Diazyme Fibrinogen Assay, Diazyme Fibrinogen Calibrator Set, Diazyme Fibrinogen Control Set

K143470 · Diazyme Laboratories · GIS · Jan 31, 2016 · Hematology

Device Facts

• Record ID: K143470
• Device Name: Diazyme Fibrinogen Assay, Diazyme Fibrinogen Calibrator Set, Diazyme Fibrinogen Control Set
• Applicant: Diazyme Laboratories
• Product Code: GIS · Hematology
• Decision Date: Jan 31, 2016
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 864.7340
• Device Class: Class 2

Indications for Use

The Diazyme Fibrinogen Assay is for the quantitative determination of fibrinogen levels in citrated human plasma. For in vitro diagnostic use only. The Diazyne Fibringen Calibrator Set is intended for use in the Diazyme Fibrinogen Assay. For in vitro diagnostic use only. The Diazyme Fibringen Control Set is intended for use as quality controls for the Diazyme Fibrinogen Assay. For in vitro diagnostic use only.

Device Story

Diazyme Fibrinogen Assay is an immunoturbidimetric assay for measuring fibrinogen in citrated human plasma. Input: citrated plasma sample. Principle: fibrinogen binds to anti-fibrinogen antibody to form immune complexes; turbidity measured optically at 340nm. Output: fibrinogen concentration calculated via 4-point calibration curve interpolation. Used in clinical laboratories; operated by trained professionals on Roche Modular P analyzer. Analyzer automates dilution, reagent/sample handling, and absorbance measurement. Healthcare providers use results to assess coagulation status, inflammation, or fibrinolysis. Benefits: aids diagnosis of inherited/acquired fibrinogen disorders, DIC, and monitoring of inflammatory states.

Clinical Evidence

Bench testing only. Precision/reproducibility evaluated per CLSI EP5-A2 (CV ≤ 8% within-run, ≤ 10% total). Linearity confirmed up to 1032.8 mg/dL per CLSI EP6-A. Method comparison (N=176) against predicate showed slope 0.986, R² 0.9946. Interference testing (CLSI EP7-A2) showed no significant interference from endogenous/exogenous substances. No clinical data.

Technological Characteristics

Immunoturbidimetric assay; measures turbidity at 340 nm. Reagents contain anti-human fibrinogen Ig fraction. Calibrators/controls are lyophilized, serum-based. Designed for use on Roche Modular P analyzer. Traceable to WHO reference material NIBSC 09/242. Software parameters programmed into analyzer.

Indications for Use

Indicated for quantitative determination of fibrinogen levels in citrated human plasma. Performance not established in neonate or pediatric populations.

Regulatory Classification

Identification

A fibrinogen determination system is a device that consists of the instruments, reagents, standards, and controls used to determine the fibrinogen levels in disseminated intravascular coagulation (nonlocalized clotting within the blood vessels) and primary fibrinolysis (the dissolution of fibrin in a blood clot).

Special Controls

*Classification.* Class II (special controls). A control or fibrinogen standard intended for use with a fibrinogen determination system is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 864.9.

Predicate Devices

• Kamiya K-Assay Fibrinogen (K993482)

Submission Summary (Full Text)

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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

A. 510(k) Number:
K143470

B. Purpose for Submission:
New device

C. Measurand:
Fibrinogen

D. Type of Test:
Quantitative immunoturbidimetric assay for fibrinogen

E. Applicant:
Diazyme Laboratories

F. Proprietary and Established Names:
Diazyme Fibrinogen Assay
Diazyme Fibrinogen Calibrator Set
Diazyme Fibrinogen Control Set

G. Regulatory Information:
1. Regulation section:
21CFR 864.7340, Fibrinogen determination system
2. Classification:
Class II
3. Product code:
GIS, Test, fibrinogen
GFX, Fibrinogen standard

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GIL, Plasma, Fibrinogen Control

4. Panel:
81 Hematology

H. Intended Use:

1. Intended use(s):
The Diazyme Fibrinogen Assay is for the quantitative determination of fibrinogen levels in citrated human plasma. For in vitro diagnostic use only.
The Diazyme Fibrinogen Calibrator Set is intended for use in the calibration of the Diazyme Fibrinogen Assay. For in vitro diagnostic use only.
The Diazyme Fibrinogen Control Set is intended for use as quality controls for the Diazyme Fibrinogen Assay. For in vitro diagnostic use only.
The performance of this device has not been established in neonate and pediatric patient populations.

2. Indication(s) for use:
Same as intended use

3. Special conditions for use statement(s):
For in vitro diagnostic use only

4. Special instrument requirements:
The Diazyme Fibrinogen Assay is to be used with the Roche Modular P analyzer.

I. Device Description:

Diazyme Fibrinogen Assay System consists of the following:

1. The Diazyme Fibrinogen Assay contains reagent R1 and reagent R2. Reagent 1 contains buffer, saline, and sodium azide. Reagent 2 contains anti-human fibrinogen Ig fraction tittered to optimal concentrations dependent on sera lot and sodium azide.

2. Diazyme Fibrinogen Calibrator Set contains three levels of serum based calibrator. The calibrators are in lyophilized form containing purified human fibrinogen antigen, defibrinated human serum and sodium azide.

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3. Diazyme Fibrinogen Control Set contains two levels of serum based control. The controls are in lyophilized form containing purified human fibrinogen antigen, defibrinated human serum and sodium azide.

The Diazyme Fibrinogen Assay is run on Roche Modular P analyzer. The patient's venous blood sample is first collected by a phlebotomist and then submitted to the clinical laboratory for determination of fibrinogen concentration using Diazyme Fibrinogen Assay. The Diazyme Fibrinogen Modular P application parameters are programmed into the Modular P analyzer. The reagents, calibrators and controls are loaded into the analyzer. Reagent and sample pipettes automatically aspirate and dispense specified amounts of reagent or calibrators, controls, and patient sample into reaction cells. The change in absorbance is measured at specified wavelengths.

The Roche Modular P analyzer calculates the fibrinogen concentration of a patient sample by interpolation of the obtained signal to a stored 4-point calibration curve.

