i-STAT TBI Plasma cartridge with the i-STAT Alinity System

K201778 · Abbott Laboratories · QAT · Jan 8, 2021 · Immunology

Device Facts

• Record ID: K201778
• Device Name: i-STAT TBI Plasma cartridge with the i-STAT Alinity System
• Applicant: Abbott Laboratories
• Product Code: QAT · Immunology
• Decision Date: Jan 8, 2021
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.5830
• Device Class: Class 2
• Attributes: AI/ML

Intended Use

The i-STAT TBI Plasma test is a panel of in vitro diagnostic immunoassays for the quantitative measurements of glial fibrillary acidic protein (GFAP) and ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) in plasma and a semiquantitative interpretation of test results derived from these measurements, using the i-STAT Alinity Instrument. The interpretation of test results is used, in conjunction with other clinical information, to aid in the evaluation of patients, 18 years of age or older, presenting with suspected mild traumatic brain injury (Glasgow Coma Scale score 13-15) within 12 hours of injury, to assist in determining the need for a CT (computed tomography) scan of the head. A 'Not Elevated' test interpretation is associated with the absence of acute traumatic intracranial lesions visualized on a head CT scan. The test is to be used with plasma prepared from EDTA anticoagulated specimens in clinical laboratory settings by a healthcare professional. The i-STAT TBI Plasma test is not intended to be used in point of care settings.

Device Story

The i-STAT TBI Plasma cartridge is a multiplex immunoassay for quantitative measurement of GFAP and UCH-L1 biomarkers in EDTA plasma. Used with the i-STAT Alinity instrument in clinical laboratory settings by healthcare professionals. The cartridge is inserted into the handheld i-STAT Alinity instrument, which acts as an electro-mechanical interface; the instrument executes the test cycle, acquires electrical sensor signals, and converts them into quantitative results via a microprocessor. The system provides a semi-quantitative interpretation of test results within approximately 15 minutes. Results are used alongside clinical information to aid in evaluating patients with suspected mild TBI to determine the necessity of a head CT scan. A 'Not Elevated' result correlates with the absence of acute traumatic intracranial lesions on CT, potentially reducing unnecessary head CT scans. The device offers workflow efficiencies compared to the predicate ELISA-based method.

Clinical Evidence

Clinical performance evaluated in two studies: 1) Retrospective analysis of 1901 frozen plasma samples from ALERT-TBI study (sensitivity 95.8%, specificity 40.4%, NPV 99.3%); 2) Prospective validation with 88 fresh plasma samples from TRACK-TBI Phase 2 study (sensitivity 100%, specificity 23.7%, NPV 100%).

Technological Characteristics

Multiplex immunoassay cartridge using electrochemical detection. Measures GFAP (30–10,000 pg/mL) and UCH-L1 (200–3,200 pg/mL). Designed for use with the i-STAT Alinity handheld instrument. Requires EDTA plasma. Calibration is pre-set during manufacture. No user calibration required.

Indications for Use

Indicated for patients 18+ years old with suspected mild traumatic brain injury (GCS 13-15) within 12 hours of injury to assist in determining the need for a head CT scan. Not for point-of-care use.

Regulatory Classification

Identification

A brain trauma assessment test is a device that consists of reagents used to detect and measure brain injury biomarkers in human specimens. The measurements aid in the evaluation of patients with suspected mild traumatic brain injury in conjunction with other clinical information to assist in determining the need for head imaging per current standard of care.

