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Detection of intestinal bacterial translocation using PCR
T D Kane1, S R Johnson, J W Alexander
1Department of Surgery, University of Cincinnati, Ohio 45267, USA.
The Journal of Surgical Research
|June 1, 1996
Summary
A new polymerase chain reaction (PCR) test can detect bacterial DNA in blood, aiding in the diagnosis of sepsis. This sensitive method helps identify microbial translocation, a suspected cause of organ failure.
Area of Science:
- Microbiology
- Molecular Biology
- Immunology
Background:
- Microbial translocation is implicated in sepsis and multiple organ failure.
- Current diagnostic methods lack sensitivity for sequential detection in humans.
Purpose of the Study:
- To develop a sensitive polymerase chain reaction (PCR) test for detecting Escherichia coli (E. coli) DNA in blood.
- To quantify bacterial translocation from the gut in an animal model.
Main Methods:
- Developed a PCR assay to amplify an 800-bp E. coli gene fragment.
- Achieved high sensitivity (10-100 organisms/0.3 cc blood) using Southern blot detection.
- Utilized a murine model with induced bacterial translocation (gavage) and thermal injury.
Main Results:
- PCR detected E. coli DNA in 100% of culture-positive animals.
- The gavage/burn group showed higher PCR detection rates compared to controls.
- Detectable bacterial DNA was found in some culture-negative animals, indicating translocation.
Conclusions:
- The developed PCR test is highly sensitive and specific for detecting microbial DNA in blood.
- This method allows for repeated testing and potential diagnosis of sepsis of unknown origin.
- It offers a novel approach to monitor microbial translocation in clinical settings.