Verification of an immunoturbidimetric assay for heart-type fatty acid-binding protein (H-FABP) on a clinical chemistry platform and establishment of the upper reference limit

Background: Heart-type fatty acid-binding protein (H-FABP) is an early biomarker of cardiac injury. Randox Laboratories developed an immunoturbidimetric H-FABP assay for non-proprietary automated clinical chemistry analysers that could be useful in the emergency department. We verified the analytical performances claimed by Randox Laboratories on Roche Cobas 6000 clinical chemistry platform in use in our laboratory, and we defined our own 99th percentile upper reference limit for H-FABP. Methods: For the verification of method performances, we used pools of spared patient samples from routine and two levels of quality control material, while samples for the reference value study were collected from 545 blood donors. Following CLSI guidelines we verified limit of blank (LOB), limit of detection (LOD), limit of quantitation (LOQ), repeatability and within-laboratory precision, trueness, linearity, and the stability of H-FABP in EDTA over 24. h. Results and discussion: The LOQ (3.19. μg/L) was verified with a CV% of 10.4. The precision was verified for the low (mean 5.88. μg/L, CV. =. 6.7%), the medium (mean 45.28. μg/L, CV. =. 3.0%), and the high concentration (mean 88.81. μg/L, CV. =. 4.0%). The trueness was verified as well as the linearity over the indicated measurement interval of 0.747-120. μg/L. The H-FABP in EDTA samples is stable throughout 24. h both at room temperature and at 4. °C. The H-FABP 99th percentile upper reference limit for all subjects (3.60. μg/L, 95% CI 3.51-3.77) is more appropriate than gender-specific ones that are not statistically different.