J Bacteriol Virol.  2015 Dec;45(4):314-318. 10.4167/jbv.2015.45.4.314.

Modified Method of Multilocus Sequence Typing (MLST) for Serotyping in Salmonella Species

Affiliations
  • 1Division of Enteric Diseases, Center for Infectious Diseases, Korea National Institute of Health, Osong, Korea. leedy0610@korea.kr

Abstract

Multilocus sequence typing (MLST) of Salmonella is useful method for replacing serotyping using antisera but is limited by difficulties associated with in polymerase chain reaction (PCR). We optimized the PCR reaction, especially annealing temperature and extension time (94degrees C for 2 min; 40 cycles at 94degrees C for 30 sec, 56.8degrees C for 1 min, 72degrees C for 2 min; and 72degrees C for 10 min). The degradation of PCR product by thermostable nucleases was inhibited by using template DNAs treated proteinase K or purified by a commercialized preparation kit. The resulting modified MLST was used as accurate and fast typing method.

Keyword

Multilocus sequence typing (MLST); Salmonella spp.; PCR conditions; Serotyping

MeSH Terms

DNA
Endopeptidase K
Immune Sera
Multilocus Sequence Typing*
Polymerase Chain Reaction
Salmonella*
Serotyping*
DNA
Endopeptidase K
Immune Sera
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