Detection of Salmonella species by polymerase chain reaction

Polymerase chain reaction에 의한 Salmonella 속균의 검출

  • Park, Doo-hee (College of Veterinary Medicine, Seoul National University) ;
  • Kim, Won-yong (Genetic Engineering Research Institute, KIST) ;
  • Kim, Chul-joong (College of Veterinary Medicine, Choongnam National University) ;
  • Mah, Jum-sool (College of Veterinary Medicine, Seoul National University)
  • 박두희 (서울대학교 수의과대학) ;
  • 김원용 (한국과학기술연구원 유전공학연구소) ;
  • 김철중 (충남대학교 수의과대학) ;
  • 마점술 (서울대학교 수의과대학)
  • Received : 1994.01.28
  • Published : 1994.01.31

Abstract

In this study, we try to establish the rapid and specific detection system for Salmonella species. The PhoE gene of Salmonella species was amplified with two specific primers, ST5 and ST8c, using PCR. The probe prepared from the amplified PhoE gene was sequenced and applied for Southern blot analysis. After PCR with ST5 and ST8c primers for PhoE gene, DNA bands of expected size(365bp) from 7 different Salmonella species were detected, but not from 12 enterobacteriaceae and 3 gram positive bacteria. PCR was highly sensitive to detect up to 10fg of purified DNA template and to identify Salmonella species with only 320 heat-lysed bacterial cells. The inhibition of PCR amplification from stool specimen was occurred with 50-fold dilution but disappeared over 100 fold dilution of samples. It was confirmed that the PhoE genes were amplified and cloned with over 97% nacleotide sequence homology of PCR products compared with that of S. typhfmurium LT2. The DNA probe derived from S. typhimurium TA 3,000 showed highly specific and sensitive reaction with PCR products of all tested Salmonella species. These results indicate that PCR was rapid and sensitive detection method for Salmonella species and DNA probe prepared from S. typhimurium TA 3,000 was specific to identify PCR products of different Salmonella species.