Alere NT-proBNP for Alinity i Reagent Kit

K241176 · Axis-Shield Diagnostics, Ltd. · NBC · Jan 16, 2025 · Clinical Chemistry

Device Facts

• Record ID: K241176
• Device Name: Alere NT-proBNP for Alinity i Reagent Kit
• Applicant: Axis-Shield Diagnostics, Ltd.
• Product Code: NBC · Clinical Chemistry
• Decision Date: Jan 16, 2025
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1117
• Device Class: Class 2

Intended Use

The Alere NT-proBNP for Alinity i assay is a chemiluminescent microparticle immunoassay (CMIA) used for the in vitro quantitative determination of N-terminal pro B-type natriuretic peptide (NT-proBNP) in human serum and plasma on the Alinity i system. In the emergency department, measurements of NT-proBNP are used as an aid in the diagnosis of heart failure (HF) in patients with clinical suspicion of new onset or worsening HF.

Device Story

The Alere NT-proBNP for Alinity i is an automated, two-step chemiluminescent microparticle immunoassay (CMIA) for quantitative determination of NT-proBNP in human serum and plasma. The device uses anti-NT-proBNP coated paramagnetic microparticles and acridinium-labeled conjugate. The system measures the resulting chemiluminescent reaction as relative light units (RLU), which are directly proportional to NT-proBNP concentration. Used in emergency departments, the device provides results that clinicians interpret alongside patient history and clinical examination to aid in diagnosing heart failure. The device helps differentiate between heart failure and other causes of dyspnea, potentially improving diagnostic accuracy and patient management in acute settings.

Clinical Evidence

Prospective multi-center study of 2127 ED subjects (18+ years) with suspected heart failure. Adjudication by 10 board-certified cardiologists. Performance evaluated using age-dependent positive cutoffs and age-independent negative cutoff. Results provided for all subjects, sex, age groups, and comorbidities (e.g., eGFR, BMI, history of HF). Sensitivity/specificity metrics derived from likelihood ratios and posttest probabilities. Study established clinical utility for aiding HF diagnosis.

Technological Characteristics

CMIA technology; reagents include anti-NT-proBNP (sheep, monoclonal) coated microparticles and anti-NT-proBNP (mouse, monoclonal) acridinium-labeled conjugate. Measuring interval: 15.8 to 35,000 pg/mL (extended to 350,000 pg/mL via dilution). Automated system (Alinity i). Standards referenced: CLSI EP05-A3, EP06, EP07, EP12, EP17-A2, EP28-A3c, EP34, EP35.

Indications for Use

Indicated for use in emergency departments as an aid in the diagnosis of heart failure (HF) in patients (18+ years) with clinical suspicion of new onset or worsening HF. Not indicated for patients with renal insufficiency requiring dialysis or known eGFR < 15.0 mL/min/1.73 m².

Regulatory Classification

Identification

The B-type natriuretic peptide (BNP) test system is an in vitro diagnostic device intended to measure BNP in whole blood and plasma. Measurements of BNP are used as an aid in the diagnosis of patients with congestive heart failure.

Special Controls

*Classification.* Class II (special controls). The special control is “Class II Special Control Guidance Document for B-Type Natriuretic Peptide Premarket Notifications; Final Guidance for Industry and FDA Reviewers.”

Predicate Devices

• Roche Elecsys proBNP II (K092649)

Submission Summary (Full Text)

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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo in blue, with the words "U.S. FOOD & DRUG" stacked on top of the word "ADMINISTRATION".

January 16, 2025

Axis-Shield Diagnostics Limited Claire Dora Associate Director, Regulatory Affairs 17 Luna Place, The Technology Park Dundee. Scotland DD 1XA United Kingdom

Re: K241176

Trade/Device Name: Alere NT-proBNP for Alinity i Reagent Kit Regulation Number: 21 CFR 862.1117 Regulation Name: B-Type Natriuretic Peptide Test System Regulatory Class: Class II Product Code: NBC Dated: December 11, 2024 Received: December 11, 2024

Dear Claire Dora:

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.

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"

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(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 OS 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 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-safetyreporting-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 Rue"). 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-device-advicecomprehensive-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-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

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For comprehensive regulatory information about mediation-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 ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

# Paula V. Caposino -S

Paula Caposino, Ph.D. Division 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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# Indications for Use

510(k) Number (if known) K241176

Device Name Alere NT-proBNP for Alinity i

The Alere NT-proBNP for Alinity i assay is a chemiluminescent microparticle immunoassay (CMIA) used for the in vitro quantitative determination of N-terminal pro B-type natriuretic peptide (NT-proBNP) in human serum and plasma on the Alinity i system.

In the emergency departments of NT-proBNP are used as an aid in the diagnosis of heart failure (HF) in patients with clinical suspicion of new onset or worsening HF.

| Type of Use (Select one or both, as applicable) | |
|------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------|
| <span> <span style="font-size: large;">☑</span> Prescription Use (Part 21 CFR 801 Subpart D) </span> | <span> ☐ Over-The-Counter Use (21 CFR 801 Subpart C) </span> |

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Indications for Use (Describe)

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#### Administrative Documentation - 510(k) Summary

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

#### I. 510(k) Number

K241176

# II. Applicant Name

Axis-Shield Diagnostics Limited Luna Place, The Technology Park, Dundee DD2 1XA United Kingdom

Primary contact:

Claire Dora, Associate Director, Regulatory Affairs Axis-Shield Diagnostics Ltd Phone: [phone] Email: [e-mail]

Date Summary Prepared: January 16, 2025

#### III. Device Name

Trade Name: Alere NT-proBNP for Alinity i

Alere NT-proBNP for Alinity i Device Classification: Class II Classification Name: BNP Test System Governing Regulation: 21 CFR Part 862.1117 Product Code: NBC

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#### IV. Predicate Device

Roche Elecsys proBNP II (K092649)

# V. Description of Device

# A. Reagents

The various configurations of the Alere NT-proBNP for Alinity i Reagent Kit are described below.

| | List Number | |
|------------------------------|-------------|---------|
| | 04S7921 | 04S7931 |
| Tests per cartridge | 100 | 500 |
| Number of cartridges per kit | 2 | 2 |
| Tests per kit | 200 | 1000 |
| Microparticle volume | 6.7 mL | 27.0 mL |
| Conjugate volume | 6.1 mL | 26.5 mL |

Microparticles: Anti-NT-proBNP (sheep, monoclonal) coated microparticles in Bis-TRIS buffer with protein (bovine) stabilizer, non-specific binding blocking agents, and surfactant. Minimum concentration: 0.05% solids. Preservative: sodium azide.