Special Controls

*Classification.* Class II (special controls). The special controls for this device are:(1) The 21 CFR 809.10(b) compliant labeling must include detailed descriptions of and results from performance testing conducted to evaluate precision, accuracy, linearity, analytical sensitivity, interference, and cross-reactivity. This information must include the following: (i) Performance testing of device precision must, at minimum, use one unmodified clinical specimen from the intended use population with concentration of the brain injury biomarker(s) near the medical decision point. Contrived specimens that have been generated from pooling of multiple samples or spiking of purified analyte to cover the measuring range may be used, but the contrived samples must be prepared to mimic clinical specimens as closely as possible. This testing must evaluate repeatability and reproducibility using a protocol from an FDA-recognized standard. (ii) Device performance data must be demonstrated through a clinical study and must include the following: (A) Data demonstrating clinical validity including the clinical sensitivity and specificity, and positive and negative predictive value of the test in the intended use population of patients with suspected mild traumatic brain injury ( *i.e.,* Glasgow Coma Score (GCS) of 13-15), or equivalent standard of care for determination of severity of traumatic brain injury (TBI).(B) Study must be performed using the operators and in settings that are representative of the types of operators and settings for which the device is intended to be used. (C) All eligible subjects must meet the well-defined study inclusion and exclusion criteria that define the intended use population. The prevalence of diseased or injured subjects in the study population must reflect the prevalence of the device's intended use population, or alternatively, statistical measures must be used to account for any bias due to enrichment of subpopulations of the intended use population. (D) All eligible subjects must have undergone a head computerized tomography (CT) scan or other appropriate clinical diagnostic standard used to determine the presence of an intracranial lesion as part of standard of care and must also be evaluated by the subject device. All clinical diagnostic standards used in the clinical study must follow standard clinical practice in the United States. (E) Relevant demographic variables and baseline characteristics including medical history and neurological history. In addition, head injury characteristics, neurological assessments, and physical evidence of trauma must be provided for each subject. This information includes but is not limited to the following: Time since head injury, time from head injury to CT scan, time from head injury to blood draw, GCS score or equivalent, experience of loss of consciousness, presence of confusion, episodes of vomiting, post-traumatic amnesia characteristics, presence of post-traumatic seizures, drug or alcohol intoxication, mechanism of injury, acute intracranial lesion type, neurosurgical lesion, and cranial fracture. (F) Each CT scan or other imaging result must be independently evaluated in a blinded manner by at least two board-certified radiologists to determine whether it is positive or negative as defined by the presence or absence of acute intracranial lesions. This independent review must be conducted without access to test results of the device. Prior to conducting the review, the criteria and procedures to be followed for scoring the images must be established, including the mechanism for determining consensus. (G) All the clinical samples must be tested with the subject device blinded to the TBI status and the neurological-lesion-status of the subject. (H) Details on how missing values in data are handled must be provided. (I) For banked clinical samples, details on storage conditions and storage period must be provided. In addition, a specimen stability study must be conducted for the duration of storage to demonstrate integrity of archived clinical samples. The samples evaluated in the assay test development must not be used to establish the clinical validity of the assays. (iii) Performance testing of device analytical specificity must include the most commonly reported concomitant medications present in specimens from the intended use population. Additionally, potential cross-reacting endogenous analytes must be evaluated at the highest concentration reported in specimens from the intended use population. (iv) Expected/reference values generated by testing a statistically appropriate number of samples from apparently healthy normal individuals. (2) The 21 CFR 809.10(a) and (b) compliant labeling must include the following limitations: (i) A limiting statement that this device is not intended to be used a stand-alone device but as an adjunct to other clinical information to aid in the evaluation of patients who are being considered for standard of care neuroimaging. (ii) A limiting statement that reads “A negative result is generally associated with the absence of acute intracranial lesions. An appropriate neuroimaging method is required for diagnosis of acute intracranial lesions.” (iii) As applicable, a limiting statement that reads “This device is for use by laboratory professionals in a clinical laboratory setting.”

Predicate Devices

• Banyan Brain Trauma Indicator (BTI) (DEN170045)

Submission Summary (Full Text)

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May 24, 2022

Abbott Laboratories Brian Ma Sr. Regulatory Affairs Specialist 400 College Road East Princeton, New Jersey 08540

Re: K201778

Trade/Device Name: i-STAT TBI Plasma cartridge with the i-STAT Alinity System Regulation Number: 21 CFR 866.5830 Regulation Name: Brain trauma assessment test Regulatory Class: Class II Product Code: QAT

Dear Brian Ma:

The Food and Drug Administration (FDA) is sending this letter to notify you of an administrative change related to your previous substantial equivalence (SE) determination letter for your device cleared on January 8, 2021. Specifically, FDA is updating this SE Letter due to a typographical error in the clearance date, which was incorrectly dated January 8, 2020.