## J. Substantial Equivalence Information:

1. Predicate device name(s):

K-ASSAY Fibrinogen

2. Predicate 510(k) number(s):

K993482

3. Comparison with predicate:

| Similarities | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Indications for Use | The Diazyme Fibrinogen Assay is for the quantitative determination of fibrinogen levels in citrated human plasma. For in vitro diagnostic use only. | The K-ASSAY Fibrinogen Assay is intended to be used to determine the fibrinogen levels in citrated human plasma. For in vitro diagnostic use only. |
| Test Principle | Immunoturbidimetric | Same |
| Type of Test | Quantitative | Same |
| Specimen Type | Plasma | Same |
| Assay Range | 100 - 900 mg/dL | Same |
| Reagent 2 | Goat anti-human fibrinogen antiserum | Same |
| Calibrator Set | Lyophilized | Same |
| Control Set | Lyophilized | Same |

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| Differences | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Reagent 1 | Phosphate Buffered Saline | Tris Buffered Saline |
| Calibrator Set | Three levels in 1 x 1.0 mL vials | Single-level in 1 x 1.0 mL vials |
| Control Set | Serum based
Two levels in 1 x 1.0 mL vials | Plasma based
Two levels in 5 x 1.0 mL vials |

## K. Standard/Guidance Document Referenced (if applicable):

CLSI-EP5: Evaluation of Precision Performance of Quantitative Measurement Methods

CLSI-EP6: Evaluation of the Linearity of Quantitative Analytical Methods

CLSI-EP7: Interference Testing in Clinical Chemistry

CLSI-EP9: Measurement Procedure Comparison and Bias Estimation using Patient Samples

CLSI-EP17: Protocols for Determination of Limits of Detection and Limits of Quantitation

CLSI-EP28: Defining, Establishing and Verifying Reference Intervals in the Clinical Laboratories

## L. Test Principle:

The Diazyme Fibrinogen Assay is based on an immunoturbidimetric test principle. Fibrinogen in plasma binds to specific anti-fibrinogen antibody and forms immune complexes. The degree of turbidity caused by immune complexes can be measured optically (340 nm), and is proportional to the quantity of fibrinogen in the sample. The Roche Modular P analyzer calculates the fibrinogen concentration by interpolation of the obtained signal of a 4-point calibration curve prepared from calibrators of known concentrations.

## M. Performance Characteristics:

1. Analytical performance:

a. Precision/Reproducibility:

**Precision Study (Reagent, Calibrator and Control):**

The precision of the Diazyme Fibrinogen Assay, Calibrator and Control were evaluated according to CLSI EP5-A2 guideline on the Roche Modular P analyzer.

For reagent repeatability study, three lots of the reagents were used. For each lot of reagents, the same set of six levels of human citrated plasma specimens containing 100-115 mg/dL, 200-215 mg/dL, 300-340 mg/dL, 400-460 mg/dL, 700-720 mg/dL, and 800-860 mg/dL of fibrinogen, respectively, were tested. Each sample run was

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tested in duplicate, 2 runs per day for 20 working days according to CLSI EP5-A2. Within-run, between-run, between-lot, between-day, and total precision within site were calculated. The pre-established acceptance criterion, $\mathrm{CV} \leq 8\%$ , was met.

| Reagent Repeatability Studies (3 lots) | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Sample | N | Mean mg/dL | Within-Run (SD, %CV) | Between-Run (SD, | Between-Lot | Between-Day | Total (SD, %CV) |
| Plasma 1 | 240 | 453.4 | 8.0, 1.8% | 9.4, 2.1% | 16.2, 3.6% | 10.6, 2.3% | 23.0, 5.1% |
| Plasma 2 | 240 | 333.4 | 5.8, 1.7% | 5.2, 1.6% | 10.7, 3.2% | 7.4, 2.2% | 15.1, 4.5% |
| Plasma 3 | 240 | 208.6 | 3.0, 1.4% | 3.2, 1.5% | 6.6, 3.2% | 4.9, 2.4% | 9.3, 4.5% |
| Plasma 4 | 240 | 107.3 | 1.6, 1.5% | 1.7, 1.6% | 3.6, 3.4% | 2.8, 2.6% | 5.2, 4.8% |
| Plasma 5 | 240 | 706.8 | 11.9, 1.7% | 13.9, 2.0% | 27.1, 3.8% | 20.1, 2.8% | 38.4, 5.4% |
| Plasma 6 | 240 | 823.9 | 16.3, 2.0% | 12.7, 1.5% | 30.3, 3.7% | 22.3, 2.7% | 42.9, 5.2% |

For production calibrator and control repeatability studies, three lots of the calibrator (3 levels) and one lots of control (2 levels) were tested according to CLSI EP5-A2 guideline. Within-run, between-run, between-lot, between-day, and total precision within site were calculated. The pre-established acceptance criterion, $\mathrm{CV} \leq 8\%$ , was met.

| Calibrator Repeatability Studies (3 lots) | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Sample | N | Mean mg/dL | Within-Run (SD, %CV) | Between-Run (SD, %CV) | Between-Lot (SD,%CV) | Between-Day (SD,%CV) | Total (SD, %CV) |
| Level 1 | 360 | 262.0 | 5.7, 2.2% | 0, 0% | 13.3, 5.1% | 12.1, 4.6% | 18.8, 7.2% |
| Level 2 | 360 | 578.0 | 17.9, 3.1% | 0, 0% | 27.9, 4.8% | 22.5, 3.9% | 40.1, 6.9% |
| Level 3 | 360 | 1130.1 | 21.2, 1.9% | 0, 0% | 44.9, 4.0% | 40.4, 3.6% | 64.0, 5.7% |
| Control Repeatability Studies (1 lot of Control and 2 lots of Reagent) | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Sample | N | Mean mg/dL | Within-Run (SD, %CV) | Between-Run (SD, %CV) | Between-Lot (SD,%CV) | Between-Day (SD,%CV) | Total (SD, %CV) |
| Level 1 | 160 | 173.2 | 2.3, 1.3% | 1.4, 0.8% | 4.1, 2.4% | 3.1, 1.8% | 4.1, 2.4% |
| Level 2 | 160 | 607.5 | 8.2, 1.4% | 6.5, 1.1% | 15.1, 2.5% | 11.1, 1.8% | 15.2, 2.5% |

# Reproducibility Study (Reagent, Calibrator and Control):

A multi-site precision study was performed at two external sites and one internal site on the Modular P analyzer. Six citrated plasma samples, containing one sample close to the very low end of the analytical measuring range (AMR) and one sample close to the very high end of the AMR, were tested in duplicates per run, two runs per day for five nonconsecutive working days. One lot of reagents was tested across three testing sites by three operators on three Modular P analyzers. Additionally, three lots of calibrators and three lots of controls were tested in duplicates per run, two runs per day for five nonconsecutive working days with one lot of reagents at three sites by three operators on three Modular P analyzers. Within-run, between-run, between-lot, between-day, between-site, and total precision were calculated. The pre-established

