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PCR-based identification of postmortem microbial contaminants--a preliminary study
D M Kupfer1, A K Chaturvedi, D V Canfield
1Department of Chemistry and Biochemistry, University of Oklahoma, Norman, USA.
Journal of Forensic Sciences
|July 17, 1999
Summary
This study presents a new polymerase chain reaction (PCR) method to detect ethanol-producing microbes in postmortem blood. This technique helps distinguish between ethanol from ingestion and microbial fermentation, aiding forensic investigations.
Area of Science:
- Forensic Science
- Microbiology
- Molecular Biology
Background:
- Postmortem blood ethanol analysis is crucial for forensic investigations.
- Distinguishing ethanol from ingestion versus endogenous fermentation by microbes can be challenging.
- Microbial contamination can lead to inaccurate ethanol level interpretations.
Purpose of the Study:
- To develop a reliable method for detecting common ethanol-producing microorganisms in postmortem blood.
- To differentiate microbial contamination from ethanol of exogenous origin.
- To enhance the accuracy of postmortem ethanol analysis.
Main Methods:
- Development of a polymerase chain reaction (PCR)-based assay.
- Design of DNA primers specific for human DNA, Escherichia coli, Proteus vulgaris, and Candida albicans.
- Establishment of a rapid protocol for genomic DNA isolation from mixed samples.
- Detection of PCR products via agarose gel electrophoresis.
Main Results:
- The PCR method successfully amplified genomic DNA from target microorganisms.
- The assay detected microbial contaminants at concentrations as low as 10 colony forming units/mL.
- The protocol is rapid, reproducible, and suitable for forensic applications.
Conclusions:
- The developed PCR method provides a robust tool for identifying microbial contaminants in postmortem blood.
- This technique aids in accurately determining the source of ethanol in forensic toxicology.
- It offers a rapid and sensitive means to resolve ambiguities in postmortem ethanol findings.