reagent lot spanning one calibration cycle as recommended in the Method Sheet. Calibration was performed once on day one according to the method sheet. Repeatability, intermediate precision and reproducibility were calculated according to CLSI EP05-A3. Reproducibility includes repeatability, between day, and between site variance components.

| Sample Material | Mean
[ng/L] | Repeatability
(Within-run Precision) | | Intermediate Precision | |
|-----------------|----------------|-----------------------------------------|--------|------------------------|--------|
| | | SD [ng/L] | CV [%] | SD [ng/L] | CV [%] |
| Site 1 | | | | | |
| Sample 1 | 9.03 | 0.297 | 3.3 | 0.297 | 3.3 |
| Sample 2 | 22.0 | 0.323 | 1.5 | 0.350 | 1.6 |
| Sample 3 | 174 | 5.47 | 3.2 | 5.47 | 3.2 |

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| Sample Material | Mean
[ng/L] | Repeatability
(Within-run Precision) | |-----------------|----- | | | Sample 4 | 4890 | Sample 5 | 9540 | PreciControl 1 | 25.8 | PreciControl 2 | 1960 | Site 2 | | Sample 1 | 7.95 | Sample 2 | 20.9 | Sample 3 | 169 | Sample 4 | 4670 | Sample 5 | 9310 | PreciControl 1 | 24.3 | PreciControl 2 | 1860 | Site 3 | | Sample 1 | 9.24 | Sample 2 | 21.8 | Sample 3 | 170 | Sample 4 | 4860 | Sample 5 | 9620 | PreciControl 1 | 26.1 | PreciControl 2 | 1950 | Intermediate Precision | | |
-----------|-----------------------------------------|--------|------------------------|--------|--|
| SD [ng/L] | CV [%] | SD [ng/L] | CV [%] | |
| 44.2 | 0.9 | 62.2 | 1.3 | |
| 165 | 1.7 | 165 | 1.7 | |
| 0.455 | 1.8 | 0.469 | 1.8 | |
| 28.7 | 1.5 | 34.0 | 1.7 | |
| | | | | |
| 0.263 | 3.3 | 0.323 | 4.1 | |
| 0.418 | 2.0 | 0.418 | 2.0 | |
| 4.46 | 2.6 | 6.64 | 3.9 | |
| 94.5 | 2.0 | 143 | 3.1 | |
| 192 | 2.1 | 192 | 2.1 | |
| 0.594 | 2.5 | 0.868 | 3.6 | |
| 37.3 | 2.0 | 60.7 | 3.3 | |
| | | | | |
| 0.281 | 3.0 | 0.444 | 4.8 | |
| 0.376 | 1.7 | 0.832 | 3.8 | |
| 3.28 | 1.9 | 5.61 | 3.3 | |
| 67.6 | 1.4 | 174 | 3.6 | |
| 103 | 1.1 | 355 | 3.7 | |
| 0.378 | 1.5 | 0.877 | 3.4 | |
| 24.0 | 1.2 | 58.0 | 3.0 | |

Table 1.3: Results for Reproducibility

		Reproducibility
Sample Material	Mean
[ng/L]	SD
estimate
[ng/L]	SD
[ng/L]
95%
UCL	CV
estimate
(%)	CV (%)
95%
UCL
Sample 1	8.74	0.770	2.80	8.8	32.0
Sample 2	21.6	0.820	1.85	3.8	8.6
Sample 3	171	5.99	7.74	3.5	4.5

8

Sample Material	Mean [ng/L]	Reproducibility
		SD estimate [ng/L]	SD [ng/L] 95% UCL	CV estimate (%)	CV (%) 95% UCL
Sample 4	4806	174	342	3.6	7.1
Sample 5	9490	282	441	3.0	4.7
PreciControl 1	25.4	1.19	3.02	4.7	11.9
PreciControl 2	1923	71.8	154	3.7	8.0

Method Comparisons

Excellent comparison of the assay to the predicate device Elecsys Troponin T Gen 5 STAT is needed in order to ensure a transfer of the Roche clinical data from the existing and cleared assay to the biotin update.

To this end, method comparison studies were performed between one lot of the predicate device on the cobas e 601 and three lots of the Troponin T Gen 5 assay on the cobas e 801.