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acceptance criteria, $\mathrm{CV} \leq 8\%$ for within-run, between-run, between-lot, between day, within site and between-site and $\mathrm{CV} \leq 10\%$ for total, were met.

| Reproducibility Within-run Study (3 Sites) | | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Sample | Lots | N | Mean mg/dL | Site 1 | | Site 2 | | Site 3 | |
| | | | | SD | %CV | SD | %CV | SD | %CV |
| Plasma 1 | 1 | 20 | 441.1 | 11.4 | 2.8% | 9.33 | 2.5% | 3.7 | 0.8% |
| Plasma 2 | 1 | 20 | 323.9 | 6.6 | 2.2% | 5.29 | 1.9% | 5.3 | 1.6% |
| Plasma 3 | 1 | 20 | 205.4 | 3.6 | 1.8% | 1.9 | 1.0% | 2.7 | 1.3% |
| Plasma 4 | 1 | 20 | 107.4 | 0.8 | 0.8% | 1.55 | 1.6% | 1.2 | 1.2% |
| Plasma 5 | 1 | 20 | 709.5 | 22.9 | 3.2% | 26.47 | 3.9% | 7.9 | 1.1% |
| Plasma 6 | 1 | 20 | 822.7 | 9.2 | 1.1% | 11.0 | 1.3% | 18.7 | 2.3% |
| Calibrator 1 | 3 | 60 | 250.7 | 3.3 | 1.3% | 4.1 | 1.7% | 3.9 | 1.5% |
| Calibrator 2 | 3 | 60 | 502.1 | 12.6 | 2.5% | 6.6 | 1.3% | 12.8 | 2.5% |
| Calibrator 3 | 3 | 60 | 1077.3 | 17.3 | 1.7% | 16.7 | 1.6% | 17.2 | 1.5% |
| Control 1 | 3 | 60 | 179.4 | 2.1 | 1.2% | 2.5 | 1.4% | 4.6 | 2.5% |
| Control 2 | 3 | 60 | 609.6 | 14.6 | 2.4% | 15.3 | 2.5% | 8.6 | 1.4% |
| Reproducibility Between-run Study (3 Sites) | | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Sample | Lots | N | Mean mg/dL | Site 1 | | Site 2 | | Site 3 | |
| | | | | SD | %CV | SD | %CV | SD | %CV |
| Plasma 1 | 1 | 20 | 441.1 | 4.9 | 1.2% | 10.86 | 2.9% | 13.56 | 3.1% |
| Plasma 2 | 1 | 20 | 323.9 | 0.0 | 0.0% | 5.27 | 1.9% | 4.17 | 1.3% |
| Plasma 3 | 1 | 20 | 205.4 | 0.0 | 0.0% | 3.07 | 1.7% | 0.00 | 0.0% |
| Plasma 4 | 1 | 20 | 107.4 | 0.7 | 0.7% | 0.82 | 0.9% | 1.40 | 1.3% |
| Plasma 5 | 1 | 20 | 709.5 | 33.4 | 4.7% | 0.0 | 0.0% | 17.63 | 2.5% |
| Plasma 6 | 1 | 20 | 822.7 | 10.7 | 1.3% | 11.2 | 1.4% | 13.56 | 3.1% |
| Calibrator 1 | 3 | 60 | 250.7 | 3.2 | 1.3% | 9.9 | 4.1% | 1.2 | 0.5% |
| Calibrator 2 | 3 | 60 | 502.1 | 4.7 | 1.0% | 11.6 | 2.3% | 14.1 | 2.8% |
| Calibrator 3 | 3 | 60 | 1077.3 | 19.1 | 1.8% | 19.9 | 1.9% | 0 | 0% |
| Control 1 | 3 | 60 | 179.4 | 2.9 | 1.6% | 2.6 | 1.5% | 0 | 0% |
| Control 2 | 3 | 60 | 609.6 | 9.4 | 1.6% | 13.6 | 2.3% | 14.2 | 2.3% |
| Reproducibility Between-day Study (3 Sites) | | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Sample | Lots | N | Mean mg/dL | Site 1 | | Site 2 | | Site 3 | |
| | | | | SD | %CV | SD | %CV | SD | %CV |
| Plasma 1 | 1 | 20 | 441.1 | 9.5 | 2.4% | SD | %CV | 0.00 | 0.0% |
| Plasma 2 | 1 | 20 | 323.9 | 5.0 | 1.7% | 8.21 | 2.2% | 7.79 | 2.4% |
| Plasma 3 | 1 | 20 | 205.4 | 3.3 | 1.7% | 2.52 | 0.9% | 6.08 | 3.0% |
| Plasma 4 | 1 | 20 | 107.4 | 1.0 | 1.0% | 0.6 | 0.3% | 3.25 | 3.0% |
| Plasma 5 | 1 | 20 | 709.5 | 24.1 | 3.4% | 1.55 | 1.6% | 4.36 | 0.6% |
| Plasma 6 | 1 | 20 | 822.7 | 0.0 | 0.0% | 28.76 | 4.2% | 0.00 | 0.0% |
| Calibrator 1 | 3 | 60 | 250.7 | 7.3 | 3.0% | 2.5 | 1.0% | 10.7 | 4.1% |
| Calibrator 2 | 3 | 60 | 502.1 | 14.2 | 2.8% | 17.0 | 3.4% | 8.6 | 1.7% |
| Calibrator 3 | 3 | 60 | 1077.3 | 15.0 | 1.4% | 35.2 | 3.3% | 21.7 | 1.9% |
| Control 1 | 3 | 60 | 179.4 | 4.2 | 2.4% | 5.0 | 2.8% | 5.6 | 3.0% |
| Control 2 | 3 | 60 | 609.6 | 19.4 | 3.2% | 21.9 | 3.6% | 0.00 | 0.0% |