Conjugate: Anti-NT-proBNP (mouse, monoclonal) acridinium-labeled conjugate in MES buffer with protein (bovine) stabilizer and surfactant. Minimum concentration: 0.12 ug/mL. Preservatives: antimicrobial agents.

# B. Biological Principles of the Procedure

The Alere NT-proBNP for Alinity i assay is an automated, two-step immunoassay for the in vitro quantitative determination of NT-proBNP in human serum and plasma using chemiluminescent microparticle immunoassay (CMIA) technology.

Sample and anti-NT-proBNP coated paramagnetic microparticles are combined and incubated. The NT-proBNP present in the sample binds to the anti-NT-proBNP coated microparticles. The mixture is washed. Anti-NT-proBNP acridinium-labeled conjugate is added to create a reaction mixture and incubated. Following a wash cycle,

Pre-Trigger and Trigger Solutions are added.

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The resulting chemiluminescent reaction is measured as a relative light unit (RLU). There is a direct relationship between the amount of NT-proBNP in the sample and the RLU detected by the system optics.

#### Assay Standardization

There is currently no international recognized reference method or reference material for standardization. The Alere NT-proBNP for Alinity i Calibrators are traceable to internal reference standards at every concentration level that were value assigned on and made traceable to the predicate device by transference, as described by Clinical and Laboratory Standards Institute (CLSI) Document EP32-R.*

# C. Interpretation of Results

#### Emergency Department

For patients presenting to the emergency department (ED) with clinical suspicion of heart failure (HF), the results should be interpreted as indicated in the table below.

| Age Group<br>(Years) | NT-proBNP | | |
|----------------------|---------------------|-------------------|--------------------------------------------------|
| | (pg/mL) | (pmol/L) | Interpretation |
| All | < 300.0 | < 35.4 | Negative: HF unlikely |
| 18 to < 50 | ≥ 300.0 to < 450.0 | ≥ 35.4 to < 53.1 | Grayzone: Indeterminate |
| 50 to 75 | ≥ 300.0 to < 900.0 | ≥ 35.4 to < 106.2 | Consider other causes of<br>NT-proBNP elevation. |
| > 75 | ≥ 300.0 to < 1800.0 | ≥ 35.4 to < 212.4 | |
| 18 to < 50 | ≥ 450.0 | ≥ 53.1 | Positive: HF likely |
| 50 to 75 | ≥ 900.0 | ≥ 106.2 | |
| > 75 | ≥ 1800.0 | ≥ 212.4 | |

^&quot; Clinical and Laboratory Standards Institute (CLSI). Metrological Traceability and Its Implementation; A Report. CLSI document EP32-R. Wayne, PA: CLSI; 2006.

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# VI. Intended Use of the Device

The Alere NT-proBNP for Alinity i assay is a chemiluminescent microparticle immunoassay (CMIA) used for the in vitro quantitative determination of N-terminal pro B-type natriuretic peptide (NT-proBNP) in human serum and plasma on the Alinity i system.

In the emergency department, measurements of NT-proBNP are used as an aid in the diagnosis of heart failure (HF) in patients with clinical suspicion of new onset or worsening HF.

# VII. Comparison with the Predicate Device

The similarities and differences between the subject device and the predicate device are presented in the table below.

| | Subject Device:<br>Alere NT-proBNP for Alinity i | Predicate Device:<br>Roche Elecsys proBNP II (K092649) |
|----------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | General Device Characteristic Similarities | |
| Intended<br>Use/Indications<br>for Use | Immunoassay used for the in vitro<br>quantitative determination of N-terminal<br>pro B-type natriuretic peptide<br>(NT-proBNP) in human serum and<br>plasma.<br>In the emergency department,<br>measurements of NT-proBNP are used<br>as an aid in the diagnosis of heart failure<br>(HF) in patients with clinical suspicion<br>of new onset or worsening HF. | Immunoassay for the in vitro quantitative<br>determination of N-terminal pro-Brain<br>natriuretic peptide in human serum and<br>plasma.<br>This assay is used as an aid in the diagnosis of<br>individuals suspected of having congestive<br>heart failure.<br>The test is further indicated for the risk<br>stratification of patients with acute coronary<br>syndrome and congestive heart failure.<br>The test may also serve as an aid in the<br>assessment of increased risk of cardiovascular<br>events and mortality in patients at risk for<br>heart failure who have stable coronary artery<br>disease. |
| Specimen Type | Human serum and plasma | Same |
| Test Principle | Sandwich immunoassay | Same |
| Measurement<br>Type | Quantitative | Same |
| Analyte | NT-proBNP | Same |

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| | Subject Device:<br>Alere NT-proBNP for Alinity i | Predicate Device:<br>Roche Elecsys proBNP II (K092649) |
|----------------------------|------------------------------------------------------------------------------------------------------------------------------|---------------------------------------------------------------|
| Detection<br>Technology | Chemiluminescence | Same |
| General Device Differences | | |
| Principle of<br>Operation | CMIA | Electro chemiluminescent microparticle<br>immunoassay (ECLIA) |
| Intended Use<br>Setting | Emergency department | Not specified |
| Measuring<br>Interval | Analytical Measuring Interval (AMI):<br>15.8 to 35,000.0 pg/mL<br>(1.9 to 4130.0 pmoL/L) | 5 to 35,000 pg/mL |
| Hook Effect | No hook effect up to 372,620 pg/mL | No hook effect up to 300,000 pg/mL |
| Results<br>Interpretation | Negative: HF unlikely for all patients:<br>< 300.0 pg/mL | 125 pg/mL for < 75 years<br>450 pg/mL for ≥ 75 years |
| | Gray Zone: Indeterminate; consider other<br>causes of NT-proBNP elevation: | |
| | Age (Years) NT-proBNP (pg/mL)<br>18 to < 50 ≥ 300.0 to < 450.0<br>50 to 75 ≥ 300.0 to < 900.0<br>> 75 ≥ 300.0 to < 1800.0 | |
| | Positive: HF likely: | |
| | Age (Years) NT-proBNP (pg/mL)<br>18 to < 50 ≥ 450.0<br>50 to 75 ≥ 900.0<br>> 75 ≥ 1800.0 | |

