Simultaneous detection and differentiation of human Polyomaviruses JC and BK by a rapid and sensitive PCR-ELAHA assay and a survey of the JCV subtypes within an Australian population


Autoria(s): Whiley, David M.; Arden, Katherine E.; Mackay, Ian M.; Syrmis, Melanie W.; Sloots, Theo P.
Contribuinte(s)

Arie J. Zuckerman

Data(s)

01/03/2004

Resumo

Human polyomaviruses JCV and BKV can cause several clinical manifestations in immunocompromised hosts, including progressive multifocal leukoencephalopathy (PML) and haemorrhagic cystitis. Molecular detection by polymerase chain reaction (PCR) is recognised as a sensitive and specific method for detecting human polyomaviruses in clinical samples. In this study, we developed a PCR assay using a single primer pair to amplify a segment of the VP1 gene of JCV and BKV. An enzyme linked amplicon hybridisation assay (ELAHA) using species-specific biotinylated oligonucleotide probes was used to differentiate between JCV and BKV. This assay (VP1-PCR-ELAHA) was evaluated and compared to a PCR assay targeting the human polyomavirus T antigen gene (pol-PCR). DNA sequencing was used to confirm the polyomavirus species identified by the VP1-PCR-ELAHA and to determine the subtype of each JCV isolate. A total of 297 urine specimens were tested and human polyomavirus was detected in 105 specimens (35.4%) by both PCR assays. The differentiation of JCV and BKV by the VP1-PCR-ELAHA showed good agreement with the results of DNA sequencing. Further, DNA sequencing of the JCV positive specimens showed the most prevalent JCV subtype in our cohort was 2a (27%) followed by 1b (20%), 1a (15%), 2c (14%), 4 (14%) and 2b (10%). The results of this study show that the VP1-PCR-ELAHA is a sensitive, specific and rapid method for detecting and differentiating human polyomaviruses JC and BK and is highly suitable for routine use in the clinical laboratory. (C) 2004 Wiley-Liss, Inc.

Identificador

http://espace.library.uq.edu.au/view/UQ:69839

Idioma(s)

eng

Publicador

Wiley-Liss

Palavras-Chave #Polyomavirus #Jcv #Bkv #Pcr #Diagnosis #Subtype #Polymerase-chain-reaction #Progressive Multifocal Leukoencephalopathy #United-states #Virus #Urine #Dna #Recipients #Individuals #Infections #Virology #C1 #270303 Virology #730101 Infectious diseases
Tipo

Journal Article