Rapid genomic typing of BK virus directly from clinical specimens.
A simple and rapid method, PCR-restriction enzyme analysis (PCR-RE) for BK virus (BKV) typing was developed, based on the presence of type-specific restriction enzyme sites in a 327 bp PCR-generated fragment which partially encodes the VP1 protein. The enzymes, Alu I, Xmn I and Ava II, were used to digest the PCR products in two stages. Ethidium bromide banding patterns characteristic of each of four subtypes of BKV were visualized through gel electrophoresis. A total of 37 samples from clinical specimens and culture fluids were successfully subtyped using the PCR-RE. A second method, PCR-sequencing, was applied to samples that generated less than 100 ng of DNA. These were subjected to a second round of PCR and then sequenced from single stranded templates immobilised via a biotinylated primer. The subtypes were assigned on the basis of the type-specific sequences previously characterized.
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Virus Reference Division, Central Public Health Laboratory, London, UK.
D001739:BK Virus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D001483:Base Sequence; D005260:Female; D016679:Genome, Viral; D006801:Humans; D007165:Immunosuppression Therapy; D008969:Molecular Sequence Data; D030361:Papillomavirus Infections / Q000150:complications / Q000235:genetics / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D011251:Pregnancy Complications, Infectious; D015183:Restriction Mapping; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid; D014412:Tumor Virus Infections / Q000150:complications / Q000235:genetics / Q000382:microbiology