Detection of BK virus in urine by polymerase chain reaction: a comparison of DNA extraction methods.
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.
Behzadbehbahani
A
A
Department of Pathological Sciences, University of Manchester, Manchester Royal Infirmary, UK.
Klapper
P E
PE
Vallely
P J
PJ
Cleator
G M
GM
D001739:BK Virus / Q000235:genetics* / Q000302:isolation & purification*; D004279:DNA, Viral / Q000302:isolation & purification*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D014508:Urea / Q000737:chemistry; D014556:Urine / Q000737:chemistry / Q000821:virology*
D004279:DNA, Viral; D014508:Urea