Detection of BK virus in urine by polymerase chain reaction: A comparison of DNA extraction methods

A. Behzadbehbahani; P. E. Klapper; P. J. Vallely; G. M. Cleator

doi:10.1016/S0166-0934(97)00101-8

Detection of BK virus in urine by polymerase chain reaction: A comparison of DNA extraction methods

A. Behzadbehbahani, P. E. Klapper, P. J. Vallely^*, G. M. Cleator

^*Corresponding author for this work

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Abstract

Using specimens spiked with BK virus, several DNA extraction methods were evaluated for their ability to remove polymerase chain reaction (PCR) inhibitors from urine samples. It was found that PCR inhibition could be completely overcome by extracting samples with 30% polyethylene glycol (PEG) in 3 M sodium chloride, and partially overcome by extracting samples with guanidine thiocyanate in the presence of high salt concentrations. The nature of the sample inhibition was investigated, leading to the conclusion that both urea and unidentified non-proteinaceous DNA associated substances inhibit BKV DNA amplification from urine.

Original language	English
Pages (from-to)	161-166
Number of pages	6
Journal	Journal of Virological Methods
Volume	67
Issue number	2
DOIs	https://doi.org/10.1016/S0166-0934(97)00101-8
Publication status	Published - 1 Sept 1997

Keywords

BKV
DNA extraction methods
Guanidine thiocyanate
Inhibitors
PCR
Polyethylene glycol
Urine

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10.1016/S0166-0934(97)00101-8

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title = "Detection of BK virus in urine by polymerase chain reaction: A comparison of DNA extraction methods",

abstract = "Using specimens spiked with BK virus, several DNA extraction methods were evaluated for their ability to remove polymerase chain reaction (PCR) inhibitors from urine samples. It was found that PCR inhibition could be completely overcome by extracting samples with 30% polyethylene glycol (PEG) in 3 M sodium chloride, and partially overcome by extracting samples with guanidine thiocyanate in the presence of high salt concentrations. The nature of the sample inhibition was investigated, leading to the conclusion that both urea and unidentified non-proteinaceous DNA associated substances inhibit BKV DNA amplification from urine.",

keywords = "BKV, DNA extraction methods, Guanidine thiocyanate, Inhibitors, PCR, Polyethylene glycol, Urine",

author = "A. Behzadbehbahani and Klapper, {P. E.} and Vallely, {P. J.} and Cleator, {G. M.}",

year = "1997",

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T1 - Detection of BK virus in urine by polymerase chain reaction

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AU - Behzadbehbahani, A.

AU - Klapper, P. E.

AU - Vallely, P. J.

AU - Cleator, G. M.

PY - 1997/9/1

Y1 - 1997/9/1

N2 - Using specimens spiked with BK virus, several DNA extraction methods were evaluated for their ability to remove polymerase chain reaction (PCR) inhibitors from urine samples. It was found that PCR inhibition could be completely overcome by extracting samples with 30% polyethylene glycol (PEG) in 3 M sodium chloride, and partially overcome by extracting samples with guanidine thiocyanate in the presence of high salt concentrations. The nature of the sample inhibition was investigated, leading to the conclusion that both urea and unidentified non-proteinaceous DNA associated substances inhibit BKV DNA amplification from urine.

AB - Using specimens spiked with BK virus, several DNA extraction methods were evaluated for their ability to remove polymerase chain reaction (PCR) inhibitors from urine samples. It was found that PCR inhibition could be completely overcome by extracting samples with 30% polyethylene glycol (PEG) in 3 M sodium chloride, and partially overcome by extracting samples with guanidine thiocyanate in the presence of high salt concentrations. The nature of the sample inhibition was investigated, leading to the conclusion that both urea and unidentified non-proteinaceous DNA associated substances inhibit BKV DNA amplification from urine.

KW - BKV

KW - DNA extraction methods

KW - Guanidine thiocyanate

KW - Inhibitors

KW - PCR

KW - Polyethylene glycol

KW - Urine

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AN - SCOPUS:0030824210

SN - 0166-0934

VL - 67

SP - 161

EP - 166

JO - Journal of Virological Methods

JF - Journal of Virological Methods

IS - 2

ER -

Detection of BK virus in urine by polymerase chain reaction: A comparison of DNA extraction methods

Abstract

Keywords

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