A method comparison (MC) study was performed to compare the Elecsys Troponin T Gen 5 immunoassay on the cobas e 801 (Y-axis) with the predicate device (X-axis) on the cobas e 601.

One set of samples was tested with one lot (predicate device, one internal site) and three different lots with the Elecsys Troponin T Gen 5 on the cobas e 801 (one internal site) as follows:

One set of 299 samples was tested with each reagent; at least 264 samples were detected within the measuring range. One lot of predicate device was taken as the reference value used for comparison to the values generated with each of the three lots of Elecsys Troponin T Gen 5 on the cobas e 801.

Table 1.4: Regression Analyses (n=264) Representative Method Comparison

	Passing-Bablok	Weighted Deming
Slope	0.993	0.995
95% LCL	0.979	0.987
95% UCL	1.01	1.00
Y-intercept [ng/L]	1.34	1.26
95% LCL	1.07	1.10
95%UCL	1.53	1.41
Absolute Bias at cutoff 14
[ng/L]	1.24	1.19
95% LCL	1.09	1.09
95%UCL

9

	Passing-Bablok	Weighted Deming
95% UCL	1.32	1.30
Absolute Bias at cutoff 19
[ng/L]	1.21	1.17
95% LCL	1.02	1.06
95% UCL	1.30	1.28
Absolute Bias at cutoff 22
[ng/L]	1.19	1.16
95% LCL	0.978	1.03
95% UCL	1.31	1.27
Correlation Coefficient	0.976 (Kendall's tau)	1.00 (Pearson's r)

Limit of Blank (LoB)

LoB of the Elecsy Troponin T Gen 5 assay on the cobas e 801 was determined according to CLSI EP17-A2.

Limit of Blank determines the highest observed measurement values for samples free of analyte. The Limit of Blank was determined as the 95th percentile of measurements of blank samples.

The distribution of values for the analyte-free Li-Heparin plasma sample was determined with three reagent lots on one cobas e 801 analyzer with six run over at least three days. The sample was measured in ten-fold determinations in each run.

In summary, 60 measurements were collected for the determination of LoB. The data were evaluated according to EP 17-A2, chapter 5.3.3.1 as the linear interpolation of the 57th and 58th ranked observation. The sample used was an analyte free native lithium-heparin sample.

Conclusion:

All lots met the predetermined acceptance criterion of ≤ 2.5 ng/L. The LoB claim in the labeling will be set to 2.5 ng/L.

Limit of Detection (LoD) (CLSI EP17-A2)

LoD of the Elecsys Troponin T Gen 5 assay on the cobas e 801 was determined according to CLSI EP17-A2. The LoD determines the lower limit for samples with analyte. The LoD was determined as the lowest amount of analyte in a sample that can be detected with a 95% probability.

10

Methods

The distribution of values for five low-analyte Li-Heparin plasma samples were determined using three reagent lots on one cobas e 801 instrument.

Five low-analyte Li-Heparin plasma samples were measured in two-fold determination, six runs over at least three days. In total. 60 determinations per reagent lot were performed.

In addition, we did testing for LoD on 3 different instruments by running samples on two additional e 801 instruments. Pooled estimate of the precision (SD total) of the five samples was calculated. The LoD was established according to the following EP17-A2 calculation:

LoD = LoB + 1.653 x SD total (of low analyte samples)

Conclusion: All lots met the predetermined acceptance criterion of ≤ 3 ng/L. The LoD claim in the labeling will be set to 3 ng/L.

Limit of Quantitation (CLSI EP17-A2)

The LoQ of the Elecsys Troponin T Gen 5 assay was determined on the cobas e 801 analyzer according to CLSI Guideline EP17-A2.

LoQ determines the lowest amount of analyte that can be quantitatively determined with stated accuracy and stated experimental conditions. The LoQ was determined as the lowest concentration of analyte which can be quantified with a CV (intermediate precision) of no more than 20%.

Methods

Functional sensitivity has been used as a detection capability performance attribute for the TnT Gen 5 Assay. It represents the measurand concentration associated with a desired within-lab (intermediate) precision, based upon a precision profile experiment in the low-end region of the measuring interval. This performance attribute, however, simply represents a limiting for the limit of quantitation (LoQ) in which the acceptable accuracy goal is based solely upon a precision requirement. This is suggested by CLSI EP17-A2 for troponin.

