Abstract：
BACKGROUND:Polyomavirus BK (BKV) may cause nephropathy in renal transplant recipients and hemorrhagic cystitis in bone marrow recipients. We developed real-time PCRs (RT-PCR) to determine easily and rapidly the different BKV genotypes (BKGT) (I-IV). METHODS:On the VP1 gene a duplex of RT-PCRs was developed and validated to differentiate the four main BKGT. 212 BKV positive samples (21 plasma, 191 urine) were tested with these specific PCRs. Of these 212 samples, 55 PCR results were additionally confirmed by sequencing a VP1 gene fragment (nucleotide 1630-1956). RESULTS:For every genotype, a highly specific, precise and internally controlled assay was developed with a limit of detection of log 3 copies per ml. In 18 (8.5%) of these samples genotyping was not successful due to a low viral load. By sequence analysis, the genotype of 46 out of 55 and 2 out of 4 samples with double infection could be confirmed. CONCLUSIONS:This study describes RT-PCRs for detection of the main BKGT. It proved to be rapid, cheap and sensitive compared to sequencing. Double infections can also be detected. This method will be of value to investigate the role of BKV infection in relation to the genotype.
journal_name
J Virol Methodsjournal_title
Journal of virological methodsauthors
Gard L,Niesters HG,Riezebos-Brilman Adoi
10.1016/j.jviromet.2015.04.024subject
Has Abstractpub_date
2015-09-01 00:00:00pages
51-6eissn
0166-0934issn
1879-0984pii
S0166-0934(15)00168-8journal_volume
221pub_type
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