1. Refining the serum miR-371a-3p test for viable germ cell tumor detection.
- Author
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Lafin, John, Scarpini, Cinzia, Amini, Armon, Konneh, Bendu, Howard, Jeffrey, Gerald, Thomas, Nuno, Michelle, Piao, Jin, Savelyeva, Anna, Wang, Zhaohui, Gagan, Jeffrey, Jia, Liwei, Lewis, Cheryl, Murray, Sarah, Sawa, Yun, Margulis, Vitaly, Woldu, Solomon, Strand, Douglas, Coleman, Nicholas, Amatruda, James, Frazier, A, Murray, Matthew, and Bagrodia, Aditya
- Subjects
Humans ,MicroRNAs ,Neoplasms ,Germ Cell and Embryonal ,Teratoma ,Biological Assay ,Hematologic Tests - Abstract
Circulating miR-371a-3p has excellent performance in the detection of viable (non-teratoma) germ cell tumor (GCT) pre-orchiectomy; however, its ability to detect occult disease is understudied. To refine the serum miR-371a-3p assay in the minimal residual disease setting we compared performance of raw (Cq) and normalized (∆Cq, RQ) values from prior assays, and validated interlaboratory concordance by aliquot swapping. Revised assay performance was determined in a cohort of 32 patients suspected of occult retroperitoneal disease. Assay superiority was determined by comparing resulting receiver-operator characteristic (ROC) curves using the Delong method. Pairwise t-tests were used to test for interlaboratory concordance. Performance was comparable when thresholding based on raw Cq vs. normalized values. Interlaboratory concordance of miR-371a-3p was high, but reference genes miR-30b-5p and cel-miR-39-3p were discordant. Introduction of an indeterminate range of Cq 28-35 with a repeat run for any indeterminate improved assay accuracy from 0.84 to 0.92 in a group of patients suspected of occult GCT. We recommend that serum miR-371a-3p test protocols are updated to (a) utilize threshold-based approaches using raw Cq values, (b) continue to include an endogenous (e.g., miR-30b-5p) and exogenous non-human spike-in (e.g., cel-miR-39-3p) microRNA for quality control, and (c) to re-run any sample with an indeterminate result.
- Published
- 2023