The performance goal was defined as intermediate precision equal to 20% CV.

Ten native Li-Heparin plasma pools were prepared across the low-end of the measuring range of the assay. In addition, two controls were included. Data were collected using three lots over 21 days with two runs per day. Estimates of the mean and within-lab precision were calculated for each sample for each reagent lot.

Total CV is based on the total variance, calculated as the sum of the variance components from day, run and within run (21 * 2 * 2 = 84 measurements). The square root from the total variance was divided by the grand mean times 100 is the result for the total CV (%). The calculation of

11

the variance components is based on a strict hierarchical model with random factors according to CLSI EP5-A3.

The functional relationship between CV and concentration is modeled according the suggestion in EP17 with:

%CV = A*conc B

To simplify the fit of the data, the CV and the mean are log-transformed. "Log" here refers to the natural logarithm. After log-transformation, a linear regression using least squares can be performed and the model looks like this:

log(%CV) = Ã + B * log(concentration)

A = log(A) With

Results are visually assessed and the goodness of fit of the data is evaluated.

The concentration where a 20% CV is achieved is calculated by:

log(20)-7 Concentration = exp (10) B

Conclusion:

All lots met the predetermined acceptance criterion of 6 ng/L. The LoQ claim in the labeling will be set to 6 ng/L.

Linearity/Assay Reportable Range

The linearity study was conducted to demonstrate the measurements across the claimed measuring range for each parameter to prove linearity. The study was performed according to CLSI guideline EP6-A.

Three high analyte Li-Heparin plasma samples from pools spiked with recombinant Troponin T were diluted with three low analyte Li-Heparin plasma samples from native pools. At least sixteen concentrations (15 dilutions) throughout the measuring range were prepared.

Samples were assayed in 3-fold determinations within a single run using one lot. Data from one representative sample is shown below

In the first step, a linearity check was performed with a first order (linear) regression analysis and then with higher order models (quadratic and cubic).

Data were analyzed using linear, quadratic and cubic order least square regression analysis according to CLSI protocol EP6-A.

12

y = a + b1 * x (first order polynomial or linear fit)

y = a + b1 * x + b2 * x2 (second order polynomial fit [quadratic])

y = a + b1 * x + b2 * x2 + b3 * x3 (third order polynomial fit [cubic])

Parameters of regression line

Intercept 2.94 ng/L, slope 0.966

The linear range is from 6 to 13766 ng/L

The coefficient b1 in the first order model is significant at the 5% level and no improvement of the goodness-of-fit for higher order models are observed. Therefore, it is used for the calculation of deviation from linearity.

Conclusions

The deviation from linearity is within specifications.

High Dose Hook Effect

Methods

The high dose hook effect of the Elecsys Troponin T Gen 5 assay was assessed on the cobas e 801 immunoassay analyzer.

Two low analyte Li-Heparin plasma samples from single donors were spiked with recombinant Troponin T to achieve high Troponin T concentrations. For each sample, a dilution series was performed using human Li-Heparin plasma, which contained no Troponin T.

The hook concentration reported corresponds to the analyte concentration with a signal corresponding to at least 10% above the highest master calibrator.

Conclusion:

No High dose Hook effect was seen up to 111326 ng/L

Human Anti-Mouse Antibodies (HAMA)

Methods

The effect of the presence of human anti-mouse-antibodies on the Troponin T Gen 5 assay was assessed on the cobas e 801 analyzer. Two different samples with low levels of Troponin T were spiked with potential interfering HAMA and tested in duplicate. The HAMA interferents used

13

for the interference testing have been quantified externally with a commercial assay and vielded a HAMA concentration of 644 ug/L.

Conclusion:

Specifications were met for the two native Li-Heparin plasma samples. A maximum of 10% deviation from the expected concentration was seen for a HAMA concentration of 644 uz/L (80%). In the labeling the testing of the specified HAMA concentration of 322 ug/L is reported.

Endogenous Interference Studies

The purpose of this study was to evaluate endogenous substances for potential interference with the parameters measured on the cobas e 801.