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| Reproducibility Within-site (3 Sites) | | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Sample | Lots | N | Mean mg/dL | Site 1 | | Site 2 | | Site 3 | |
| | | | | SD | %CV | SD | %CV | SD | %CV |
| Plasma 1 | 1 | 20 | 408.5 | 10.2 | 3% | 8.8 | 2% | 27.5 | 7% |
| Plasma 2 | 1 | 20 | 303.1 | 6.2 | 2% | 2.3 | 1% | 21.1 | 7% |
| Plasma 3 | 1 | 20 | 195.5 | 2.9 | 2% | 1.8 | 1% | 9.6 | 5% |
| Plasma 4 | 1 | 20 | 100.0 | 1.2 | 1% | 1.1 | 1% | 5.0 | 5% |
| Plasma 5 | 1 | 20 | 701.5 | 22.4 | 3% | 23.8 | 3% | 29.9 | 4% |
| Plasma 6 | 1 | 20 | 818.4 | 13.6 | 2% | 7.7 | 1% | 9.1 | 1% |
| Calibrator 1 | 3 | 60 | 250.7 | 3.8 | 2% | 6.0 | 2% | 11.7 | 5% |
| Calibrator 2 | 3 | 60 | 502.1 | 11.0 | 2% | 10.9 | 2% | 14.0 | 3% |
| Calibrator 3 | 3 | 60 | 1077.3 | 17.1 | 2% | 14.9 | 1% | 47.0 | 4% |
| Control 1 | 3 | 60 | 179.4 | 3.2 | 2% | 1.9 | 1% | 6.1 | 3% |
| Control 2 | 3 | 60 | 609.6 | 13.2 | 2% | 12.6 | 2% | 16.4 | 3% |
| Reproducibility Sites Combined and Total | | | | | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Sample | N | Mean mg/dL | With-run | | Between-run | | Between-day | | Between-Site | | Total | |
| | | | SD | %CV | SD | %CV | SD | %CV | SD | %CV | SD | %CV |
| Plasma 1 | 60 | 408.5 | 10.2 | 3% | 8.8 | 2% | 27.5 | 7% | 29.9 | 7.3% | 42.8 | 10 % |
| Plasma 2 | 60 | 303.1 | 6.2 | 2% | 2.3 | 1% | 21.1 | 7% | 21.6 | 7.1% | 30.9 | 10 % |
| Plasma 3 | 60 | 195.5 | 2.9 | 2% | 1.8 | 1% | 9.6 | 5% | 9.9 | 5.1% | 14.2 | 7% |
| Plasma 4 | 60 | 100.0 | 1.2 | 1% | 1.1 | 1% | 5.0 | 5% | 5.1 | 5.1% | 7.4 | 7% |
| Plasma 5 | 60 | 701.5 | 22.4 | 3% | 23.8 | 3% | 29.9 | 4% | 43.9 | 6.2% | 62.2 | 9% |
| Plasma 6 | 60 | 818.4 | 13.6 | 2% | 7.7 | 1% | 9.1 | 1% | 18.0 | 2.2% | 25.5 | 3% |
| Calibrator1 | 180 | 250.7 | 3.8 | 2% | 6.0 | 2% | 11.7 | 5% | 13.6 | 5.4% | 19.3 | 8% |
| Calibrator2 | 180 | 502.1 | 11.0 | 2% | 10.9 | 2% | 14.0 | 3% | 20.8 | 4.1% | 29.5 | 6% |
| Calibrator3 | 180 | 1077.3 | 17.1 | 2% | 14.9 | 1% | 47.0 | 4% | 51.8 | 4.8% | 73.5 | 7% |
| Control 1 | 180 | 179.4 | 3.2 | 2% | 1.9 | 1% | 6.1 | 3% | 7.1 | 4.0% | 10.0 | 6% |
| Control 2 | 180 | 609.6 | 13.2 | 2% | 12.6 | 2% | 16.4 | 3% | 24.4 | 4.0% | 34.6 | 6% |

b. Linearity/assay reportable range:

A linearity study was conducted following CLSI EP6-A for Diazyme Fibrinogen Assay on the Modular P analyzer. A human serum sample was spiked with fibrinogen stock solution to a final concentration of $1032.8\mathrm{mg / dL}$ Fibrinogen. The final concentration of the high sample pool was determined using Diazyme Fibrinogen assay with triplicate analysis. The prepared high sample pool was diluted with saline to create the linearity samples.

The linearity samples were tested with Diazyme Fibrinogen Assay in triplicate and the results are provided below.

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| Sample Recovery Results | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Level | Recovery Triplicate mg/dL | | | Recover Mean mg/dL | Expected Value mg/dL | Error mg/dL | % Recovery |
| 0 | 0 | 0.7 | 0.0 | 0.4 | 0.0 | -0.37 | NMA |
| 1 | 108.1 | 110.1 | 103.3 | 109.1 | 103.3 | -5.82 | 105.63% |
| 2 | 198.7 | 197.9 | 206.6 | 197.1 | 206.6 | 9.43 | 95.43% |
| 3 | 301.1 | 296.2 | 309.9 | 296.3 | 309.9 | 13.55 | 95.63% |
| 4 | 380.1 | 385.8 | 413.1 | 385.9 | 413.1 | 27.27 | 93.40% |
| 5 | 533.7 | 521.3 | 516.4 | 527.1 | 516.4 | -10.68 | 102.07% |
| 6 | 622.1 | 622.9 | 619.7 | 620.7 | 619.7 | -1.00 | 100.16% |
| 7 | 740.1 | 716.6 | 723.0 | 728.0 | 723.0 | -5.02 | 100.69% |
| 8 | 859 | 844.6 | 826.3 | 847.0 | 826.3 | -20.70 | 102.51% |
| 9 | 940.1 | 954.4 | 929.6 | 952.9 | 929.6 | -23.35 | 102.51% |
| 10 | 1025.6 | 1035.4 | 1032.8 | 1032.8 | 1032.8 | 0.00 | 100.00% |

The results showed that the best fitting model for Diazyme Fibrinogen Assay is linear up to $1032.8\mathrm{mg / dL}$ and the recovery mean values for each level were within $10\%$ of the expected values. The AMR of Diazyme Fibrinogen assay is claimed to be $100 - 900\mathrm{mg / dL}$ .

c. Traceability, Stability, Expected values (controls, calibrators, or methods):

# Traceability:

Master Calibrator (reference calibrator) Preparation and Value Assignment

i. The reference calibrators were prepared by spiking fibrinogen to diluted human serum to target concentrations. The value point of the three levels of the calibrators is set across the reportable range. The highest calibrator is set to close to the upper measuring range. Since the calibration curve is nonlinear, manufacturing targets for each level is designed for best curve fit.