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#### VIII. Summary of Nonclinical Performance

#### A. Reportable Interval

Based on representative data for the limit of quantitation (LoQ), the ranges over which results can be reported are provided below.

| | pg/mL | pmol/L |
|-----------------------------------------|----------------------|-------------------|
| Analytical Measuring<br>Interval (AMI)a | 15.8 - 35,000.0 | 1.9 - 4130.0 |
| Extended Measuring<br>Interval (EMI)b | 35,000.0 - 350,000.0 | 4130.0 - 41,300.0 |
| Reportable Intervalc | 15.8 - 350,000.0 | 1.9 - 41,300.0 |

ª AMI: The AMI extends from the LoQ to the upper limit of quantitation (ULoQ). This is determined by the range of values in pg/mL (pmol/L) that demonstrated acceptable performance for linearity, imprecision, and bias.

b EMI: The EMI extends from the ULoQ to the ULoQ × dilution factor. The value reflects a 1:10 dilution factor.

^ The reportable interval extends from the LoQ to the upper limit of the EMI.

# B. Within-Laboratory Precision (20-Day)

A study was performed based on guidance from CLSI EP05-A3. * Testing was conducted using 3 lots of the Alere NT-proBNP for Alinity i reagents, 1 lot of the Alere NT-proBNP for Alinity i Calibrators, 1 lot of the Alere NT-proBNP for Alinity i Controls, and 1 instrument. Three controls and 8 human plasma panels (7 native panels and 1 panel supplemented with NT-proBNP analyte) were tested in 2 replicates at 2 separate times per day on 20 days.

^* Clinical and Laboratory Standards Institute (CLS). Evaluation of Quantitative Measurement Procedures; Approved Guideline—Third Edition. CLSI Document EP05-A3. Wayne, PA: CLSI; 2014.

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| | | Mean | Repeatability | | Within-Laboratorya | | Between-Lot | | Overall Within-<br>Laboratoryb | |
|---------------------------|-----|----------|---------------|-----|--------------------|-----|-------------|-----|--------------------------------|------|
| Sample | n | (pg/mL) | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Low Control | 240 | 143.0 | 6.40 | 4.5 | 7.63 | 5.3 | 4.42 | 3.1 | 8.82 | 6.2 |
| Medium Control | 240 | 503.4 | 16.26 | 3.2 | 18.21 | 3.6 | 9.29 | 1.8 | 20.44 | 4.1 |
| High Control | 240 | 5144.2 | 177.02 | 3.4 | 202.18 | 3.9 | 21.47 | 0.4 | 203.32 | 4.0 |
| Panel A | 240 | 18.8 | 1.39 | 7.4 | 1.72 | 9.2 | 0.75 | 4.0 | 1.88 | 10.0 |
| Panel B | 240 | 55.6 | 2.64 | 4.7 | 3.02 | 5.4 | 1.60 | 2.9 | 3.41 | 6.1 |
| Panel C | 240 | 130.4 | 4.28 | 3.3 | 5.44 | 4.2 | 4.87 | 3.7 | 7.30 | 5.6 |
| Panel D | 240 | 449.1 | 13.10 | 2.9 | 18.68 | 4.2 | 14.02 | 3.1 | 23.36 | 5.2 |
| Panel E | 240 | 1011.4 | 28.31 | 2.8 | 34.49 | 3.4 | 21.92 | 2.2 | 40.87 | 4.0 |
| Panel F | 240 | 1938.0 | 54.02 | 2.8 | 65.28 | 3.4 | 22.56 | 1.2 | 69.07 | 3.6 |
| Panel G | 240 | 14,939.8 | 417.56 | 2.8 | 518.88 | 3.5 | 296.61 | 2.0 | 597.67 | 4.0 |
| Panel H<br>(Supplemented) | 240 | 31,711.0 | 989.02 | 3.1 | 1185.11 | 3.7 | 2118.19 | 6.7 | 2427.19 | 7.7 |

ª Includes repeatability (within-run), between-run, and between-day variability.

b Includes repeatability (within-run), between-run, between-day, and between-lot variability.

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#### C. Linearity

A study was performed based on guidance from CLSI EP06, 2nd ed. This assay is linear across the AMI of 15.8 to 35,000.0 pg/mL (1.9 to 4130.0 pmol/L).

#### D. Lower Limits of Measurement

A study was performed based on guidance from CLSI EP17-A2. T Testing was conducted using 3 lots of the Alere NT-proBNP for Alinity i reagents on each of 2 instruments over a minimum of 3 days. The maximum observed limit of blank (LoB), limit of detection (LoD), and limit of quantitation (LoQ) values are summarized below.

| | pg/mL | pmol/L |
|------|-------|--------|
| LoBa | 0.1 | 0.0 |
| LoDb | 3.6 | 0.4 |
| LoQc | 15.8 | 1.9 |

a The LoB represents the 95th percentile from n ≥ 60 replicates of zero-analyte samples.

b The LoD represents the lowest concentration at which the analyte can be detected with 95% probability based on n ≥ 60 replicates of low-analyte level samples.

6 The LoQ is defined as the lowest concentration at which a maximum allowable precision of 20 %CV was met and was determined from n ≥ 60 replicates of low-analyte level samples.

# E. Analytical Specificity

#### 1. Potentially Interfering Endogenous Substances

A study was performed based on guidance from CLSI EP07, 3rd ed. + Each substance was tested at 2 levels of the analyte (approximately 125 pg/mL and 2000 pg/mL).

^* Clinical and Laboratory Standards Institute (CLSI). Evaluation of the Linearity of Quantitative Measurement Procedures. 2nd ed. CLSI Guideline EP06. Wayne, PA: CLSI; 2020.

T Clinical and Laboratory Standards Institute (CLSI). Evaluation of Detection Capability for Clinical Measurements; Approved Guideline-Second Edition. CLSI Document EP17-A2. Wayne, PA: CLSI; 2012.