Methods

The effect on quantitation of analyte in the presence of endogenous interfering substances using the Elecsys Troponin T Gen 5 assay was determined on the cobas e 801 immunoassay analyzer for the following seven interfering substances: Intralipid, biotin, bilirubin, hemoglobin, rheumatoid factors, cholesterol and human serum albumin.

Three native Li-Heparin plasma samples (one low, one medium and one high concentration of Troponin T) were used to prepare dilution series. The series were tested with one lot.

Since biotin is of particular interest, we tested this kind of potential interference with three lots.

The following samples were used in this study:

Low sample: Native Li-Heparin plasma pool with analyte concentration of ~20 ng/L near the cut-off

Medium sample: Li-Heparin plasma pool of native samples with analyte concentration of ~100 ng/L in the slightly elevated part of the measuring range

High sample: Li-Heparin plasma pool of native samples with analyte concentration of ~900 ng/L in the elevated part of the measuring range

One part of each sample pool (low, medium, high) is spiked with the interfering endogenous substance and is used as "interference pool". Another part of the same sample pool is spiked with the same volume of the solvent of the interfering endogenous substance (without interfering substance) and is used as the related "dilution pool".

A series of nine dilution steps was prepared by mixing the interference pools and the related dilution pools.

Each sample was measured in 3-fold determination. For each interferent concentration level, the recovery compared to the "dilution pool" (without interfering substance) was calculated. For 3fold determinations, recovery calculation is based on the mean value.

Biotin concentrations of the samples were:50, 100, 500, 1000, 1250, 1500, 1750, 2000, 2500, 3600 ng/mL

14

Biotin Results

% Bias for samples containing various concentrations of biotin
Samples TnT
concentration (ng/L)	Biotin concentration (ng/mL)	50	100	500	1000	1250
18.9	-0.5	-1.3	2.0	-1.4	-2.5
108	0.3	1.8	1.7	-0.5	-2.2
870	-0.4	1.1	2.0	-0.1	-2.6

% Bias for samples containing various concentrations of biotin
Samples TnT
concentration (ng/L)	Biotin concentration (ng/mL)
	1500	1750	2000	2500	3600
18.9	-6.4	-12.2	-18.6	-39.7	-96.2
108	-6.9	-13.4	-20.9	-40.6	-93.2
870	-7.5	-12.7	-20.4	-40.6	-92.9

Note that patients with compromised renal function may have higher concentrations of biotin in their circulation.

Some studies have shown that serum concentrations of biotin can reach up to 355 ng/mL within the first hour after biotin ingestion for subjects consuming supplements of 20 mg of biotin per dayand up to 1160 ng/mL for subjects after a single dose of 300 mg biotin.

In addition, the following commonly used pharmaceuticals and cardiac specific drugs were tested (using cTnT concentrations of approximately 20 ng/L and 9000 ng/L). No interference with the assay was found.

Conclusions

No interference was seen from the potential interferents at the levels tabulated below.

15

Interferent Tested	No interference seen up to
Biotin	1200 ng/mL
Intralipid® (Lipemia)	2000 mg/dL
Bilirubin	66.0 mg/dL
Hemoglobin	200 mg/dL
Rheumatic Factor	1200 IU/mL
Human Serum Albumin	7.00 g/dL
Cholesterol	310 mg/dL

Table 1.5: Summary of Results for all Tested Interferents

Cross-Reactivity

This study was conducted to evaluate the Elecsys Troponin T Gen 5 assay on the cobas e 801 for potential cross-reactivity.

Methods

The analytical specificity of the Troponin T Gen 5 assay was assessed on the cobas e 801 analyzer using three concentrations of troponin in Li-Heparin plasma samples spiked with the potential cross-reacting compounds listed below:

Table 1.6: Cross-Reactants Tested

Interfering Substance	Spiked Concentration
Skeletal muscle TnT	30 ng/mL
Skeletal muscle TnI	100 ng/mL
Cardiac TnI	25 ng/mL
Human TnC	100 ng/mL

Description of samples used in cross-reactivity study: The spiked and non-spiked samples were tested in three-fold determination on the cobas e 801 analyzer.

Conclusion:

The specifications were met for all substances tested. See the following results summarized for the highest interference level tested and the resulting claim in the method sheet.