Please note that the 510(k) submission was not re-reviewed. For questions regarding this letter please contact Ying (Katelin) Mao, OHT7: Office of In Vitro Diagnostics and Radiological Health, 301-796-6635, ying.mao(@)fda.hhs.gov .

Sincerely,

Ying Mao-S

Ying (Katelin) Mao, Ph.D. Chief Division of Immunology and Hematology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

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January 8, 2020

Abbott Laboratories Brian Ma Sr. Regulatory Affairs Specialist 400 College Road East Princeton, New Jersey 08540

Re: K201778

Trade/Device Name: i-STAT TBI Plasma cartridge with the i-STAT Alinity System Regulation Number: 21 CFR 866.5830 Regulation Name: Brain Trauma Assessment Test Regulatory Class: Class II Product Code: OAT Dated: December 4, 2020 Received: December 7, 2020

Dear Brian Ma:

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

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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 ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely,

Leonthena R. Carrington -S

Lea Carrington Director Division of Immunology and Hematology 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) K201778

#### Device Name

i-STAT TBI Plasma cartridge with the i-STAT Alinity System

#### Indications for Use (Describe)

The i-STAT TBI Plasma test is a panel of in vitro diagnostic immunoassays for the quantitative measurements of glial fibrillary acidic protein (GFAP) and ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) in plasma and a semiquantitative interpretation of test results derived from these measurements, using the i-STAT Alinity Instrument. The interpretation of test results is used, in conjunction with other clinical information, to aid in the evaluation of patients, 18 years of age or older, presenting with suspected mild traumatic brain injury (Glasgow Coma Scale score 13-15) within 12 hours of injury, to assist in determining the need for a CT (computed tomography) scan of the head. A 'Not Elevated' test interpretation is associated with the absence of acute traumatic intracranial lesions visualized on a head CT scan.

The test is to be used with plasma prepared from EDTA anticoagulated specimens in clinical laboratory settings by a healthcare professional. The i-STAT TBI Plasma test is not intended to be used in point of care settings.

| Type of Use (Select one or both, as applicable) |
|-------------------------------------------------|
|-------------------------------------------------|

X Prescription Use (Part 21 CFR 801 Subpart D)

| Over-The-Counter Use (21 CFR 801 Subpart C)

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# 510(k) Summary

The information in this 510(k) summary is being submitted in accordance with the requirements of 21 CFR 807.92.

| 1. | Submitter Information | | |
|----|-----------------------|--------------------------------------|--|
| | Owner | Abbott Point of Care Inc. | |
| | | 400 College Road East | |
| | | Princeton, NJ 08540 | |
| | Contact | Primary: Brian Ma, PhD | |
| | | Senior Specialist Regulatory Affairs | |
| | | Phone: [phone] | |
| | | Secondary: Susan Tibedo | |
| | | Director Regulatory Affairs | |
| | | Phone: [phone] | |

Date Prepared

January 6, 2021

## 2. Device Information

Proprietary Name i-STAT TBI Plasma Cartridge with the i-STAT Alinity System Glial fibrillary acidic protein (GFAP) Common Name Ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1)

| Product<br>code | Device Classification<br>name | Regulation<br>Number | Class | Panel |
|-----------------|---------------------------------|----------------------|--------------------------|------------|
| QAT | Brain trauma assessment<br>test | 866.5830 | II (Special<br>controls) | Immunology |

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#### 3. Predicate Device

Proprietary Name Banyan Brain Trauma Indicator (BTI)

510(k) Number DEN170045

| Product<br>code | Device Classification<br>name | Regulation<br>Number | Class | Panel |
|-----------------|---------------------------------|----------------------|-----------------------------|--------------------|
| QAT | Brain trauma<br>assessment test | 866.5830 | II<br>(Special<br>controls) | Immunology<br>(82) |

## 4. Device Description

The i-STAT TBI Plasma cartridge is a multiplex immunoassay that contains assays for both ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) and glial fibrillary acidic protein (GFAP). The assays test for the presence of these biomarkers in a plasma sample and yield a semi-quantitative test interpretation based on measurements of both UCH-L1 and GFAP in approximately 15 minutes. The i-STAT TBI Plasma cartridge is designed to be run only on the i-STAT Alinity instrument.

