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Detection of multiresistant Salmonella typhimurium DT104 using multiplex and fluorogenic PCR
S A Carlson1, L F Bolton, C E Briggs
1Enteric Diseases and Food Safety Research Unit, National Animal Disease Center, Ames, Iowa 50010, USA. scarlson@nadc.ars.usda.gov
Molecular and Cellular Probes
|June 17, 1999
Summary
Rapid polymerase chain reaction (PCR) assays were developed for the quick detection of multidrug-resistant Salmonella typhimurium DT104. This advancement aids in controlling the spread of this significant public health concern.
Area of Science:
- Microbiology
- Molecular Biology
- Public Health
Background:
- Salmonella infections cause global gastrointestinal and systemic diseases.
- Multidrug-resistant Salmonella typhimurium DT104 is a significant health concern due to antibiotic resistance.
Purpose of the Study:
- To develop rapid, specific, and sensitive diagnostic methods for detecting multidrug-resistant Salmonella typhimurium DT104.
- To facilitate control strategies against this pathogen.
Main Methods:
- Developed a specific polymerase chain reaction (PCR) assay targeting conserved DNA in multidrug-resistant S. typhimurium DT104.
- Utilized multiplex PCR with primers for multiple gene fragments for enhanced specificity.
- Designed a fluorogenic PCR assay for detection in contaminated samples.
Main Results:
- Achieved amplification of specific DNA fragments from multidrug-resistant S. typhimurium DT104 with ACSSuT or ASSuT resistance phenotypes.
- Modified multiplex PCR detected related resistant Salmonella strains like S. typhimurium U302.
- Fluorogenic PCR assay successfully detected target DNA amidst excess contaminating bacterial DNA.
Conclusions:
- Developed a rapid (3-4 hour) PCR-based method for accurate detection of multidrug-resistant S. typhimurium DT104.
- The assay can potentially detect emerging multidrug-resistant Salmonella strains.
- This diagnostic tool can aid in controlling the spread of resistant Salmonella infections.