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.
Metic Transplantation Laboratory, USC, Keck School of Medicine, Los Angeles, CA, USA. matsuda@den.hokudai.ac.jp
Yasuhiro
Y
Matsuda
Yasir
Y
Qazi
Yuichi
Y
Iwaki
D001739:BK Virus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D005838:Genotype; D006801:Humans; D016030:Kidney Transplantation; D058889:Molecular Typing / Q000379:methods*; D027601:Polyomavirus Infections / Q000821:virology; D017422:Sequence Analysis, DNA; D044366:Transition Temperature; D014180:Transplantation; D014773:Virology / Q000379:methods*
D004279:DNA, Viral