The i-STAT Alinity instrument is a handheld, in vitro diagnostic device designed to run only i-STAT test cartridges. The instrument is the main user interface of the i-STAT System and functions as the electro-mechanical interface to the test cartridge. The instrument executes the test cycle, acquires and processes the electrical sensor signals converting the signals into quantitative results. These functions are controlled by a microprocessor.

The i-STAT Alinity System is comprised of the i-STAT Alinity instrument, the i-STAT test cartridges and accessories (i-STAT Alinity Base Station, Electronic Simulator and Printer).

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#### 5. Intended Use Statement

The i-STAT TBI Plasma test is a panel of in vitro diagnostic immunoassays for the quantitative measurements of glial fibrillary acidic protein (GFAP) and ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) in plasma and a semiquantitative interpretation of test results derived from these measurements, using the i-STAT Alinity Instrument. The interpretation of test results is used, in conjunction with other clinical information, to aid in the evaluation of patients, 18 years of age or older, presenting with suspected mild traumatic brain injury (Glasgow Coma Scale score 13-15) within 12 hours of injury, to assist in determining the need for a CT (computed tomography) scan of the head. A 'Not Elevated' test interpretation is associated with the absence of acute traumatic intracranial lesions visualized on a head CT scan.

The test is to be used with plasma prepared from EDTA anticoagulated specimens in clinical laboratory settings by a healthcare professional. The i-STAT TBI Plasma test is not intended to be used in point of care settings.

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| Similarities and Differences: System (Test and Instrument) | | |
|------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Feature or<br>Characteristic | i-STAT TBI Plasma Cartridge<br>(Candidate) | Banyan BTI (DEN170045)<br>(Predicate) |
| Intended Use /<br>Indications for Use | The i-STAT TBI Plasma test is a<br>panel of in vitro diagnostic<br>immunoassays for the quantitative<br>measurements of glial fibrillary acidic<br>protein (GFAP) and ubiquitin<br>carboxyl-terminal hydrolase L1 (UCH-<br>L1) in plasma and a semi-quantitative<br>interpretation of test results derived<br>from these measurements, using the<br>i-STAT Alinity Instrument. The<br>interpretation of test results is used, in<br>conjunction with other clinical<br>information, to aid in the evaluation of<br>patients, 18 years of age or older,<br>presenting with suspected mild<br>traumatic brain injury (Glasgow Coma<br>Scale score 13-15) within 12 hours of<br>injury, to assist in determining the<br>need for a CT (computed<br>tomography) scan of the head. A 'Not<br>Elevated' test interpretation is<br>associated with the absence of acute<br>traumatic intracranial lesions<br>visualized on a head CT scan.