The initial value of the fibrinogen at each level was assayed by the predicate method, Kamiya Biomedical K-Assay Fibrinogen. The initial assigned value of the master calibrators was finalized after verification testing with WHO fibrinogen reference materials NIBSC code 09/242. The WHO fibrinogen reference material was used to prepare the stock solution (1500 mg/dL) and three diluted fibrinogen solutions (375, 500, and 750 mg/dL). After calibration with the Diazyme Fibrinogen Calibrator Set, the reference material dilutions were tested with the Diazyme Fibrinogen Assay in duplicate on the Modular P analyzer. The result showed that the WHO reference material dilutions recovered within ±10% of the target values. Therefore, Diazyme Fibrinogen Calibrators of the Diazyme Fibrinogen Assay are traceable to WHO reference material NIBSC code 09/242.

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| Master Calibrator Value Assignment Data | | | | | | |
| --- | --- | --- | --- | --- | --- | --- |
| Sample | Expected mg/dL | Dilution Factor | Replicate 1 mg/dL | Replicate 2 mg/dL | Mean mg/dL | % Recovery |
| 09/242 | 375 | 4 | 365.0 | 362.1 | 363.6 | 96.9% |
| 09/242 | 500 | 3 | 524.1 | 512.4 | 518.3 | 103.7% |
| 09/242 | 750 | 2 | 773.5 | 771.9 | 772.7 | 103.0% |
| 09/242 | 1500 | No dilution | | | | |

## Production Calibrator Preparation and Value Assignment

ii. Production calibrator and value assignment is prepared from a calculated amount of human fibrinogen stock solution is added into a diluted serum base to target concentrations. The value point of the three levels of the calibrators is set across the reportable range. The highest calibrator is set close to the upper measuring range. Since the calibration curve is non-linear, manufacturing targets for each level is designed for best curve fit. The production calibrators are value assigned by use of master calibrator and Diazyme reagent with multiple testing runs in three days.

The mean value of each level was used to calibrate Diazyme Fibrinogen Assay.

## Production Control Preparation and Value Assignment

Production controls are prepared by spiking fibrinogen stock solution to a diluted human serum to the target fibrinogen concentrations. For the level 1 (normal) control the manufacturing target is approximately 180 mg/dL. For level 2 (abnormal) control the manufacturing target is approximately 600 mg/dL. The Diazyme Fibrinogen Assay and calibrators are used in replicate analysis to determine the mean value of the newly prepared controls. Testing was performed with one Modular P analyzer in two runs in duplicate for three days. The final value is the mean of replicate values and the expected range is calculated as ±15% from the mean value for the respective controls.

## Stability:

### Reagent Shelf-life Stability:

Three lots of Diazyme Fibrinogen Assay reagents were used to determine shelf-life stability. The reagents from each lot were kept in incubators at 37°C and 25°C for the accelerated stability study and 2-8°C for the real-time stability study. At the indicated time, the Diazyme Fibrinogen Assay reagents were removed from the incubators and evaluated by testing six citrated human plasma samples containing fibrinogen covering the claimed AMR after each calibration. The acceptance criterion was sample value recovery within 10% deviation from the results at day 0.

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| Reagent Accelerated Stability (37°C/3 lots) | | | | | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Day | Sample 1 | | Sample 2 | | Sample 3 | | Sample 4 | | Sample 5 | | Sample 6 | |
| | Res | Rec | Res | Rec | Res | Rec | Res | Rec | Res | Rec | Res | Rec |
| 0 | 452. | 99.7 | 327.9 | 98.4 | 206.3 | 98.9 | 105.9 | 98.7 | 703.8 | 99.6 | 823.3 | 99.9 |
| 3 | 456. | 100.6 | 323.6 | 97.1 | 200.3 | 96.0 | 102.9 | 95.9 | 731.2 | 103.5 | 869.5 | 105.5 |
| 7 | 442. | 97.5 | 323.3 | 97.0 | 203.4 | 97.5 | 103.5 | 96.5 | 683.5 | 96.7 | 796.8 | 96.7 |
| 11 | 436. | 96.4 | 321.5 | 96.5 | 205.8 | 98.6 | 105.3 | 98.1 | 670.9 | 94.9 | 798.3 | 96.9 |
| 14 | 442. | 97.5 | 327.2 | 98.2 | 207.2 | 99.3 | 106.7 | 99.4 | 676.2 | 95.7 | 801.1 | 97.2 |
| 15 | 426. | 94.1 | 315.6 | 94.7 | 200.9 | 96.2 | 102.1 | 95.1 | 684.1 | 96.8 | 815.3 | 99.0 |

Res: Result (mg/dL)
Rec: Recovery $(\%)$

| Reagent Accelerated Stability (25°C/3 lots) | | | | | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Months | Sample 1 | | Sample 2 | | Sample 3 | | Sample 4 | | Sample 5 | | Sample 6 | |
| | Res | Rec | Res | Rec | Res | Rec | Res | Rec | Res | Rec | Res | Rec |
| 0 | 426. | 94.1 | 319.0 | 95.7 | 197.8 | 94.8 | 103.5 | 96.5 | 709.3 | 100.4 | 831.7 | 100.9 |
| 1 | 445. | 98.2 | 329.4 | 98.8 | 208.1 | 99.7 | 107.1 | 99.8 | 686.0 | 97.1 | 795.7 | 96.6 |
| 3 | 415. | 91.6 | 303.8 | 91.1 | 192.1 | 92.0 | 98.4 | 91.7 | 666.7 | 94.3 | 782.3 | 94.9 |

Res: Result (mg/dL)
Rec: Recovery $(\%)$

The results demonstrated that the reagents were stable for 14 days when stored at $37^{\circ}\mathrm{C}$ and stable for over 3 months when stored at $25^{\circ}\mathrm{C}$ . According to an Arrhenius equation the reagent shelf-life is calculated to 18 months when kept at $2 - 8^{\circ}\mathrm{C}$ . The real time stability at $2 - 8^{\circ}\mathrm{C}$ is on-going and final results will be reported.