16

Interfering Substance	No interference seen up to	Method Sheet claim
Skeletal muscle TnT	30 ng/mL	30 ng/mL
Skeletal muscle TnI	100 ng/mL	100 ng/mL
Cardiac TnI	12.5 ng/mL	12.5 ng/mL
Human TnC	100 ng/mL	100 ng/mL

Table 1.7: Summarv of Cross-Reactivity Study

Exogenous Interference - Drugs

The purpose of this study was to evaluate drugs for potential interference with the Elecsys Troponin T Gen 5 assay on the cobas e 801.

Methods

Seventeen common pharmaceutical compounds were spiked into Li-Heparin plasma samples.

Native Li-Heparin plasma samples were used as the low sample was spiked with recombinant Troponin T.

The analyte concentrations were approximately 20 ng/L and 9000 ng/L.

In addition. 22 cardiac drugs were assessed by spiking into human Li-Heparin plasma samples. The analyte concentrations were approximately 19 ng/L and 8200 ng/L.

Li-Heparin plasma samples were divided into aliquots and spiked with the potential interferents. The reference sample without interferent was spiked with the respective amount of solvent only.

Testing was performed in 3-fold determination with one reagent lot in one run on two cobas e 801 analyzer.

Recovery was calculated based on the mean values of the 3-fold determinations.

Conclusion:

The specifications were met for all exogenous interfering substances tested.

Calibration and Traceability

This method has been standardized against the Elecsys Troponin T STAT assay (Mat# 04660307, 4th generation). This in turn was originally standardized against the Enzymun Test Troponin T (CARDIAC T) method.

17

Clinical Data -- APACE Study

Diagnostic sensitivity and specificity

APACE Trial: The Advantageous Predictors of Acute Coronary Syndromes Evaluation (APACE) study is an international, multicenter prospective trial of acute chest pain patients that is currently continuing enrollment. (ClinicalTrials.gov number NCT00470587). The sites enrolled all patients who presented to the emergency department with symptoms of chest pain and angina pectoris. Peak symptoms had to have occurred within the last 12 hours (onset of symptoms reported ranged from 0 to 72 hours). The only exclusion criterion were kidney failure that required dialysis and age less than 18 years. Diagnosis of MI was done through an independent adjudication committee which included cardiologists. This included 60 day followup information on each subject. In the case of a disagreement, a third independent cardiologist was used as the tie breaker. The subjects were diagnosed with acute MI by using the diagnostic criteria described in the ACC/ESC/AHA guidelines including ECG changes, symptoms characteristic for ischemia and elevations of cardiac troponin. 1074 subjects were enrolled with 3023 available samples. There were 188 adjudicated diagnoses of MI.

The clinical performance (clinical sensitivity, clinical specificity, positive predictive value [PPV] and negative predictive value [NPV]) of the Elecsys Troponin T Gen 5 STAT assay in the diagnosis of MI in this trial is shown below using a single 99th percentile cutoff 19 ng/L for all patients:

Performance data at all three cutoffs are presented below.

	Diagnosis
	MI	Non-MI
cTnT positive	A	B
cTnT negative	C	D

18

| Clinical performance of single 99th percentile cutoff (19 ng/L) for aid in diagnosis of AMI in

both sexes
Instrument	BCTa)	MDTb)	Nc)	Sensd)
%
95 % CIe)	Specf)
%
95 % CI	PPVg)
%
95 % CI	NPVh)
%
95 % CI
cobas e 801
analyzer	1.5-
2.5-
3.5	3.96	351	91.8 (81.9-
97.3)	82.4 (77.5-
86.6)	52.3 (42.5-
62.1)	98.0 (95.3-
99.3)

a) BCT = Blood collection time in hours since presentation

b) MDT = Mean draw time in hours

c) N = number of draws

d) Sensitivity = 100xA/(A+C)

e) CI = confidence interval

f) Specificity = 100xD/(B+D)

g) Positive predictive value = 100xA/(A+B)

h) Negative predictive value = 100xD/(D+C)

