Rapid genomic typing of BK virus directly from clinical specimens.

Abstract:

: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.

journal_name

Mol Cell Probes

authors

Jin L

doi

10.1006/mcpr.1993.1047

subject

Has Abstract

pub_date

1993-08-01 00:00:00

pages

331-4

issue

4

eissn

0890-8508

issn

1096-1194

pii

S0890-8508(83)71047-9

journal_volume

7

pub_type

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