⁺ Clinical and Laboratory Standards Institute (CLSI). Interference Testing in Clinical Chemistry. 3rd ed. CLSI Guideline EP07. Wayne, PA: CLSI; 2018.

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No significant interference (interference within ± 10.0%) was observed at the following concentrations.

| No Significant Interference (Interference within ± 10.0%) | |
|-----------------------------------------------------------|-------------------|
| Potentially Interfering Substance | Interferent Level |
| Bilirubin (conjugated) | 60 mg/dL |
| Bilirubin (unconjugated) | 60 mg/dL |
| Biotin | 4250 ng/mL |
| Cholesterol | 700 mg/dL |
| Human Anti-mouse Antibodies (HAMA) | 1500 ng/mL |
| Hemoglobin | 1 g/dL |
| IgG | 6 g/dL |
| Intralipid | 3000 mg/dL |
| Rheumatoid Factor (RF) | 600 IU/mL |
| Total Protein | 12.6 g/dL |

#### Interference beyond ± 10.0% (based on 95% confidence interval [CI]) was

observed at the concentrations shown below for the following substances.

| Interference beyond ± 10.0% (based on 95% CI) | | | |
|-----------------------------------------------|-------------------|---------------|----------------------------|
| Potentially Interfering<br>Substance | Interferent Level | Analyte Level | % Interference<br>(95% CI) |
| RF | 1520 IU/mL | 125 pg/mL | -8.9%<br>(-10.4%, -7.5%) |
| | 1520 IU/mL | 2000 pg/mL | -11.4%<br>(-12.4%, -10.4%) |
| Total Protein | 15.2 g/dL | 125 pg/mL | -12.7%<br>(-14.7%, -10.7%) |
| | 15.5 g/dL | 2000 pg/mL | -9.9%<br>(-11.4%, -8.5%) |

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# 2. Potentially Interfering Drugs

A study was performed based on guidance from CLSI EP07, 3rd ed. * Each substance was tested at 2 levels of the analyte (approximately 125 pg/mL and 2000 pg/mL).

No significant interference (interference within ± 10.0%) was observed at the following concentrations.

| No Significant Interference (Interference within ± 10.0%) | | | |
|-----------------------------------------------------------|-------------------|---------------------------------|-------------------|
| Potentially Interfering<br>Drug | Interferent Level | Potentially Interfering<br>Drug | Interferent Level |
| Acetaminophen | 15.6 mg/dL | Lidocaine | 8.0 mg/dL |
| Acetylsalicylic Acid | 60 mg/dL | Lisinopril | 1.6 mg/dL |
| Allopurinol | 6.0 mg/dL | Lithium | 4.20 mg/dL |
| Amikacin | 15 mg/dL | Losartan potassium | 5.99 mg/dL |
| Amiodarone | 4.2 mg/dL | Lovastatin | 2.0 mg/dL |
| Amlodipine Besylate | 0.4 mg/dL | L-Thyroxine | 0.06 mg/dL |
| Ampicillin | 7.5 mg/dL | Methyldopa | 2.5 mg/dL |
| Ascorbic acid | 5.25 mg/dL | Methylprednisolone | 0.75 mg/dL |
| Atenolol | 1.0 mg/dL | Metoprolol | 1.5 mg/dL |
| Atorvastatin | 32 mg/dL | Milrinone | 0.183 mg/dL |
| Caffeine | 10.8 mg/dL | Naproxen | 49.97 mg/dL |
| Captopril | 5.0 mg/dL | Nicotine | 0.1 mg/dL |
| Carbamazepine | 4.5 mg/dL | Nicotinic acid | 4.0 mg/dL |
| Carvedilol | 3.75 mg/dL | Nifedipine | 6.0 mg/dL |
| Chloramphenicol | 7.8 mg/dL | Nitrofurantoin | 4.0 mg/dL |
| Chlordiazepoxide | 1.0 mg/dL | Nitroglycerin | 0.016 mg/dL |
| Chlorpromazine | 0.33 mg/dL | Oxazepam | 1.2 mg/dL |
| Cimetidine | 3.0 mg/dL | Oxytetracycline | 10 mg/dL |
| No Significant Interference (Interference within ± 10.0%) | | | |
| Potentially Interfering<br>Drug | Interferent Level | Potentially Interfering<br>Drug | Interferent Level |
| Cinnarizine | 3.1 mg/dL | Penicillin G | 25 U/mL |
| Clopidogrel bisulfate | 7.5 mg/dL | Pentobarbital | 12.6 mg/dL |
| Creatinine | 30 mg/dL | Phenobarbitol | 69 mg/dL |
| Cyclosporine A | 0.4 mg/dL | Phenprocoumon<br>(Marcumar) | 1.53 mg/dL |
| Dextran 40 | 6000 mg/dL | Phenytoin | 6.0 mg/dL |
| Diazepam | 3.0 mg/dL | Primidone | 5.7 mg/dL |
| Diclofenac | 6.0 mg/dL | Probenecid | 60 mg/dL |
| Digoxin | 0.025 mg/dL | Procainamide | 4.8 mg/dL |
| Diltiazem | 12 mg/dL | Propafenone | 30 mg/dL |
| Dipyridamole | 8.0 mg/dL | Propanolol | 0.2 mg/dL |
| Disopyramide | 4.0 mg/dL | Propoxyphene | 0.321 mg/dL |
| Dobutamine | 10 mg/dL | Quinidine | 2.0 mg/dL |
| Dopamine | 65 mg/dL | Ramipril | 0.6 mg/dL |
| Enalapril Maleate | 1.6 mg/dL | Retaplase | 3.33 mg/dL |
| Epinephrine | 0.05 mg/dL | Simvastatin | 3.2 mg/dL |
| Erythromycin | 13.8 mg/dL | Spironolactone | 7.5 mg/dL |
| Ethanol | 600 mg/dL | Sulfamethoxazole | 112 mg/dL |
| Ethosuximide | 30 mg/dL | Theophylline | 6.0 mg/dL |
| Fenofibrate | 4.5 mg/dL | Tolbutamide | 150 mg/dL |
| Furosemide | 6.0 mg/dL | Torasemide | 1.5 mg/dL |
| Gentamicin | 12 mg/dL | Trandolapril | 4.0 mg/dL |
| Heparin | 8 U/mL | Trasylol/Aprotinin | 501.8 KIE/mL |
| Hydralazine | 2.0 mg/dL | Trimethoprim | 6.4 mg/dL |
| Hydrochlorothiazide | 2.0 mg/dL | Uric Acid | 23.5 mg/dL |
| Insulin | 0.16 mg/dL | Valproic Acid | 50 mg/dL |
| Ibuprofen | 50 mg/dL | Verapamil | 24 mg/dL |
| No Significant Interference (Interference within ± 10.0%) | | | |
| Potentially Interfering<br>Drug | Interferent Level | Potentially Interfering<br>Drug | Interferent Level |
| Indomethacin | 3.6 mg/dL | Warfarin | 7.5 mg/dL |
| Isosorbide dinitrate | 6.0 mg/dL | | |