# Reagent On-Board Stability on Roche Modular P:

Three lots of reagents were evaluated for on-board stability on the Roche Modular P analyzer. The reagents were kept in the Roche Modular reagent bottle in the $2 - 8^{\circ}\mathrm{C}$ chamber of the instrument. In this study, six citrated human plasma samples were tested after each calibration on the indicated days. The corresponding testing days and results are listed in the table below. The acceptance criterion was sample value recovery within $10\%$ deviation from day 0.

| Reagent On-Board Stability (2-8°C/3 lots) | | | | | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Day | Sample 1 | | Sample 2 | | Sample 3 | | Sample 4 | | Sample 5 | | Sample 6 | |
| | Res | Rec | Res | Rec | Res | Rec | Res | Rec | Res | Rec | Res | Rec |
| 0 | 426. | 94.1 | 319.0 | 95.7 | 197.8 | 94.8 | 103.5 | 96.5 | 709.3 | 100.4 | 831.7 | 100.9 |
| 5 | 452. | 99.7 | 327.1 | 98.1 | 200.3 | 96.0 | 101.8 | 94.9 | 755.6 | 106.9 | 872.6 | 105.9 |
| 10 | 440. | 97.2 | 327.0 | 98.1 | 208.4 | 99.8 | 106.6 | 99.3 | 676.5 | 95.7 | 803.2 | 97.5 |
| 15 | 448. | 98.9 | 325.5 | 97.6 | 205.9 | 98.7 | 105.2 | 98.0 | 688.8 | 97.5 | 821.4 | 99.7 |
| 21 | 445. | 98.2 | 329.4 | 98.8 | 208.1 | 99.7 | 107.1 | 99.8 | 686.0 | 97.1 | 795.7 | 96.6 |
| 24 | 436. | 96.2 | 323.6 | 97.1 | 204.8 | 98.1 | 104.7 | 97.5 | 677.6 | 95.9 | 796.0 | 96.6 |
| 25 | 439. | 96.9 | 319.2 | 95.8 | 201.6 | 96.6 | 102.1 | 95.1 | 688.6 | 97.4 | 799.3 | 97.0 |

Res: Result (mg/dL)
Rec: Recovery $(\%)$

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Based on the results above, the reagent is stable for 20 days on board the Roche Modular P analyzer.

## Calibrator and Control Stability Test:

Accelerated Stability at 37°C

Three lots of the Diazyme Fibrinogen Calibrators and two lots of the Controls were kept in a 37°C incubator. At the indicated time, the calibrators and Controls were removed, reconstituted, and tested as samples with Diazyme Fibrinogen Assay reagent and master calibrators stored at 2-8°C on the Roche Modular P analyzer. The acceptance criterion was calibrator and control value recovery within 10% deviation from day 0.

| Calibrator and Controls Accelerated Stability Study (37°C, 3 lots) | | | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Day | Calibrator | | | | | | Control | | | |
| | Level 1 | | Level 2 | | Level 3 | | Level 1 | | Level 2 | |
| | Res | Rec | Res | Rec | Res | Rec | Res | Rec | Res | Rec |
| 0 | 253. | 100.0 | 556.9 | 100.0 | 1098. | 100.0 | 176.2 | 100.0 | 675.6 | 100.0 |
| 3 | 259. | 102.7 | 562.1 | 100.9 | 1177. | 107.1 | 184.0 | 104.4 | 686.6 | 101.6 |
| 7 | 260. | 103.0 | 531.9 | 95.5 | 1062. | 96.6 | 186.1 | 105.7 | 650.5 | 96.3 |
| 11 | 260. | 103.3 | 528.7 | 94.9 | 1101. | 100.2 | 186.4 | 105.8 | 669.2 | 99.1 |
| 14 | 262. | 103.6 | 531.5 | 95.4 | 1121. | 102.0 | 189.3 | 107.5 | 645.6 | 95.6 |
| 15 | 263. | 104.2 | 533.6 | 95.8 | 1155. | 105.2 | 184.1 | 104.5 | 658.7 | 97.5 |

Res: Result (mg/dL)
Rec: Recovery (%)

Results demonstrated that the calibrators and controls were stable for 14 days when stored at 37°C. According to an Arrhenius equation the reagent shelf-life is calculated to 18 months when kept at 2-8°C.

## Real Time Stability (2-8°C)

The real time stability studies at 2-8°C for calibrators and controls are on-going and the final results will be reported. The acceptance criterion is calibrator and control value recovery within 10% deviation from day 0.

## Reconstitution Stability Assay at 2-8°C

Three lots of the Diazyme Fibrinogen Calibrators and two lots of Controls were reconstituted and kept in a 2-8°C incubator. At the indicated time, the calibrators and Controls were removed and tested as samples with Diazyme Fibrinogen reagent (one lot) and master calibrators stored at 2-8°C.

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| Calibrator and Control Reconstitution Stability Study (2-8°C, 3 lots) | | | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Day | Calibrator | | | | | | Control | | | |
| | Level 1 | | Level 2 | | Level 3 | | Level 1 | | Level 2 | |
| | Res | Rec | Res | Rec | Res | Rec | Res | Rec | Res | Rec |
| 0 | 255. | 100.0 | 556.9 | 100.0 | 1115. | 100.0 | 182.8 | 100.0 | 675.5 | 100.0 |
| 3 | 263. | 103.0 | 570.9 | 102.5 | 1185. | 106.3 | 188.5 | 103.1 | 680.8 | 100.8 |
| 5 | 266. | 104.3 | 550.8 | 98.9 | 1159. | 104.0 | 188.8 | 10.32 | 675.2 | 100.0 |
| 7 | 259. | 101.6 | 524.0 | 94.1 | 1015. | 91.0 | 185.7 | 101.5 | 625.9 | 92.6 |
| 8 | 251. | 98.4 | 526.7 | 94.6 | 1008. | 90.4 | 187.2 | 102.4 | 622.9 | 92.2 |

Res: Result (mg/dL)
Rec: Recovery (%)

The results listed in the table above demonstrated that the calibrators and controls were stable for 7 days after reconstitution and stored at 2-8°C.

## Sample stability (2-8°C real-time stability)

Six citrated plasma samples covering the whole AMR were stored at 2-8°C for one and two days and used for a real-time patient sample stability study. At the indicated time points, samples were tested with Diazyme Fibrinogen Assay on the Roche Modular P analyzer. The results are listed in the table below.

| Patient Sample Real-time Stability Data | | | | | | |
| --- | --- | --- | --- | --- | --- | --- |
| Sample ID | 0 Days | | 1 Days | | 2 Days | |
| | Results mg/dL | Recovery % | Results mg/dL | Recovery % | Results mg/dL | Recovery % |
| Plasma 1 | 420.0 | 100% | 428.6 | 102% | 453.40 | 108% |
| Plasma 2 | 294.8 | 100% | 311.0 | 105% | 319.60 | 108% |
| Plasma 3 | 204.7 | 100% | 205.2 | 100% | 206.6 | 101% |
| Plasma 4 | 103.5 | 100% | 107.7 | 104% | 107.4 | 104% |
| Plasma 5 | 722.3 | 100% | 699.5 | 97% | 717.00 | 99% |
| Plasma 6 | 826.3 | 100% | 818.6 | 99% | 846.50 | 102% |

After storage of 1 day and 2 days, all six samples recovered within 10% deviation from day 0. The pre-determined acceptance criterion was met.