<br>The test is to be used with plasma<br>prepared from EDTA anticoagulated<br>specimens in clinical laboratory<br>settings by a healthcare professional. | The Banyan BTI is an in vitro diagnostic<br>chemiluminescent enzyme-linked<br>immunosorbent assay (ELISA). The assay<br>provides a semi-quantitative measurement<br>of the concentrations of ubiquitin<br>C-terminal hydrolase-L1 (UCH-L1) and<br>glial fibrillary acidic protein (GFAP) in<br>human serum and is used with the<br>Synergy 2 Multi-mode Reader.<br>The assay results obtained from serum<br>collected within 12 hours of suspected<br>head injury are used, along with other<br>available clinical information, to aid in the<br>evaluation of patients 18 years of age and<br>older with suspected traumatic brain injury<br>(Glasgow Coma Scale score 13-15). A<br>negative assay result is associated with<br>the absence of acute intracranial lesions<br>visualized on a head CT (computed<br>tomography) scan. |
| Intended User | Clinical Laboratory | Clinical Laboratory |
| Measurands | GFAP and UCH-L1 | GFAP and UCH-L1 |
| Assay Technology | Enzyme-linked immunosorbent assay | Enzyme-linked immunosorbent assay |
| Assay Format | Single use multiplex cartridge (both<br>assays (GFAP and UCH-L1) in one<br>cartridge) | Two test kits on separate 96-well microtiter<br>plates - one for GFAP and one for UCH-<br>L1 |
| Detection<br>Technology | Electrochemical | Chemiluminescence |
| Specimen Type | Plasma | Serum |
| Sample Volume | 20 µL | GFAP kit: 150 µL<br>UCH-L1 kit: 100 µL |
| Similarities and Differences: System (Test and Instrument) | | |
| Feature or<br>Characteristic | i-STAT TBI Plasma Cartridge<br>(Candidate) | Banyan BTI (DEN170045)<br>(Predicate) |
| Preparation | Ready to Use | Manual preparation of reagents and plate. |
| Automation | Test and wash cycles are fully<br>automated after sample loading step | Manual sample loading, reagent mixing, as<br>well as transfer from incubation and<br>washing to read steps |
| Analytical<br>Measuring Interval | GFAP: 30 – 10,000 pg/mL<br>UCH-L1: 200 – 3,200 pg/mL | GFAP: 10 – 320 pg/mL<br>UCH-L1: 80 – 2,560 pg/mL |
| Time to Result | ~ 15 minutes | ~ 4 hours |
| Reportable Result | Quantitative results for GFAP and<br>UCH-L1 and semi-quantitative<br>interpretation | Quantitative results for GFAP and UCH-L1<br>and semi-quantitative interpretation |
| Instrument Platform | i-STAT Alinity | Synergy 2 Multi-Mode Microplate Reader<br>(BioTek Instruments, Inc.) |
| Controls | GFAP and UCH-L1 combined: 2<br>levels (Control 1, Control 2) | GFAP: 2 levels (Control 1, Control 2)<br>UCH-L1: 2 levels (Control 1, Control 2) |
| Calibration | No calibration needed by the end<br>user, calibration is pre-set during<br>manufacture of the cartridge | Calibration curve generated by end user<br>for each run using six standards |

