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An Allelotyping PCR for Identifying Salmonella enterica serovars Enteritidis, Hadar, Heidelberg, and Typhimurium
Published on: July 22, 2011
A rapid multiplex DNA suspension array method for Salmonella typhimurium subtyping using prophage-related markers
Ning-Xia Fang1, Bixing Huang, Lester Hiley
1Public Health Microbiology Laboratory, Division of Communicable Diseases, Queensland Health Forensic and Scientific Services, 39 Kessels Road, Coopers Plains, Qld 4108, Australia. fangningxia@hotmail.com
Journal of Microbiological Methods
|October 22, 2011
Summary
A new method for Salmonella typhimurium subtyping uses multiplex PCR and Luminex array technology. This approach efficiently differentiates strains for epidemiological investigations, offering a fast and economical alternative.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Salmonella typhimurium subtyping is crucial for epidemiological investigations.
- Existing methods may lack efficiency or throughput for large-scale analysis.
- Phage locus typing offers a basis for differentiating bacterial strains.
Purpose of the Study:
- To develop and validate a novel, high-throughput method for Salmonella typhimurium subtyping.
- To utilize multiplex PCR and Luminex DNA suspension array technology for strain differentiation.
- To assess the discriminatory power and efficiency of the developed subtyping method.
Main Methods:
- Selection of 30 markers from prophages and phage-related AFLP fragments.
- Organization of markers into two multiplex PCR assays (15 markers each).
- Development of a two-group DNA suspension array using Luminex xMAP® technology and flow cytometry.
Main Results:
- Application of the method to 438 S. typhimurium isolates representing 56 phage types.
- Generation of 81 distinct profiles, categorizing isolates into 16 main prophage marker profile types.
- High correlation observed between phage types and generated profiles, with a Simpson's Index of Diversity (D) of 0.954.
Conclusions:
- The developed multiplex PCR and Luminex array method is a viable alternative for S. typhimurium subtyping.
- The method demonstrates strong discriminatory power and a high-throughput format suitable for epidemiological studies.
- The technique is fast, economical, and amenable to semi-automation, enhancing its practical application.