^* Clinical and Laboratory Standards Institute (CLSI). Interference Testing in Clinical Chemistry. 3rd ed. CLSI Guideline EP07. Wayne, PA: CLSI; 2018.

{14}------------------------------------------------

{15}------------------------------------------------

#### 3. Cross-Reactants

A study was performed based on guidance from CLSI EP07, 3rd ed. * Samples with NT-proBNP target concentrations of 125 pg/mL and 2000 pg/mL containing the cross-reactants at the concentration listed below were tested with the Alere NT-proBNP for Alinity i assay on the Alinity i system. For each cross-reactant, the % recovery was calculated as: (NT-proBNP concentration with cross-reactant) / (NT-proBNP concentration without cross-reactant) × 100%. The observed % recovery of NT-proBNP was within 100% ± 10% for all cross-reactants evaluated at each analyte level.

| Potential Cross-Reactant | Cross-Reactant Concentration |
|---------------------------------|------------------------------|
| Adrenomedullin | 1000 pg/mL |
| Aldosterone | 600 pg/mL |
| Angiotensin I | 600 pg/mL |
| Angiotensin II | 600 pg/mL |
| Angiotensin III | 1000 pg/mL |
| ANP 28 | 3100 ng/mL |
| Arg-Vasopressin | 1000 pg/mL |
| BNP 32 | 3500 ng/mL |
| CNP 22 | 2200 ng/mL |
| Endothelin | 20 pg/mL |
| NT-proANP 1-30 (preproANP26-55) | 3500 ng/mL |

^* Clinical and Laboratory Standards Institute (CLSI). Interference Testing in Clinical Chemistry. 3rd ed. CLSI Guideline EP07. Wayne, PA: CLSI; 2018.

{16}------------------------------------------------

| Potential Cross-Reactant | Cross-Reactant Concentration |
|------------------------------------|------------------------------|
| NT-proANP 31-67 (preproANP56-92) | 1000 pg/mL |
| NT-proANP 79-98 (preproANP104-123) | 1000 pg/mL |
| Renin | 50,000 pg/mL |
| Urodilatin | 3500 ng/mL |

#### F. High Dose Hook

High dose hook effect was not observed on samples up to 372,620 pg/mL.

#### G. Expected Values (Reference Interval)

A reference interval study was performed based on guidance from CLSI EP28-A3c. Testing was performed with a US-based general population from apparently healthy individuals (≥ 18 years old). The specimens were tested with the Alere NT-proBNP for Alinity i assay on the Alinity i system. Based on the results, the 95% reference interval of an apparently healthy population for each sex and age range was determined to be as follows:

| | | | | Reference Interval |
|----------------------|--------|-----|---------|------------------------------|
| | | | Median | (2.5th to 97.5th Percentile) |
| Age Range | Sex | n | (pg/mL) | (pg/mL) |
| 18 to < 50 years old | Female | 181 | 30.7 | < 15.8 to 104.8 |
| | Male | 137 | < 15.8 | < 15.8 to 180.3 |
| 50 to 75 years old | Female | 134 | 51.7 | <15.8 to 334.1 |
| | Male | 146 | 39.9 | < 15.8 to 451.6 |
| > 75 years old | Female | 127 | 81.4 | < 15.8 to 956.1 |
| | Male | 136 | 65.1 | < 15.8 to 683.0 |

Clinical and Laboratory Standards Institute (CLSI). Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory; Approved Guideline—Third Edition. CLSI document EP28-A3c. Wayne, PA: CLSI; 2008.

{17}------------------------------------------------

# IX. Summary of Clinical Performance

#### A. Reproducibility

A study was performed based on guidance from CLSI EP05-A3. * Testing was conducted at each of 3 testing sites using 3 lots of the Alere NT-proBNP for Alinity i reagents, 2 lots of the Alere NT-proBNP for Alinity i Calibrators, 2 lots of the Alere NT-proBNP for Alinity i Controls, and 1 instrument. Three controls and 8 human plasma panels (6 native panels and 2 panels supplemented with NT-proBNP analyte) were tested in a minimum of 3 replicates at 2 separate times per day on 5 different days.

^* Clinical and Laboratory Standards Institute (CLSI). Evaluation of Quantitative Measurement Procedures; Approved Guideline-Third Edition. CLSI Document EP05-A3. Wayne, PA: CLSI; 2014.