# 6. Summary Comparison of Technological Characteristics

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## 7. Performance Characteristics

## 1. Analytical Performance

## a. Precision/Reproducibility

Semi-quantitative precision: The precision of the GFAP and UCH-L1 assays in the i-STAT TBI plasma cartridge with the i-STAT Alinity System was evaluated using plasma samples representing nine (9) levels of GFAP and seven (7) levels of UCH-L1 spanning the reportable range as well as the i-STAT TBI Controls (L1 and L2). This single-site study was based on CLSI document EP05-A3: Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline-Third Edition. Each sample was tested for at least 20 days with two (2) runs per day and two (2) results per run for a total of 80 measurements per sample per cartridge lot. Runs were separated by a minimum of 2 hours. The plasma samples were made of pooled plasma from healthy normal donors spiked with recombinant GFAP and UCH-L1 antigens or spiked with

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native antigens from pooled TBI patient plasma as shown in the tables below. The components of variability were estimated for GFAP and UCH-L1 and the precision results are shown in Table 1, Table 2, and Table 3.

| Table 1: Estimate of GFAP Assay Precision | | | | | | | | | | | | |
|-------------------------------------------|-------|-----------------|---------------|-----------|---------------|-----------|---------------|-----------|---------------|-----------|-----------------------|-----------|
| Sample | N | Mean<br>(pg/mL) | Repeatability | | Between-Run | | Between-Day | | Between-Lot | | Within-<br>Laboratory | |
| | | | SD<br>(pg/mL) | CV<br>(%) | SD<br>(pg/mL) | CV<br>(%) | SD<br>(pg/mL) | CV<br>(%) | SD<br>(pg/mL) | CV<br>(%) | SD<br>(pg/mL) | CV<br>(%) |
| 1 | 238‡ | 17.0 | 1.76 | 10.4% | 0.91 | 5.4% | 0.61 | 3.6% | 1.31 | 7.7% | 2.46 | 14.5% |
| 2 | 238‡ | 30.8 | 2.49 | 8.1% | 0.00 | 0.0% | 0.00 | 0.0% | 0.52 | 1.7% | 2.55 | 8.3% |
| 3 | 238‡ | 65.6 | 3.21 | 4.9% | 0.87 | 1.3% | 1.03 | 1.6% | 0.62 | 0.9% | 3.54 | 5.4% |
| 4 | 238*§ | 104.9 | 3.37 | 3.2% | 2.08 | 2.0% | 0.00 | 0.0% | 1.50 | 1.4% | 4.24 | 4.0% |
| 5 | 238‡ | 962.9 | 22.42 | 2.3% | 13.61 | 1.4% | 17.33 | 1.8% | 21.17 | 2.2% | 37.90 | 3.9% |
| 6 | 160 | 2029.5 | 39.18 | 1.9% | 26.30 | 1.3% | 19.10 | 0.9% | 94.89 | 4.7% | 107.69 | 5.3% |
| 7 | 240 | 3139.5 | 75.98 | 2.4% | 35.92 | 1.1% | 49.34 | 1.6% | 97.09 | 3.1% | 137.57 | 4.4% |
| 8 | 160*† | 5713.3 | 143.96 | 2.5% | 42.68 | 0.7% | 65.72 | 1.2% | 170.29 | 3.0% | 236.36 | 4.1% |
| 9 | 159† | 7537.2 | 129.57 | 1.7% | 133.30 | 1.8% | 35.89 | 0.5% | 187.57 | 2.5% | 266.51 | 3.5% |

**Table 1: Estimate of GFAP Assay Precision**

*Additional GFAP result(s) was obtained due to cartridge re-run due to a UCH-L1 star-out

†one (1) outlier removed from analysis

キtwo (2) outliers removed from analysis

Sthree (3) outliers removed from analysis

| Table 2: Estimate of UCH-L1 Assay Precision | | | | | | | | | | | | |
|---------------------------------------------|------|-----------------|---------------|-----------|---------------|-----------|---------------|-----------|---------------|-----------|-----------------------|-----------|
| | | | Repeatability | | Between-Run | | Between-Day | | Between-Lot | | Within-<br>Laboratory | |
| Sample | N | Mean<br>(pg/mL) | SD<br>(pg/mL) | CV<br>(%) | SD<br>(pg/mL) | CV<br>(%) | SD<br>(pg/mL) | CV<br>(%) | SD<br>(pg/mL) | CV<br>(%) | SD<br>(pg/mL) | CV<br>(%) |
| 1 | 238‡ | 72.5 | 4.88 | 6.7% | 1.73 | 2.4% | 0.00 | 0.0% | 3.93 | 5.4% | 6.50 | 9.0% |
| 2 | 239† | 300.1 | 15.12 | 5.0% | 5.94 | 2.0% | 0.00 | 0.0% | 15.54 | 5.2% | 22.48 | 7.5% |
| 3 | 240 | 519.9 | 29.56 | 5.7% | 1.54 | 0.3% | 13.38 | 2.6% | 8.21 | 1.6% | 33.51 | 6.4% |
| 4 | 238‡ | 1058.9 | 56.88 | 5.4% | 22.59 | 2.1% | 15.13 | 1.4% | 33.6 | 3.2% | 71.44 | 6.7% |
| 5 | 159* | 1639.6 | 91.57 | 5.6% | 8.72 | 0.5% | 15.74 | 1.0% | 28.46 | 1.7% | 97.56 | 6.0% |
| 6 | 240 | 2067.4 | 111.09 | 5.4% | 54.99 | 2.7% | 46.01 | 2.2% | 15.00 | 0.7% | 133.06 | 6.4% |
| 7 | 239† | 2849.7 | 145.4 | 5.1% | 100.56 | 3.5% | 0.00 | 0.0% | 15.16 | 0.5% | 177.44 | 6.2% |

