Development of multilocus single strand conformation polymorphism (MLSSCP) analysis of virulence genes of Listeria monocytogenes and comparison with existing DNA typing methods
| Autor: | Bon Kimura, Hajime Takahashi, Itaru Watanabe, Miki Sato, Satoko Handa-Miya, Tateo Fujii, Kazuo Hisa, Asami Yokoi, Seitaro Goto, Takashi Koda |
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| Rok vydání: | 2007 |
| Předmět: |
Genotype
Virulence Factors Virulence Food Contamination Biology Polymerase Chain Reaction Sensitivity and Specificity Microbiology law.invention Ribotyping Species Specificity law Cluster Analysis Typing Phylogeny Polymorphism Single-Stranded Conformational Polymerase chain reaction Genetics Gene Amplification Single-strand conformation polymorphism General Medicine Listeria monocytogenes Meat Products Food Microbiology Multilocus sequence typing Amplified fragment length polymorphism Food Science |
| Zdroj: | International Journal of Food Microbiology. 118:274-284 |
| ISSN: | 0168-1605 |
| Popis: | Development of rapid and simple typing methods is required for analyzing the distribution and contamination routes of food-borne pathogens. We established a simple typing method for Listeria monocytogenes using MLSSCP (Multilocus Single Strand Conformation Polymorphism) analysis. Four virulence genes, hlyA, iap, actA and inlB were amplified by PCR, digested with endonucleases and applied to gels for SSCP. As banding patterns have been shown to reflect even a single nucleotide difference, this method has a potential discriminatory power comparable to that of sequencing analysis. The 64 strains isolated from five meat processing plants were divided into 18 groups by this MLSSCP. Additionally, clustering obtained with this method showed strong correspondence with phylogenetic lineages I and II, and was achieved with much less expenditure in time and cost than is required for other methods, such as MLST. The validity of the MLSSCP lineage classification was confirmed by PFGE, AFLP and ribotyping results. This newly developed MLSSCP method is suitable when obtaining accurate results quickly and simply is crucial. |
| Databáze: | OpenAIRE |
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