A simple method for detecting human polyomavirus DNA in urine by the polymerase chain reaction

In order to develop a simple and sensitive method for detecting human polyomavirus DNA in the urine of patients by the polymerase chain reaction (PCR), it was found that the viral DNA could be released from urine by proteinase K and then amplified by PCR directly, without additional treatment such a...

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Published in:Journal of virological methods 1996-04, Vol.58 (1), p.131-136
Main Authors: Chang, Deching, Wang, Meilin, Ou, Wei-chih, Tsai, Rong-tai, Fung, Chiung-yau, Hwang, yuh-jean
Format: Article
Language:English
Subjects:
Biological and medical sciences
Dialysis
DNA, Viral - urine
Female
Human viral diseases
Humans
Infectious diseases
Inhibitors
JC Virus - genetics
JC Virus - isolation & purification
JCV
Medical sciences
Polymerase Chain Reaction - methods
polyomavirus
Pregnancy
Sensitivity and Specificity
Viral diseases
Viral diseases of the genital and urinary system
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cited_by cdi_FETCH-LOGICAL-c483t-57c15b68650626c561882ff334416335bbe37b1b1211572215cd1c992e5656e43
cites cdi_FETCH-LOGICAL-c483t-57c15b68650626c561882ff334416335bbe37b1b1211572215cd1c992e5656e43
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container_issue 1
container_start_page 131
container_title Journal of virological methods
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creator Chang, Deching
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Ou, Wei-chih
Tsai, Rong-tai
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Hwang, yuh-jean
description In order to develop a simple and sensitive method for detecting human polyomavirus DNA in the urine of patients by the polymerase chain reaction (PCR), it was found that the viral DNA could be released from urine by proteinase K and then amplified by PCR directly, without additional treatment such as ultracentrifugation or DNA extraction. Direct PCR amplification of viral DNA from urine was volume limited and 5 μl of urine appeared to be the optimum amount for direct PCR amplification. When the urine volume was greater than 10 μl, the results of PCR were inconsistent. However, the urine volume could be increased after dialysis to remove possible inhibitor(s) which may interfere with PCR. Direct PCR amplification of patient urine is convenient and eliminates several steps which can cause loss of DNA from the sample.
doi_str_mv 10.1016/0166-0934(95)02001-2
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source ScienceDirect Journals
subjects Biological and medical sciences
Dialysis
DNA, Viral - urine
Female
Human viral diseases
Humans
Infectious diseases
Inhibitors
JC Virus - genetics
JC Virus - isolation & purification
JCV
Medical sciences
Polymerase Chain Reaction - methods
polyomavirus
Pregnancy
Sensitivity and Specificity
Viral diseases
Viral diseases of the genital and urinary system
title A simple method for detecting human polyomavirus DNA in urine by the polymerase chain reaction
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