## Patient Sample Freeze-Thaw Stability

Six fresh citrated plasma samples covering the whole AMR were tested before and after one and two freeze-thaw cycles with one lot Diazyme Fibrinogen Assay reagent on the Roche Molecular P analyzer. The acceptance criterion was the recovery after freeze within 10% deviation from before freezing. Results demonstrated that all six patient samples were not affected by one freeze-thaw cycle.

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| Patient Sample Freeze-Thaw Stability Data | | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| Sample ID | Before Freeze mg/dL | Recovery | Freeze/Thaw Cycle 1 mg/dL | Recovery | Pass/Fail | Freeze/Thaw Cycle 2 mg/dL | Recovery | Pass/Fail |
| Sample 1 | 444.5 | 100% | 445.8 | 100% | Pass | 471.9 | 106% | Pass |
| Sample 2 | 321.7 | 100% | 316.1 | 98% | Pass | 330.6 | 103% | Pass |
| Sample 3 | 199.0 | 100% | 203.0 | 102% | Pass | 211.1 | 106% | Pass |
| Sample 4 | 102.9 | 100% | 102.8 | 100% | Pass | 105.2 | 102% | Pass |
| Sample 5 | 683.7 | 100% | 689.4 | 101% | Pass | 728.8 | 107% | Pass |
| Sample 6 | 819.8 | 100% | 802.1 | 98% | Pass | 826.4 | 101% | Pass |

d. Detection limit:

The Limit of Blank (LoB) and Limit of Detection (LoD) of the Diazyme Fibrinogen Assay were determined by testing on Roche Modular P analyzer with three lots of reagents and three lots of calibrators according to CLSI EP17-A2. True blank is 7.5% bovine serum albumin (BSA) solution in saline with 60 replicates for LoB determination. To determine LoD, five low level positive plasma samples were diluted with 7.5% BSA in 0.9% saline to a targeted low sample range using 12 replicates.

$$
\mathrm{LoD} = \mathrm{LoB} + (1.645 \times \mathrm{SD} \text{ of LoD low sample})
$$

The LoQ of the Diazyme Fibrinogen Assay was determined by following CLSI EP17-A2. Five citrated plasma samples were diluted with 7.5% BSA to concentrations of 1 to 20 times of assumed LoB. The diluted citrated plasma samples were tested with three lots of the Diazyme Fibrinogen Assay reagents on the Roche Modular P analyzer. The results are summarized below.

| LoB, LoD and LoQ (3 lots reagents) | | |
| --- | --- | --- |
| LoB | LoD | LoQ |
| 2.15 mg/dL | 5.08 mg/dL | 12.9 mg/dL |

e. Analytical specificity:

i. To determine the level of interference from substances present in citrated plasma, the Diazyme Fibrinogen Assay was used to test three citrated plasma samples, which contain low (100 mg/dL), medium (300 mg/dL), and high (800 mg/dL) fibrinogen concentrations spiked with various concentrations of substances according to CLSI EP7-A2 guideline. Each spiked sample was tested in triplicate on the Roche Modular P analyzer using one lot of Diazyme Fibrinogen Assay reagent and one lot of calibrator. The acceptance criterion was the difference of mean fibrinogen concentration between the spiked sample and the un-spiked sample < ± 10%.

The common endogenous and exogenous interfering substances and their interference results are listed below and showed no significant interference (< ± 10%) up to the indicated concentration for all three tested citrated plasma samples at 100 mg/dL, 300 mg/dL and 800 mg/dL, respectively. The

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additional interference studies for anticoagulant medications were also performed.

| Endogenous Substances | |
| --- | --- |
| Interferent | Concentration |
| Ascorbic Acid | 176 mg/dL |
| Bilirubin | 40 mg/dL |
| Bilirubin Conjugated | 40 mg/dL |
| Hemoglobin | 1000 mg/dL |
| Rheumatoid Factor | 220 IU/ml |
| Triglycerides | 1000 mg/dL |
| Exogenous Substances | |
| --- | --- |
| Interferent | Concentration |
| Acetylsalicylic Acid | 2.78 mM |
| Na2-Cefoxitin | 1554 μM |
| Ibuprofen | 2438 μM |
| Unfractionated Heparin | 3000 U/L |
| Low Molecular Weight Heparin | 3000 U/L |
| Additional Interference | |
| --- | --- |
| Interference | Concentration |
| Warfarin | 65 μM |
| Dabigatran | 3.7 μg/mL |
| Hirudin | 25 μg/mL |
| Rivaroxaban | 7.0 μg/mL |
| Fibrin Degradation Product | 0.5 mg/mL |
| Argatroban | 20 μg/mL |

ii. High Dose Hook Effect

To test for High Dose "Hook Effect" associated with the fibrinogen assay, an 11-level linearity set was prepared with the highest level containing 1426.0 mg/dL fibrinogen. The linearity set prepared was first tested with Diazyme Fibrinogen Assay on the Roche Modular P in triplicate with the normal dilution ratio of 1:20. The value recovery data is shown below.

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| Hook Effect, 1:20 Dilution | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Level | rep 1 mg/dL | rep 2 mg/dL | rep 3 mg/dL | Average mg/dL | Expected mg/dL | Error mg/dL | % Error |
| 1 | 25.6 | 22.8 | 21.8 | 23.4 | 22.8 | 0.58 | -2.50% |
| 2 | 52.5 | 49.4 | 51.5 | 51.1 | 47.5 | 3.60 | -7.04% |
| 3 | 98.1 | 102.6 | 99.9 | 100.2 | 95.1 | 5.14 | -5.13% |
| 4 | 187.6 | 190.6 | 191.5 | 189.9 | 190.1 | -0.23 | 0.12% |
| 5 | 269.8 | 289.7 | 278.9 | 279.5 | 285.2 | -5.72 | 2.05% |
| 6 | 365.2 | 356.0 | 354.4 | 358.5 | 380.3 | -21.72 | 6.06% |
| 7 | 471.3 | 451.7 | 451.0 | 458.0 | 475.3 | -17.32 | 3.78% |
| 8 | 700.1 | 695.2 | 696.1 | 697.1 | 713.0 | -15.84 | 2.27% |
| 9 | 929.5 | 932.6 | 937.4 | 933.2 | 950.6 | -17.47 | 1.87% |
| 10 | 1152.3 | 1171.0 | 1156.7 | 1160.0 | 1188.3 | -28.30 | 2.44% |
| 11 | 1393.2 | 1320.7 | 1376.7 | 1363.5 | 1426.0 | -62.42 | 4.58% |

