An indirect immunofluorescence method for detection of infectious BK virus in urine

Knowles, Wendy A.; Woodroof, M.; Porter, Allison; Gardner, S. D.

doi:10.1016/0166-0934(89)90118-3

Cited by 5 publications

(2 citation statements)

References 5 publications

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“…Traditional methods for detecting and identifying polyomaviruses include serologic methods, virus isolation by cell culture and electron microscopy [Giraldo et al, 1982;Knowles et al, 1989;Nilsen et al, 1991]. Recently, several studies have shown PCR to be an effective tool for detecting polyomaviruses in a range of clinical samples Chang et al, 1996;De Santis and Azzi, 1996]; as a result, PCR is now emerging as the ''gold standard'' for polyomavirus detection.…”

Section: Introductionmentioning

confidence: 99%

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

Whiley

Arden

Mackay

et al. 2004

Journal of Medical Virology

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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.

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Section: Introductionmentioning

confidence: 99%

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

Whiley

Arden

Mackay

et al. 2004

Journal of Medical Virology

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“…Traditional methods for detecting and identifying polyomaviruses include serologic methods, virus isolation by cell culturing, and electron microscopy (15,17,21). Recently, several studies have shown PCR to be an effective tool for detecting polyomaviruses in a range of clinical samples (4,7,11); as a result, PCR is now emerging as the "gold standard" for polyomavirus detection.…”

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confidence: 99%

Detection and Differentiation of Human Polyomaviruses JC and BK by LightCycler PCR

2001

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Human polyomaviruses JC and BK may cause several clinical manifestations in immunocompromised hosts, including progressive multifocal leukoencephalopathy and hemorrhagic cystitis. Molecular detection by PCR is recognized as a sensitive and specific method for detecting human polyomaviruses in clinical samples. In this study, a real-time PCR assay using the LightCycler platform was evaluated and compared to an "in-house" PCR assay using a conventional detection method. A total of 122 urine specimens were tested, and human polyomavirus was detected in 49 specimens (40%) by both conventional PCR and LightCycler PCR. The remaining 73 specimens (60%) were found negative by both assays. For 46 of the 49 positive specimens, LightCycler PCR and conventional PCR identified the same polyomavirus type. These samples included 30 samples with JC virus (JCV), 14 samples with BK virus (BKV), and 2 samples in which both viruses were detected. In the remaining three samples, both JCV and BKV were detected by the conventional assay, but only JCV was detected by the LightCycler assay. The results of this study show that the LightCycler PCR assay displays sensitivity and specificity similar to those of a conventional PCR assay. These data, combined with its rapid turnaround time for results and decreased hands-on time, make the LightCycler PCR assay highly suitable for the rapid detection and differentiation of JCV and BKV in the clinical laboratory.

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Nested PCR for detection of BK virus and JC virus DNA

Bogdanović

Brytting

Cinque

et al. 1994

Clinical and Diagnostic Virology

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An indirect immunofluorescence method for detection of infectious BK virus in urine

Cited by 5 publications

References 5 publications

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

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

Detection and Differentiation of Human Polyomaviruses JC and BK by LightCycler PCR

Nested PCR for detection of BK virus and JC virus DNA

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