Analytical validation and performance characteristics of molecular serum biomarkers, miR-371a-3p and miR-372-3p, for male germ cell tumors, in a clinical laboratory setting Journal Article

   

Authors:	Ye, F.; Feldman, D. R.; Valentino, A.; So, R.; Bromberg, M.; Khan, S.; Funt, S. A.; Sheinfeld, J.; Solit, D. B.; Pessin, M. S.; Peerschke, E. I.
Article Title:	Analytical validation and performance characteristics of molecular serum biomarkers, miR-371a-3p and miR-372-3p, for male germ cell tumors, in a clinical laboratory setting
Abstract:	Detection of serum embryonic miRNAs miR-371a-3p and miR-372-3p has been proposed to aid in diagnosis, prognosis, and management of patients with testicular germ cell tumors (GCTs). This study describes the analytical validation and performance of a laboratory-developed test to detect these miRNA targets by stem loop real-time quantitative RT-PCR (RT-qPCR) in serum from patients with GCTs. The assay was standardized using an exogenous spike-in control of nonhuman miRNA from Caenorhabditis elegans (cel-miR-39-3p) to assess extraction efficiency, and an endogenous housekeeping miRNA, miR-30b-5p, to control for miRNA normalization. miRNA results were expressed as relative expression level, using the comparative threshold cycle method (2ΔΔCT). Analytical sensitivity of miR-371a-3p and miR-372-3p was 12.5 and 1.25 copies/μL, respectively. Clinical accuracy was evaluated using GCT patients with (n = 34) and without (n = 17) active disease. Positive/negative cutoffs and indeterminate findings were established on the basis of results from healthy volunteers (n = 25) and assay precision. miR-371a-3p and miR-372-3p exhibited a sensitivity of 81.8% and 87.5%, respectively, and a specificity of 100% and 94%, respectively, and an area under the receiver operating characteristic curve of 0.93 and 0.95, respectively. Taken together, RT-qPCR testing for serum miR-371a-3p and miR-372-3p represents a robust, sensitive, and specific clinical assay to aid in the clinical management of patients with GCT. © 2022 Association for Molecular Pathology and American Society for Investigative Pathology
Keywords:	genetics; neoplasm; microrna; tumor marker; testis tumor; testicular neoplasms; neoplasms, germ cell and embryonal; micrornas; humans; human; male; biomarkers, tumor; mirn372 microrna, human; laboratories, clinical
Journal Title:	Journal of Molecular Diagnostics
Volume:	24
Issue:	8
ISSN:	1525-1578
Publisher:	Elsevier Science, Inc.
Date Published:	2022-08-01
Start Page:	867
End Page:	877
Language:	English
DOI:	10.1016/j.jmoldx.2022.04.007
PUBMED:	35934321
PROVIDER:	scopus
PMCID:	PMC9379668
DOI/URL:	https://www.scopus.com/inward/record.uri?eid=2-s2.0-85135549912&doi=10.1016%2fj.jmoldx.2022.04.007&partnerID=40&md5=936ee9e53ff622d8181a8b3eddffda42
Notes:	Article -- Export Date: 1 September 2022 -- Source: Scopus

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