1887

Abstract

Summary

A 317-base pair (bp) fragment of the heat shock protein 90 (HSP 90) gene was amplified by the polymerase chain reaction (PCR) for detection of DNA in clinical specimens. One hundred specimens were examined including swabs (39), urines (36), peritoneal fluid (9), pus (8) and blood or serum (8): 23% gave positive results with routine culture, 31% with extended broth culture and 37% with PCR. The amplified product was identified by hybridisation with a radiolabelled internal probe and their restriction enzyme digest patterns (I, III, RI, I and I), which could be predicted from the known sequence of HSP 90. DNA gave the characteristic 317-bp band and specifically hybridised with restriction enzyme-digested candidal DNA. DNA from other sources intermittently gave multiple faint bands especially in the presence of high concentrations of DNA, but these could be readily distinguished. The method was sensitive to 50 pg of DNA (5 pg with radiolabelled probing) and 100 cfu of

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/content/journal/jmm/10.1099/00222615-39-3-233
1993-09-01
2019-12-14
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