Analytical Validation of Familial Hypercholesterolemia Biomarkers in Dried Blood Spots

International Journal of Neonatal Screening

Michael Lasarev¹  Vanessa Horner²  Amy Peterson³  Patrice K. Held³  Kristin Campbell⁴  Amy E. Wiberley-Bradford⁴ 

[1] Department of Biostatistics and Medical Informatics, School of Medicine and Public Health, University of Wisconsin, Madison, WI 53706, USA;Department of Pathology, School of Medicine and Public Health, University of Wisconsin, Madison, WI 53706, USA;Department of Pediatrics, School of Medicine and Public Health, University of Wisconsin, Madison, WI 53706, USA;Wisconsin State Laboratory of Hygiene, School of Medicine and Public Health, University of Wisconsin, Madison, WI 53706, USA;

关键词: newborn screening;    familial hypercholesterolemia;    cholesterol;    low-density lipoprotein;    apolipoprotein B;    dried blood spots;   

DOI  :  10.3390/ijns8010014

来源: DOAJ

【 摘 要 】

Heterozygous familial hypercholesterolemia (HeFH) is a common, treatable genetic disorder characterized by premature atherosclerosis and cardiovascular disease, yet the majority of affected individuals remain undiagnosed. Newborn screening could play a role in identification of at-risk individuals and provide an opportunity for early intervention, prior to the onset of symptoms. The objective of this study was to develop and validate assays for quantification of candidate HeFH biomarkers in dried blood spots (DBS). Commercially available enzyme assay kits for quantification of serum total cholesterol (TC) and low-density lipoprotein-cholesterol (LDL-C) were modified for high-throughput analysis of DBS. Apolipoprotein B (ApoB) concentrations in DBS were measured using an immunoassay with modifications from published studies. All three assays were validated according to the College of American Pathologists guidelines for clinical laboratories. The performance of TC, LDL-C, and ApoB assays was assessed by precision, recovery, limit of quantification (LOQ) and linearity. Precision studies yielded coefficients of variation (CV) of less than 15%, with recovery greater than 75% for all three assays. The determined LOQ and linearity were comparable to serum-based assays. In a direct comparison between serum and DBS concentrations, positive correlations were demonstrated for TC, LDL-C, and ApoB. Additionally, the initial evaluation of the three biomarker concentrations within the unaffected population was similar to values obtained in previous published studies. This study reports on methods for quantification of TC, LDL-C, and ApoB in DBS. Assay validation results were within acceptable limits for newborn screening. This is an important first step toward the identification of newborns with HeFH.

【 授权许可】

Unknown