Due to the solubility limit of fibrinogen antigen, extremely high concentrations of the analyte could not be stably prepared. The same linearity set prepared above was tested again with a 1:5 sample dilution. The test results showed that the sample containing theoretical fibrinogen concentration of 4990 mg/dL was recovered as 1715.1 mg/dL. This value was still greater than the upper end of the AMR of 900 mg/dL. Therefore, the Diazyme Fibrinogen Assay demonstrated no hook effect at least at 4990 mg/dL Fibrinogen concentration.

| Hook Effect, 1:5 Dilution | | | | | | | |
| --- | --- | --- | --- | --- | --- | --- | --- |
| Level | rep 1 mg/dL | rep 2 mg/dL | rep 3 mg/dL | Average mg/dL | Expected mg/dL | Error mg/dL | % Error |
| 12 | 84.5 | 88.1 | 87.9 | 86.8 | 79.9 | 7.0 | -8.04% |
| 13 | 196.8 | 182.4 | 170.4 | 183.2 | 166.4 | 16.8 | -9.19% |
| 14 | 315.8 | 328.9 | 324.4 | 323.0 | 332.7 | -9.7 | 3.00% |
| 15 | 671.0 | 666.2 | 687.2 | 674.8 | 665.5 | 9.4 | -1.39% |
| 16 | 979.9 | 1062.9 | 1046.4 | 1029.7 | 998.2 | 31.6 | -3.07% |
| 17 | 1195.1 | 1254.4 | 1339.9 | 1263.1 | 1330.9 | -67.8 | 5.36% |
| 18 | 1525.9 | 1453.5 | 1442.0 | 1473.8 | 1663.6 | -189.8 | 12.88% |
| 19 | 1789.9 | 1791.5 | 1826.6 | 1802.7 | 2495.4 | -692.8 | 38.43% |
| 20 | 1855.5 | 1866.7 | 1884.8 | 1869.0 | 3327.2 | -1458.2 | 78.02% |
| 21 | 1835.9 | 1856.8 | 1827.2 | 1840.0 | 4159.0 | -2319.1 | 126.04% |
| 22 | 1718.0 | 1707.0 | 1720.2 | 1715.1 | 4990.8 | -3275.8 | 191.00% |

f. Assay cut-off:

Not applicable

2. Comparison studies:

a. Method comparison with predicate device:

To demonstrate accuracy, one lot of Diazyme Fibrinogen Assay reagent was tested on the Roche Modular P analyzer with one lot of the Diazyme Fibrinogen calibrator on

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individual citrated plasma samples from the intended target population. One lot of Kamiya K-Assay Fibrinogen reagent was used as the predicate and tested on the Hitachi 917 on the same citrated plasma samples. CLSI EP9-A3 was used as a guideline for the method comparison studies. All samples were tested in singlet with both methods. Sample demographic information was included as provided by the collection center. All samples were collected in $3.2\%$ citrate tubes.

A total of 176 unique samples were tested across the three sites and the acceptance criteria were: Slope $= 1.0 \pm 0.1$ ; $r^2 \geq 0.95$ ; y-intercept $< \mathrm{LoQ}$ . The comparison results obtained by both methods are tabulated below:

| Regression Analysis (Regular Linear) | | | | |
| --- | --- | --- | --- | --- |
| Parameter | Site 1 | Site 2 | Site 3 | Total |
| N | 59 | 59 | 58 | 176 |
| Slope | 0.972 | 0.983 | 1.004 | 0.986 |
| 95% CI | 0.953 – 0.992 | 0.965 – 1.000 | 0.984 – 1.023 | 0.975 – 0.997 |
| Intercept | 7.20 | 1.54 | -0.41 | 2.72 |
| 95% CI | -1.03 – 15.43 | -5.40– 8.48 | -8.18 – 7.37 | -1.70 – 7.13 |
| Standard Error of Estimate | 15.69 | 13.74 | 15.75 | 15.31 |
| Correlation Coefficient(R) | 0.9972 | 0.9978 | 0.9973 | 0.9973 |
| R² | 0.9944 | 0.9956 | 0.9946 | 0.9946 |
| Sample Range (Diazyme) | 119.6 – 825.6 | 107.8 – 869.9 | 107.5 – 870.0 | 107.5 – 870.0 |
| Regression Analysis (Deming Regression) | | | | |
| --- | --- | --- | --- | --- |
| Parameter | Site 1 | Site 2 | Site 3 | Total |
| n | 59 | 59 | 58 | 176 |
| Slope | 0.975 | 0.985 | 1.006 | 0.989 |
| 95% CI | 0.956 – 0.994 | 0.968 – 1.002 | 0.987 – 1.026 | 0.978 – 0.999 |
| Intercept | 6.22 | 0.81 | -1.31 | 1.80 |
| 95% CI | -2.01– 14.45 | -6.14 – 7.75 | -9.09 – 6.48 | -2.68 – 6.22 |
| Standard Error of Estimate | 15.70 | 13.75 | 15.76 | 15.32 |
| Correlation Coefficient(R) | 0.9972 | 0.9978 | 0.9973 | 0.9973 |
| R² | 0.9944 | 0.9956 | 0.9946 | 0.9946 |
| Sample Range (Diazyme) | 119.6 – 825.6 | 107.8 – 869.9 | 107.5 – 870.0 | 107.5 – 870.0 |

The method comparison study results met acceptance criteria.

# b. Matrix comparison:

Not applicable

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3. Clinical studies:
a. Clinical Sensitivity:
Not applicable
b. Clinical specificity:
Not applicable
c. Other clinical supportive data (when a. and b. are not applicable):
Not applicable

4. Clinical cut-off:
Not applicable

5. Expected values/Reference range:
To establish the reference interval for the Diazyme Fibrinogen Assay, citrated plasma samples from 120 apparently healthy adults were tested in singlet using the Diazyme Fibrinogen Assay on the Roche Modular P analyzer according to CLSI C28-A3 guideline. The established reference interval is from 130.7 to 397.5 mg/dL.

N. Proposed Labeling:
The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

O. Conclusion:
The submitted information in this premarket notification is completed and supports a substantial equivalence decision.

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