{18}------------------------------------------------

| Sample | n | Mean<br>(pg/mL) | Repeatability<br>SD | Repeatability<br>%CV | Within-<br>Laboratorya<br>SD | Within-<br>Laboratorya<br>%CV | Between-Site<br>SD | Between-Site<br>%CV | Between-Lot<br>SD | Between-Lot<br>%CV | Overall<br>Reproducibilityb<br>SD | Overall<br>Reproducibilityb<br>%CV |
|---------------------------|-----|-----------------|---------------------|----------------------|------------------------------|-------------------------------|--------------------|---------------------|-------------------|--------------------|-----------------------------------|------------------------------------|
| Low Control | 360 | 136.6 | 4.75 | 3.5 | 5.75 | 4.2 | 2.63 | 1.9 | 1.45 | 1.1 | 6.49 | 4.7 |
| Medium Control | 360 | 477.1 | 12.24 | 2.6 | 15.35 | 3.2 | 14.20 | 3.0 | 8.88 | 1.9 | 22.72 | 4.8 |
| High Control | 359 | 4932.2 | 201.01 | 4.1 | 311.08 | 6.3 | 105.17 | 2.1 | 0.00 | 0.0 | 328.38 | 6.7 |
| Panel 1 | 359 | 14.5 | 2.10c | 14.5 | 2.74c | 18.9 | 0.00 | 0.0 | 0.00 | 0.0 | 2.74c | 18.9 |
| Panel 2 | 360 | 51.9 | 2.14 | 4.1 | 3.07 | 5.9 | 0.49 | 0.9 | 0.00 | 0.0 | 3.11 | 6.0 |
| Panel 3 | 360 | 127.1 | 4.67 | 3.7 | 5.95 | 4.7 | 1.36 | 1.1 | 0.78 | 0.6 | 6.15 | 4.8 |
| Panel 4 | 360 | 436.9 | 10.46 | 2.4 | 18.67 | 4.3 | 3.18 | 0.7 | 0.00 | 0.0 | 18.94 | 4.3 |
| Panel 5 | 360 | 996.5 | 21.86 | 2.2 | 43.45 | 4.4 | 17.12 | 1.7 | 0.00 | 0.0 | 46.70 | 4.7 |
| Panel 6 | 360 | 1891.1 | 33.18 | 1.8 | 79.86 | 4.2 | 35.96 | 1.9 | 0.00 | 0.0 | 87.58 | 4.6 |
| Panel 7<br>(Supplemented) | 359 | 15,664.6 | 319.91 | 2.0 | 974.61 | 6.2 | 281.81 | 1.8 | 217.28 | 1.4 | 1037.54 | 6.6 |
| Panel 8<br>(Supplemented) | 360 | 25,630.5 | 601.42 | 2.3 | 1654.86 | 6.5 | 220.39 | 0.9 | 807.19 | 3.1 | 1854.37 | 7.2 |

Includes repeatability (within-run), between-run, and between-day variability. a

b Includes repeatability (within-run), between-run, between-site, and between-lot variability.

င An outlying run was observed. Based on guidance from CLSI EPO5-A3, a replacement run was performed and the results are shown in the preceding table. Without the replacement run, the repeatability SD was 9.5 pg/mL (1.12 pmol2), within-laboratory precision SD was 9.64 pg/mL (1.14 pmoVL), and the overall reproducibility SD was 9.64 pg/mL (1.14 pmol/L).

{19}------------------------------------------------

#### B. Clinical Performance

A multi-center prospective study including 17 collection sites across the US was conducted to establish the performance characteristics of the Alere NT-proBNP for Alinity i assay. Subjects 18 years and older presenting to the ED with signs and symptoms consistent with a clinical suspicion of new onset or acute exacerbation of HF were included in the study. Subjects with renal insufficiency requiring dialysis or known estimated glomerular filtration rate (eGFR) < 15.0 mL/min/1.73 m² and subjects with dyspnea after chest trauma were excluded from the study. An adjudicated diagnosis was determined by a panel of board-certified cardiologists.

The results were determined from 2127 ED subjects. Of the 2127 ED subjects, 1030 (48.4%) were female and 1097 (51.6%) were male, ranging in age from 19 to 97 years. Individuals in the population were White (53.1%), Black or African American (39.5%), Asian (1.7%), American Indian or Alaska Native (0.8%), Native Hawaiian or Other Pacific Islander (0.3%), or represented by other races (0.9%), with the remaining 3.6% of Unknown / Not Reported race. A total of 1934 (90.9%) ED subjects were not Hispanic or Latino, and 168 (7.9%) ED subjects were Hispanic or Latino, with the remaining 25 (1.2%) ED subjects of Unknown / Not Reported ethnicity.

The descriptive statistics for the Alere NT-proBNP for Alinity i test results (pg/mL) were determined within and across sex by age group and are summarized in the following table.

| Adjudicated<br>Diagnosis | HF | | | | Non-HF | | | |
|--------------------------|---------|----------|---------|---------|---------|----------|---------|---------|
| Age Group<br>(Years) | < 50 | 50 to 75 | > 75 | All | < 50 | 50 to 75 | > 75 | All |
| All Subjects | | | | | | | | |
| N | 160 | 553 | 167 | 880 | 300 | 799 | 148 | 1247 |
| Mean | 4354.5 | 6092.9 | 6891.3 | 5928.3 | 437.9 | 760.4 | 1738.8 | 798.9 |
| SD | 5454.61 | 8144.69 | 6860.32 | 7521.45 | 1686.57 | 1833.48 | 3494.96 | 2100.93 |
| Median | 2664.9 | 3297.2 | 4787.6 | 3420.7 | 70.1 | 198.9 | 565.9 | 181.8 |

{20}------------------------------------------------

| Adjudicated<br>Diagnosis | HF | | | | Non-HF | | | |
|--------------------------|----------|----------|----------|----------|----------|----------|----------|----------|
| Age Group<br>(Years) | < 50 | 50 to 75 | > 75 | All | < 50 | 50 to 75 | > 75 | All |
| Minimum | 22.8 | 19.8 | 158.9 | 19.8 | 0.0 | 0.0 | 4.2 | 0.0 |
| Maximum | 38,880.9 | 60,716.9 | 38,938.4 | 60,716.9 | 24,737.1 | 17,988.9 | 31,154.0 | 31,154.0 |
| Female Subjects | | | | | | | | |
| N | 55 | 236 | 85 | 376 | 165 | 407 | 82 | 654 |
| Mean | 4034.7 | 5370.1 | 6814.2 | 5501.2 | 273.5 | 614.8 | 1320.8 | 617.2 |
| SD | 6302.82 | 7732.10 | 7042.32 | 7417.01 | 732.64 | 1543.13 | 2403.63 | 1556.92 |
| Median | 1851.5 | 2541.7 | 4787.6 | 2786.4 | 73.1 | 174.9 | 499.7 | 161.2 |
| Minimum | 22.8 | 24.9 | 158.9 | 22.8 | 0.0 | 0.9 | 25.7 | 0.0 |
| Maximum | 38,880.9 | 47,385.1 | 38,938.4 | 47,385.1 | 5633.9 | 14,849.5 | 18,827.7 | 18,827.7 |
| Male Subjects | | | | | | | | |
| N | 105 | 317 | 82 | 504 | 135 | 392 | 66 | 593 |
| Mean | 4522.0 | 6631.0 | 6971.2 | 6247.0 | 638.8 | 911.6 | 2258.0 | 999.4 |
| SD | 4977.78 | 8410.52 | 6708.84 | 7590.10 | 2369.85 | 2084.06 | 4464.35 | 2557.26 |
| Median | 2780.1 | 4066.5 | 4773.9 | 3904.3 | 64.1 | 221.0 | 701.8 | 203.2 |
| Minimum | 58.3 | 19.8 | 186.9 | 19.8 | 0.0 | 0.0 | 4.2 | 0.0 |
| Maximum | 26,986.3 | 60,716.9 | 32,286.4 | 60,716.9 | 24,737.1 | 17,988.9 | 31,154.0 | 31,154.0 |

