257. A Rapid Genotyping Method for Polyomavirus BK
Session: Poster Abstract Session: Diagnostic Microbiology; Novel Molecular Methods
Thursday, October 3, 2013
Room: The Moscone Center: Poster Hall C
Posters
  • ID week 40364 A Rapid Genotyping Method for Polyomavirus BK.pdf (101.8 kB)
  • Background:

    Polyomavirus BK (BKV) is ubiquitous and widely spread in adult human populations. Infection results in lifelong persistence of the virus. BKV may cause nephropathy in renal transplant recipients and hemorrhagic cystitis in bone marrow recipients. We developed an easy to use genotyping tool to investigate the relation of BKV genotype (I-IV) with nephropathy and hemorrhagic cystitis.

    Methods:

    A multiplex of Real Time PCRs (RT-PCR) was developed and validated on the VP1 gene to differentiate the 4 main genotypes of BKV. 150 BKV positive samples (17 plasma, 133 urine) were tested with these specific assays. Of these 150 samples, 50 were additionally confirmed by sequencing the 1630-1956 nucleotide fragment.

    Results:

    For every genotype, a 100% specific and internally controlled assay was developed. The precision of the 4 RT-PCRs remained within 1SD and the limit of detection was log 3 copies/ml. Of the 150 BKV positive samples, 105 (70%) were genotype I, 8 (5.3%) genotype II, 19 (12.7%) genotype IV and 3 (2%) genotype I+IV. In 15 (10%) samples genotyping was not successful due to a low viral load. By sequence analysis the genotypes of 46 of the 50 and 2 of the 3 samples with the double infection could be confirmed.

    Conclusion: This study describes a new multiplex RT-PCR for detection of the subtypes of BKV. It proved to be a rapid, cheap and sensitive genotyping tool compared to sequencing, and can detect double infections with different BK genotypes. This method will be of value to obtain insight in the relation of BKV genotype with nephropathy and hemorrhagic cystitis.

    Lilli Gard, BSc, Bert Niesters, PhD and Annelies Riezebos-Brilman, MD, PhD, Medical Microbiology, University Medical Center Groningen, Groningen, Netherlands

    Disclosures:

    L. Gard, None

    B. Niesters, None

    A. Riezebos-Brilman, None

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