Clinical performance of single 99th percentile cutoff (19 ng/L) for aid in diagnosis of AMI in
women
Instrument	BCTa)	MDTb)	Nc)	Sensd)
%
95 % CIe)	Specf)
%
95 % CI	PPVg)
%
95 % CI	NPVh)
%
95 % CI
cobas e 801
analyzer	1.5-
2.5-
3.5	4.01	113	86.7 (59.5-
98.3)	86.7 (78.4-
92.7)	50.0 (29.9-
70.1)	97.7 (91.9-
99.7)

Clinical performance of single 99th percentile cutoff (19 ng/L) for aid in diagnosis of AMI in
men
Instrument	BCTa)	MDTb)	Nc)	Sensd)
%
95 % CIe)	Specf)
%
95 % CI	PPVg)
%
95 % CI	NPVh)
%
95 % CI
cobas e 801
analyzer	1.5-
2.5-
3.5	3.93	238	93.5 (82.1-
98.6)	80.2 (73.9-
85.6)	53.1 (41.7-
64.3)	98.1 (94.5-
99.6)

| Clinical performance of sex-specific 99th percentile cutoff

(14 ng/L) for aid in diagnosis of AMI in females
Instrument	BCTa)	MDTb)	Nc)	Sensd)
%
95 % CIe)	Specf)
%
95 % CI	PPVg)
%
95% CI	NPVh)
%
95% CI
cobas e 801
analyzer	1.5-
2.5-
3.5	4.01	113	93.3 (68.1-
99.8)	74.5 (64.7-
82.8)	35.9 (21.2-
52.8)	98.6 (92.7-
100.0)

| Clinical performance of sex-specific 99th percentile cutoff (22 ng/L) for aid in diagnosis of

AMI in males
Instrument	BCTa)	MDTb)	Nc)	Sensd)
%
95 % Cle)	Spec®
%
95 % CI	PPVg)
%
95 % CI	NPVh)
%
95 % CI
cobas e 801
analyzer	1.5-
2.5-
3.5	3.93	238	93.5 (82.1-
98.6)	83.3 (77.3-
88.3)	57.3 (45.4-
68.7)	98.2 (94.7-
99.6)

At least one of the following criteria should be met: symptoms of ischemia, ECG changes (ST and/or Q wave), left bundle branch block, imaging evidence of viable myocardium loss, wall motion abnormality or intracoronary thrombus to clarify the origin of myocardial injury.

In a second multicenter study, a total of 1679 subjects presenting emergently with chest pain were enrolled. The trial excluded chest pain subjects with an MI within the last 3 months,

21

subjects with surgery or hospitalization within the last 3 months, subjects with revascularization or percutaneous coronary intervention (PCI) within the last 3 months, subjects with an established acute non-cardiac primary illness and subjects transferred from another hospital or facility. These excluded subjects could be expected to have elevated troponin concentrations that would likely reflect cardiac comorbidities besides MI, and vield positive results; therefore specificity estimates and the positive values of this trial may be overestimated. Within this population, there were 173 adjudicated Mls. 1675 subjects were evaluated on the cobas e 601 analyzer. Final diagnoses were determined by an independent adjudication committee which included cardiologists and emergency medicine physicians using the universal guidelines.

The clinical performance of the Elecsys Troponin T Gen 5 STAT assay in the diagnosis of MI in this trial is shown below using a single 99th percentile cutoff (19 ng/L) for all patients and sexspecific cutoffs. The data are presented as timed draws (minutes) since presentation:

cobas e 601 analyzer
	MI	non-MI	Total
N	173	1502	1675
%	10.3	89.7	100

The agreement between Elecsys Troponin T Gen 5 and clinical diagnosis is shown in the tables below:

	Diagnosis
	MI	Non-MI
cTnT positive	A	B
cTnT negative	C	D

22

| Clinical performance of single 99th percentile cutoff (19 ng/L) for aid in diagnosis

of AMI in females
Instrument	BCTa)	Nb)	Sensc) %
95 % CId)	Spece) %
95 % CI	PPVf) %
95 % CI	NPVg) %
95 % CI
cobas e 601
analyzer	Baseline	771	82.5 (70.9-
90.9)	91.9 (89.7-
93.8)	47.7 (38.1-
57.5)	98.3 (97.0-
99.2)
	3 hours	682	91.8 (80.4-
97.7)	90.2 (87.6-
92.4)	42.1 (32.6-
52.0)	99.3 (98.2-
99.8)
	6-9 hours	536	91.3 (79.2-
97.6)	90.8 (87.9-
93.2)	48.3 (37.4-
59.2)	99.1 (97.7-
99.8)
	12-24
hours	399	87.2 (72.6-
95.7)	85.3 (81.2-
88.8)	39.1 (28.8-
50.1)	98.4 (96.3-
99.5)