The pretest probability of adjudicated HF (prevalence of adjudicated HF in the study), posttest probabilities, and likelihood ratios of the Alere NT-proBNP for Alinity i result vs adjudicated diagnosis (along with the 95% CIs) were determined based on guidance from CLSI EP12, 3rd ed., * for all subjects and by sex using the age-dependent positive cutoffs (450 pg/mL for subjects 18 to < 50 years of age, 900 pg/mL for subjects 50 to 75 years of age, and 1800 pg/mL for subjects > 75 years of age) and age-independent negative cutoff (300 pg/mL).

^* Clinical and Laboratory Standards Institute (CLSI). Evaluation of Qualitative, Binary Output Examination Performance. 3rd ed. CLSI Guideline EP12. Wayne, PA: CLSI; 2023.

{21}------------------------------------------------

| | | | All Subjects | | | | | |
|--------------------------------------------------------------------------------------------------------|-----|--------|--------------------------|--|--|-------------------------------------------------------|--------------------------------------------------------------|---------------------------------------|
| NT-proBNP<br>Result | | | Adjudicated<br>Diagnosis | | | Posttest<br>Probability of<br>HF % (n/N)<br>(95% CI)a | Posttest<br>Probability of<br>Non-HF %<br>(n/N)<br>(95% CI)a | Likelihood<br>Ratio (HF)<br>(95% CI)b |
| | HF | Non-HF | Total | | | | | |
| Age Group: 18 to < 50 Years<br>Pretest Probability of HF (Prevalence of HF in Study): 34.78% (160/460) | | | | | | | | |
| Positive | 140 | 48 | 188 | | | 74.5<br>(140/188)<br>(67.8, 80.2) | - | 5.47<br>(4.19, 7.13) |
| Grayzone | 7 | 13 | 20 | | | 35.0<br>(7/20)<br>(18.1, 56.7) | 65.0<br>(13/20)<br>(43.3, 81.9) | 1.01<br>(0.41, 2.48) |
| Negative | 13 | 239 | 252 | | | - | 94.8<br>(239/252)<br>(91.4, 97.0) | 0.10<br>(0.06, 0.17) |
| Total | 160 | 300 | 460 | | | | | |
| Age Group: 50 to 75 Years<br>Pretest Probability of HF (Prevalence of HF in Study): 40.90% (553/1352) | | | | | | | | |
| Positive | 437 | 148 | 585 | | | 74.7<br>(437/585)<br>(71.0, 78.1) | - | 4.27<br>(3.67, 4.96) |
| Grayzone | 80 | 147 | 227 | | | 35.2<br>(80/227)<br>(29.3, 41.7) | 64.8<br>(147/227)<br>(58.3, 70.7) | 0.79<br>(0.61, 1.01) |
| Negative | 36 | 504 | 540 | | | - | 93.3<br>(504/540)<br>(90.9, 95.1) | 0.10<br>(0.07, 0.14) |
| Total | 553 | 799 | 1352 | | | | | |

The results for all subjects are presented in the following table.

Age Group: > 75 Years

Pretest Probability of HF (Prevalence of HF in Study): 53.02% (167/315)

{22}------------------------------------------------

| | | | | All Subjects | | |
|--------------------------------------------------------------------------|-----------------------|--------|-------|-------------------------------------------------------|--------------------------------------------------------------|---------------------------------------|
| | Adjudicated Diagnosis | | | Posttest<br>Probability of<br>HF % (n/N)<br>(95% CI)a | Posttest<br>Probability of<br>Non-HF %<br>(n/N)<br>(95% CI)a | Likelihood<br>Ratio (HF)<br>(95% CI)b |
| NT-proBNP<br>Result | HF | Non-HF | Total | | | |
| Positive | 131 | 38 | 169 | 77.5<br>(131/169)<br>(70.6, 83.2) | - | 3.06<br>(2.30, 4.06) |
| Grayzone | 34 | 59 | 93 | 36.6<br>(34/93)<br>(27.5, 46.7) | 63.4<br>(59/93)<br>(53.3, 72.5) | 0.51<br>(0.36, 0.73) |
| Negative | 2 | 51 | 53 | - | 96.2<br>(51/53)<br>(87.2, 99.0) | 0.03<br>(0.01, 0.14) |
| Total | 167 | 148 | 315 | | | |
| All Subjectsc | | | | | | |
| Pretest Probability of HF (Prevalence of HF in Study): 41.37% (880/2127) | | | | | | |
| Positive | 708 | 234 | 942 | 75.2<br>(708/942)<br>(72.3, 77.8) | - | 4.29<br>(3.80, 4.83) |
| Grayzone | 121 | 219 | 340 | 35.6<br>(121/340)<br>(30.7, 40.8) | 64.4<br>(219/340)<br>(59.2, 69.3) | 0.78<br>(0.64, 0.96) |
| Negative | 51 | 794 | 845 | - | 94.0<br>(794/845)<br>(92.2, 95.4) | 0.09<br>(0.07, 0.12) |
| Total | 880 | 1247 | 2127 | | | |

The results for female subjects are presented in the following table.