*one (1) result was unavailable due to star-out

*one (1) outlier removed from analysis

‡two (2) outliers removed from analysis

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| Table 3: Estimate of GFAP and UCH-L1 Assay Precision with i-STAT TBI Controls | | | | | | | | | | | | |
|-------------------------------------------------------------------------------|-------|-----------------|---------------|-----------|---------------|-----------|---------------|-----------|---------------|-----------|-----------------------|-----------|
| Sample | N | Mean<br>(pg/mL) | Repeatability | | Between-Run | | Between-Day | | Between-Lot** | | Within-<br>Laboratory | |
| | | | SD<br>(pg/mL) | CV<br>(%) | SD<br>(pg/mL) | CV<br>(%) | SD<br>(pg/mL) | CV<br>(%) | SD<br>(pg/mL) | CV<br>(%) | SD<br>(pg/mL) | CV<br>(%) |
| GFAP Assay | | | | | | | | | | | | |
| L1 | 238*§ | 196.7 | 9.94 | 5.1% | 2.69 | 1.4% | 2.25 | 1.1% | 5.70 | 2.9% | 11.98 | 6.1% |
| L2 | 242* | 5153.8 | 236.89 | 4.6% | 94.93 | 1.8% | 28.10 | 0.5% | 183.00 | 3.6% | 315.29 | 6.1% |
| UCH-L1 Assay | | | | | | | | | | | | |
| L1 | 239† | 562.6 | 29.79 | 5.3% | 9.57 | 1.7% | 11.92 | 2.1% | 13.21 | 2.3% | 36.00 | 6.4% |
| L2 | 240 | 1624.7 | 90.14 | 5.5% | 53.68 | 3.3% | 0.00 | 0.0% | 32.25 | 2.0% | 109.76 | 6.8% |

* Additional GFAP result was obtained due to cartridge re-run due to a UCH-L1 star-out

*one (1) outlier removed from analysis

‡two (2) outliers removed from analysis

§three (3) outliers removed from analysis

**This refers to precision estimates calculated between cartridge lots. A single lot of i-STAT TBI controls was used for this study.

Qualitative precision: The qualitative agreement of cartridge results relative to the expected sample result (mean) was evaluated for the 80 measurements per sample per cartridge lot for each assay from the semi-quantitative analysis above. The mean, total number of replicates, total number of elevated results, and % correct call for each sample level are presented in Table 4 for GFAP and Table 5 for UCH-L1.

| Table 4: GFAP assay results for qualitative precision analysis | | | | | |
|----------------------------------------------------------------|-----|-----------------|-------------------------------|-------------------------------------------------------|-------------------|
| Plasma<br>Sample | Lot | Mean<br>(pg/mL) | Total Number<br>of Replicates | Total Number of<br>Results at or<br>above the cut-off | % Correct<br>Call |
| 1A | A | 18.5 | 80 | 0 | 100% |
| | B | 15.6 | 80 | 0 | 100% |
| | C | 17.1 | 80 | 0 | 100% |
| 2B | D | 30.9 | 80 | 59 | 74% |
| | E | 30.5 | 80 | 50 | 63% |
| | F | 31.5 | 80 | 69 | 86% |
| 3C | D | 65.7 | 80 | 80 | 100% |
| | E | 64.8 | 80 | 80 | 100% |
| | F | 66.0 | 80 | 80 | 100% |
| 4C | A | 106.3 | 80 | 80 | 100% |
| | B | 102.7 | 80 | 80 | 100% |
| | C | 105.1 | 81 | 81 | 100% |
| 5C | A | 939.3 | 80 | 80 | 100% |
| | B | 979.6 | 80 | 80 | 100% |
| | C | 973.8 | 80 | 80 | 100% |
| 6C | G | 2096.8 | 80 | 80 | 100% |
| | H | 1962.2 | 80 | 80 | 100% |
| 7C | A | 3050.2 | 80 | 80 | 100% |

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| Table 4: GFAP assay results for qualitative precision analysis | | | | | |
|----------------------------------------------------------------|-----|-----------------|-------------------------------|-------------------------------------------------------|-------------------|
| Plasma<br>Sample | Lot | Mean<br>(pg/mL) | Total Number<br>of Replicates | Total Number of<br>Results at or<br>above the cut-off | % Correct<br>Call |
| | B | 3244.7 | 80 | 80 | 100% |
| | C | 3123.6 | 80 | 80 | 100% |
| 8C | G | 5579.9 | 80 | 80 | 100% |
| | H | 5832.8 | 83 | 83 | 100% |
| 9c | G | 7404.2 | 80 | 80 | 100% |
| | H | 7685.7 | 80 | 80 | 100% |
| i-STAT TBI Controls | | | | | |
| | A | 203.4 | 81 | 81 | 100% |
| L1C | B | 193.3 | 80 | 80 | 100% |
| | C | 195.1 | 80 | 80 | 100% |
| L2C | A | 5162.0 | 82 | 82 | 100% |
| | B | 5335.2 | 80 | 80 | 100% |
| | C | 4964.2 | 80 | 80 | 100% |

