PCR technique for detection of toxigenic Pasteurella multocida in mixed bacterial cultures from pigs

Polymerase chain reaction을 이용한 독소생산성 Pasteurella multocida의 검출

  • Chi, Yongzhe (Department of Veterinary Medicine, Kangwon National University) ;
  • Lee, Dong-seok (Department of Veterinary Medicine, Kangwon National University) ;
  • Han, Jeong-hee (Department of Veterinary Medicine, Kangwon National University) ;
  • Han, Kyung-soo (Boehringer Ingelheim Co.) ;
  • Hahn, Tae-wook (Department of Veterinary Medicine, Kangwon National University)
  • Received : 1999.12.10
  • Published : 2000.03.25

Abstract

Pasteurella multocida is kind of commensal bacteria in the upper respiratory tract of pigs. It is classified toxigenic and nontoxigenic strains based on the production of dermonecrotic toxin. Toxigenic strain is most associated with atrophic rhinitis which brings great economical loss in swine industry. However, toxigenic and nontoxigenic strains do not differ by diagnostic biochemical reaction or morphology. One of recently developed techniques, PCR detects the toxigenic P multocida. Amplification of an 846-nucleotide fragment of toxA gene was developed. The fragment amplified by PCR was detected in P multocida type D not type A. The PCR amplification was as sensitive as it could detect 1 pg of P multocida DNA. We compared the result of the PCR with the enzyme linked immunosorbent assay (ELISA) in a test for 40 swine nasal swabs. All of these isolates were toxin negative based on the ELISA while 2 isolates were detected in the PCR technique. in addition to accuracy, as required for rapid detection from contaminated nasal swabs, toxigenic P multocida was recovered efficiently from contaminated culture without inhibition of the PCR. The results show that the PCR detection of toxigenic P multocida directly form nasal swabs are feasible.