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A rapid and efficient method BK polyomavirus genotyping by high‐resolution melting analysis

A small percentage of renal patients become infected with the BK polyomavirus (BKV), a pathogenic virus that causes BKV‐associated nephropathy (BKVN), after kidney transplantation. This study presents a simple, rapid, high‐throughput method for BKV genotyping using high‐resolution melt analysis (HRMA). Using this novel method, BKV genotypes were analyzed in 49 samples taken from BKV‐positive renal transplantation patients for classification into 1 of 3 genotypes: GI‐1 (subgroups Ia, Ib1, and Ic), GI‐2 (subgroup Ib2), and GII‐IV (subtypes II, III, and IV). HRMA was performed to compare each sample sequence to a reference sequence that contained a combination of 2 of the 3 genotype groups, and the findings validated by conventional DNA sequencing. Of the 49 samples, 20 samples were classified as GI‐1, 18 as GI‐2, and 11 as GII‐IV, suggesting that the predominant BKV strain (77.6%) in these patients was subtype I (GI‐1 and GI‐2). The HRMA method presented here is a time‐saving, reliable, and low‐cost procedure that can be developed as a diagnostic tool in the detection of the specific BKV genotypes associated with BKVN. J. Med. Virol. 83:2128–2134, 2011. © 2011 Wiley Periodicals, Inc.