A Below cut-off

^ Below cut-off
B Near cut-off (mean ± 25%)
© Above cut-off

^c Above cut-off

| Table 5: UCH-L1 assay results for qualitative precision analysis | | | | | |
|------------------------------------------------------------------|-----|-----------------|-------------------------------|-------------------------------------------------------|-------------------|
| Plasma<br>Sample | Lot | Mean<br>(pg/mL) | Total Number<br>of Replicates | Total Number of<br>Results at or<br>above the cut-off | % Correct<br>Call |
| 1A | D | 70.7 | 80 | 0 | 100% |
| | E | 73.5 | 80 | 0 | 100% |
| | F | 77.1 | 80 | 0 | 100% |
| 2B | D | 295.6 | 80 | 0 | 100% |
| | E | 289.0 | 80 | 1 | 99% |
| | F | 317.5 | 80 | 0 | 100% |
| 3C | A | 525.1 | 80 | 80 | 100% |
| | B | 509.1 | 80 | 80 | 100% |
| | C | 525.6 | 80 | 80 | 100% |
| 4C | A | 1039.8 | 80 | 80 | 100% |
| | B | 1039.0 | 80 | 80 | 100% |
| | C | 1098.8 | 80 | 80 | 100% |
| 5C | G | 1617.9 | 79 | 79 | 100% |
| | H | 1661.0 | 80 | 80 | 100% |
| 6C | A | 2078.7 | 80 | 80 | 100% |
| | B | 2040.2 | 80 | 80 | 100% |
| | C | 2083.2 | 80 | 80 | 100% |
| 7C | A | 2865.0 | 80 | 80 | 100% |
| | B | 2819.5 | 80 | 80 | 100% |
| | C | 2855.7 | 80 | 80 | 100% |
| i-STAT TBI Controls | | | | | |
| L1C | A | 556.8 | 80 | 80 | 100% |
| | B | 575.2 | 80 | 80 | 100% |
| | C | 552.6 | 80 | 80 | 100% |
| L2C | A | 1601.2 | 80 | 80 | 100% |
| | B | 1664.7 | 80 | 80 | 100% |

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| Table 5: UCH-L1 assay results for qualitative precision analysis | | | | | |
|------------------------------------------------------------------|-----|-----------------|-------------------------------|-------------------------------------------------------|-------------------|
| Plasma<br>Sample | Lot | Mean<br>(pg/mL) | Total Number<br>of Replicates | Total Number of<br>Results at or<br>above the cut-off | % Correct<br>Call |
| | C | 1608.1 | 80 | 80 | 100% |

A Below cut-off

B Near cut-off (mean ± 25%)

C Above cut-off

Semi-quantitative multi-site precision: The precision performance of the GFAP and UCH-L1 assays in the i-STAT TBI Plasma cartridge on the i-STAT Alinity System was evaluated using seven (7) test materials representing six (6) levels of GFAP and five (5) levels of UCH-L1 at three (3) external clinical sites. At each site, each test material was tested once per day for five (5) days by two (2) different operators, with each operator using three (3) i-STAT Alinity Instruments. The plasma samples were made of pooled plasma from healthy normal donors spiked with recombinant GFAP and UCH-L1 antigens or spiked with native antigens from pooled TBI patient plasma as shown in the tables below. Thirty (30) replicates were obtained for the applicable assay analyzed for each test material at each of the three (3) sites for a total of 90 replicates for each test material. The estimates of GFAP and UCH-L1 precision are shown in Table 6 and Table 7.

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| | | | ble 6: i-STAT TBI Plasma Multi-site Precision - GFAP (pg/mL) - 2 Test - - - - - - - - Moan | | | nL) - All Sites | | | | | | |…