PCR-based identification of postmortem microbial contaminants--a preliminary study.
Investigation of postmortem blood can reveal the presence of significant ethanol levels. However, in some instances it cannot easily be determined if the source of ethanol is from ingestion or from postmortem endogenous fermentation by contaminating microbes. Described here is a robust polymerase chain reaction (PCR)-based method for detecting the presence of common ethanol producing microbial contaminants in human blood. A set of DNA primers were designed for use in PCR to amplify and detect the genomic DNA from humans and three test microorganisms Escherichia coli, Proteus vulgaris, and Candida albicans. A rapid and reproducible protocol was developed for isolating genomic DNA from mixed human blood-microorganism samples that yields a suitable template for PCR. The organism-specific primer pairs can detect the presence of the target microorganisms in human blood at concentrations as low as 10 colony forming units/mL. The PCR products readily can be detected after agarose gel electrophoresis. This method provides an additional means of rapidly identifying microbial contaminants in postmortem blood samples.
Department of Chemistry and Biochemistry, University of Oklahoma, Norman, USA.
D M
DM
Kupfer
A K
AK
Chaturvedi
D V
DV
Canfield
B A
BA
Roe
D002176:Candida albicans / Q000302:isolation & purification / Q000378:metabolism; D004247:DNA / Q000032:analysis; D004269:DNA, Bacterial / Q000097:blood*; D004926:Escherichia coli / Q000302:isolation & purification / Q000378:metabolism; D000431:Ethanol / Q000097:blood; D005554:Forensic Medicine / Q000379:methods*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D011180:Postmortem Changes; D011514:Proteus vulgaris / Q000302:isolation & purification / Q000378:metabolism
D004269:DNA, Bacterial; D000431:Ethanol; D004247:DNA