Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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Application of a PCR Method for the Detection of Mycoplasma in Veterinary Live Viral Vaccines

동물용 생 바이러스 백신에서 Mycoplasma 검출을 위한 PCR 기법 적용

  • Jeon Woo-Jin (National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry) ;
  • Kim Byoung-Han (National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry) ;
  • Jung Byeong-Yeal (National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry) ;
  • An Dong-Jun (National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry) ;
  • Yi Chul-Hyun (National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry) ;
  • Jang Hwan (National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry) ;
  • Chung Gab-Soo (National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry)
  • 전우진 (농림부 국립수의과학검역원) ;
  • 김병한 (농림부 국립수의과학검역원) ;
  • 정병열 (농림부 국립수의과학검역원) ;
  • 안동준 (농림부 국립수의과학검역원) ;
  • 이철현 (농림부 국립수의과학검역원) ;
  • 장환 (농림부 국립수의과학검역원) ;
  • 정갑수 (농림부 국립수의과학검역원)
  • Published : 2005.12.01
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Abstract

We evaluated the PCR assay and two commercialized PCR kits for the detection of mycoplasma in veterinary via live vaccines. The PCR assay could specifically detect all the tested Mycoplasma spp. and Acholeplasma spp., whereas two commercialized PCR kits did not. Also, the specificity of the PCR assay showed that 4 reference strains and 7 field isolates belonging to avian mycoplasma species could be all detected. The sensitivity of the PCR assay was determined using pure cultured Mycoplasma spp. and Acholeplasma spp. with a range of 1 to 100 colony forming units/ml in 9 CFR Mycoplasma broth. To test the availability of the PCR assay for veterinary live viral vaccines, A. laidlawii was artificially inoculated into the swine transmissible gastroenteritis-rota virus combined vaccine and canine parvovirus vaccine, respectively and the sensitivity of the PCR assay was similar with the result of cultured samples. In this study, the PCR assays could be used as rapid and sensitive methods for the detection of mycoplasma in veterinary live viral vaccines.

동물용 생 바이러스 백신 내에 mycoplasma를 검출하기 위해 polymerase chain reaction (PCR)기법과 2가지의 상품화된 PCR 검출킷트를 평가하였다. PCR기법은 시험에 사용된 모든 mycoplasma를 특이적으로 검출할 수 있었으나, 2가지의 상품화된 PCR 검출킷트는 일부의 mycoplasma를 검출하지 못하였다. 또한, PCR기법의 검출 특이도는 조류 유래 mycoplasma에 속한 4주의 표준주 및 7주의 야외분리주를 모두 검출할 수 있었다. PCR기법의 민감도는 9 CFR Mycoplasma액체배지에서 배양한 Mycoplasma 속균 및 Acholeplasma속균에 대해 $1\~100$ colony forming units/ml까지 검출할 수 있었다. 동물용생 바이러스 백신에 대해 PCR기법의 적용가능성을 평가하기 위해, 돼지 전염성위장염 및 로타바이러스 흔합백신과 개 파보바이러스 백신내에 A. laidlawii를 인공적으로 접종한 후, PCR기법의 민감도를 조사하였을 때 배양액을 이용한 검출한계와 유사하였다. 본 연구에서 사용된 PCR 기법은 동물용 생 바이러스 백신내의 mycoplasma를 신속하고 민감하게 검출할 수 있을 것으로 판단되었다.

Keywords

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