a) BCT = Blood collection time in hours since presentation

b) N = number of draws
c) Sensitivity = 100xA/(A+C)

d) CI = confidence interval

e) Specificity = 100xD/(B+D)

f) Positive predictive value = 100xA/(A+B)
g) Negative predictive value = 100xD/(D+C)

| Clinical performance of single 99th percentile cutoff (19 ng/L) for aid in diagnosis

of AMI in males
Instrument	BCTa)	Nb)	Sensc) %
95 % CId)	Spece) %
95 % CI	PPVf) %
95 % CI	NPVg) %
95 % CI
cobas e 601
analyzer	Baseline	829	88.1 (80.2-
93.7)	84.1 (81.2-
86.7)	43.4 (36.5-
50.5)	98.1 (96.7-
99.0)
	3 hours	733	95.6 (89.1-
98.8)	83.0 (79.9-
85.8)	44.4 (37.3-
51.6)	99.3 (98.1-
99.8)

23

| Clinical performance of single 99th percentile cutoff (19 ng/L) for aid in diagnosis

of AMI in males
Instrument	BCTa)	Nb)	Sensc)
%
95 % CId)	Spece) %
95 % CI	PPVf)
%
95 % CI	NPVg)
%
95 % CI
	6-9 hours	622	96.7 (90.8-
99.3)	80.2 (76.5-
83.5)	45.9 (38.7-
53.2)	99.3 (98.0-
99.9)
	12-24
hours	473	94.4 (86.4-
98.5)	76.3 (71.8-
80.4)	41.7 (34.1-
49.7)	98.7 (96.7-
99.6)

| Clinical performance of sex-specific 99th percentile cutoff (14 ng/L) for aid in

diagnosis of AMI in females
Instrument	BCTa)	Nb)	Sensc)
%
95 % CId)	Spece) %
95 % CI	PPVf)
%
95 % CI	NPVg)
%
95 % CI
	Baseline	771	85.7
(74.6-
93.3)	88.1
(85.5-
90.4)	39.1
(30.9-
47.8)	98.6
(97.3-
99.3)
cobas e 601 analyzer	3 hours	682	91.8
(80.4-
97.7)	86.9
(84.0-
89.4)	35.2
(26.9-
44.1)	99.3
(98.2-
99.8)
	6-9 hours	536	91.3
(79.2-
97.6)	86.5
(83.2-
89.4)	38.9
(29.7-
48.7)	99.1
(97.6-
99.7)
	12-24
hours	399	92.3
(79.1-
98.4)	81.4
(77.0-
85.3)	35.0
(25.8-
45.0)	99.0
(97.1-
99.8)

24

| Clinical performance of sex-specific 99th percentile cutoff (22 ng/L) for aid in

diagnosis of AMI in males
Instrument	BCTa)	Nb)	Sensc)
%
95 % CId)	Spece) %
95 % CI	PPVf)
%
95 % CI	NPVg)
%
95 % CI
	Baseline	829	85.1
(76.7-
91.4)	87.2
(84.6-
89.6)	48.0
(40.5-
55.6)	97.7
(96.2-
98.7)
	3 hours	733	95.6
(89.1-
98.8)	86.3
(83.4-
88.9)	49.7
(42.1-
57.4)	99.3
(98.2-
99.8)
cobas e 601 analyzer	6-9 hours	622	93.5
(86.3-
97.6)	82.3
(78.7-
85.4)	47.8
(40.3-
55.3)	98.6
(97.1-
99.5)
	12-24
hours	473	94.4
(86.4-
98.5)	80.0
(75.8-
83.9)	45.9
(37.7-
54.3)	98.8
(96.9-
99.7)

Conclusion

The Elecsys Troponin T Gen 5 on the cobas e 801 is substantially equivalent to the FDA cleared Elecsys Troponin T Gen 5 STAT assay (K162895).