A rapid detection of Salmonella species using polymerization chain reaction and Southern hybridization

Polymerization chain reaction과 Southern hybridization을 이용한 Salmonella속 균의 신속한 검출

  • Kim, Won-yong (Korean Collection for Type Cultures Research Institute of Bioscience and Biotechnology, KIST) ;
  • Chang, Young-hyo (Korean Collection for Type Cultures Research Institute of Bioscience and Biotechnology, KIST) ;
  • Park, Kyoung-yoon (Bayer Korea Ltd. Animal Health Division) ;
  • Kim, Chul-joong (College of Veterinary Medicine, Chungnam National University) ;
  • Shin, Kwang-soon (College of Veterinary Medicine, Chungnam National University) ;
  • Park, Yong-ha (Korean Collection for Type Cultures Research Institute of Bioscience and Biotechnology, KIST)
  • 김원용 (한국과학기술연구원 생명공학연구소 유전자은행) ;
  • 장영효 (한국과학기술연구원 생명공학연구소 유전자은행) ;
  • 박경윤 (바이엘코리아(주) 동물의약품사업부) ;
  • 김철중 (충남대학교 수의과대학) ;
  • 신광순 (충남대학교 수의과대학) ;
  • 박용하 (한국과학기술연구원 생명공학연구소 유전자은행)
  • Received : 1995.05.04
  • Published : 1995.07.31

Abstract

Salmonella species are the most prevalent etiologic agents of food-borne acute gastroenteritis. Direct isolation of bacteria from the contaminated food, stool and animal tissues has been used for the diagnosis of salmonellosis routinely. However, isolation of bacteria is time consuming work and not so highly sensitive. In recent years, improved methods of polymerization chain reaction(PCR) and probe hybridization technique have led to the developement of diagnostic assays which employ to detect various human and animal pathogenic bacteria. In this study, we have performed the polymerization chain reaction to detect Salmonella pullorum from tissues and stool samples of chickens with two specific primers, ST5 and ST8C. The target DNA fragment of PhoE gene was successfully amplified from liver, spleen, pancreas, heart, lung, ovary, oviduct and feces samples. The amplified DNA fragments were hybridized with Salmonella typhymurium TA3000 PhoE probe by Southern hybridization. The PCR to amplify the PhoE gene was highly rapid and sensitive method to detect Salmonella pullorum from tissues and stool samples.