{23}------------------------------------------------

| | | | Female Subjects | | | |
|--------------------------------------------------------------------------|-------------------------------------------------------------------------|--------|-----------------|-------------------------------------------------------|--------------------------------------------------------------|---------------------------------------|
| NT-proBNP<br>Result | Adjudicated<br>Diagnosis | | Total | Posttest<br>Probability of<br>HF % (n/N)<br>(95% CI)a | Posttest<br>Probability of<br>Non-HF %<br>(n/N)<br>(95% CI)a | Likelihood<br>Ratio (HF)<br>(95% CI)b |
| | HF | Non-HF | | | | |
| Age Group: 18 to < 50 Years | | | | | | |
| | Pretest Probability of HF (Prevalence of HF in Study): 25.00% (55/220) | | | | | |
| Positive | 45 | 20 | 65 | 69.2<br>(45/65)<br>(57.2, 79.1) | - | 6.75<br>(4.39, 10.37) |
| Grayzone | 3 | 7 | 10 | 30.0<br>(3/10)<br>(10.8, 60.3) | 70.0<br>(7/10)<br>(39.7, 89.2) | 1.29<br>(0.34, 4.80) |
| Negative | 7 | 138 | 145 | - | 95.2<br>(138/145)<br>(90.4, 97.6) | 0.15<br>(0.08, 0.31) |
| Total | 55 | 165 | 220 | | | |
| Age Group: 50 to 75 Years | | | | | | |
| | Pretest Probability of HF (Prevalence of HF in Study): 36.70% (236/643) | | | | | |
| Positive | 173 | 56 | 229 | 75.5<br>(173/229)<br>(69.6, 80.7) | - | 5.33<br>(4.13, 6.88) |
| Grayzone | 35 | 76 | 111 | 31.5<br>(35/111)<br>(23.6, 40.7) | 68.5<br>(76/111)<br>(59.3, 76.4) | 0.79<br>(0.55, 1.15) |
| Negative | 28 | 275 | 303 | - | 90.8<br>(275/303)<br>(87.0, 93.5) | 0.18<br>(0.12, 0.25) |
| Total | 236 | 407 | 643 | | | |
| Age Group: > 75 Years | | | | | | |
| | Pretest Probability of HF (Prevalence of HF in Study): 50.90% (85/167) | | | | | |
| Positive | 64 | 19 | 83 | 77.1<br>(64/83)<br>(67.0, 84.8) | - | 3.25<br>(2.15, 4.91) |
| Female Subjects | | | | | | |
| | Adjudicated<br>Diagnosis | | | | | |
| NT-proBNP<br>Result | HF | Non-HF | Total | Posttest<br>Probability of<br>HF % (n/N)<br>(95% CI)a | Posttest<br>Probability of<br>Non-HF %<br>(n/N)<br>(95% CI)a | Likelihood<br>Ratio (HF)<br>(95% CI)b |
| Grayzone | 20 | 34 | 54 | 37.0<br>(20/54)<br>(25.4, 50.4) | 63.0<br>(34/54)<br>(49.6, 74.6) | 0.57<br>(0.36, 0.90) |
| Negative | 1 | 29 | 30 | - | 96.7<br>(29/30)<br>(83.3, 99.4) | 0.03<br>(0.00, 0.24) |
| Total | 85 | 82 | 167 | | | |
| All Female Subjectsc | | | | | | |
| Pretest Probability of HF (Prevalence of HF in Study): 36.50% (376/1030) | | | | | | |
| Positive | 282 | 95 | 377 | 74.8<br>(282/377)<br>(70.2, 78.9) | - | 5.16<br>(4.25, 6.27) |
| Grayzone | 58 | 117 | 175 | 33.1<br>(58/175)<br>(26.6, 40.4) | 66.9<br>(117/175)<br>(59.6, 73.4) | 0.86<br>(0.65, 1.15) |
| Negative | 36 | 442 | 478 | - | 92.5<br>(442/478)<br>(89.7, 94.5) | 0.14<br>(0.10, 0.19) |
| Total | 376 | 654 | 1030 | | | |

Administrative Documentation – 510(k) Summary
Administrative Documentation – 510(k) Summary
Page 20 of 42

{24}------------------------------------------------

The results for male subjects are presented in the following table.

{25}------------------------------------------------

| | Male Subjects | | | | | | | | | |
|-----------------------------|--------------------------|------------|-------|-------------------------------------------------------------------------|----------------------------------------|-------------------------|--|--|--|--|
| NT-proBNP<br>Result | Adjudicated<br>Diagnosis | | | Posttest<br>Probability of | Posttest<br>Probability of<br>Non-HF % | Likelihood | | | | |
| | HF | Non-<br>HF | Total | HF % (n/N)<br>(95% CI)a | (n/N)<br>(95% CI)a | Ratio (HF)<br>(95% CI)b | | | | |
| Age Group: 18 to < 50 Years | | | | | | | | | | |
| | | | | Pretest Probability of HF (Prevalence of HF in Study): 43.75% (105/240) | | | | | | |
| Positive | 95 | 28 | 123 | 77.2<br>(95/123)<br>(69.1, 83.8) | - | 4.36<br>(3.12, 6.10) | | | | |
| Grayzone | 4 | 6 | 10 | 40.0<br>(4/10)<br>(16.8, 68.7) | 60.0<br>(6/10)<br>(31.3, 83.2) | 0.86<br>(0.25, 2.96) | | | | |
| Negative | 6 | 101 | 107 | - | 94.4<br>(101/107)<br>(88.3, 97.4) | 0.08<br>(0.03, 0.17) | | | | |
| Total | 105 | 135 | 240 | | | | | | | |
| Age Group: 50 to 75 Years | | | | | | | | | | |
| | | | | Pretest Probability of HF (Prevalence of HF in Study): 44.71% (317/709) | | | | | | |
| Positive | 264 | 92 | 356 | 74.2<br>(264/356)<br>(69.4, 78.4) | - | 3.55<br>(2.95, 4.27) | | | | |
| Grayzone | 45 | 71 | 116 | 38.8<br>(45/116)<br>(30.4, 47.9) | 61.2<br>(71/116)<br>(52.1, 69.6) | 0.78<br>(0.56, 1.10) | | | | |
| Negative | 8 | 229 | 237 | - | 96.6<br>(229/237)<br>(93.5, 98.3) | 0.04<br>(0.02, 0.09) | | | | |
| Total | 317 | 392 | 709 | | | | | | | |
| Age Group: > 75 Years | | | | | | | | | | |
| | | | | Pretest Probability of HF (Prevalence of HF in Study): 55.41% (82/148) | | | | | | |
| Positive | 67…