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 reac tion (PCR), it was found that the viral DNA could be released from uri ne by proteinase K and then amplified by PCR directly, without additio nal treatment such as ultracentrifugation or DNA extraction. Direct PC R amplification of viral DNA from urine was volume limited and 5 mu l of urine appeared to be the optimum amount for direct PCR amplificatio n. When the urine volume was greater than 10 mu l, 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 eliminate s several steps which can cause loss of DNA from the sample.