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Detection and identification of Mycoplasma species by multiplex PCR in cats KCI 등재

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충북대학교 동물의학연구소 (Research Institute of Veterinary Medicine, Chungbuk National University)
초록

Mycoplasma (M.) felis and M. canis is related with pneumonia or conjunctivitis in domestic cats and several diseases in a variety of other animals, including lower respiratory tract disease or pleuritis. Polymerase chain reaction (PCR) assays has been reported as an easy and useful method. It could be conducted even on nasal swab samples as a non-invasive rapid testing tools for large numbers of Mycoplasma species. However, PCR assays have to conduct multiple assays because of a lot of Mycoplasma species. Therefore, it need to perform several tests and reveal time consuming procedures. In this study, we developed a sensitive and specific multiplex polymerase chain reaction (PCR) assay that detects simultaneously two species like as M. felis and M. canis. The simultaneous detection of M. felis and M. canis primers were used to differentiate two mycoplasma species. The target DNA fragments were specifically amplified M. felis and M. canis PCR with 16S ribosomal DNA primers. Single and mixed Mycoplasma species DNA templates were submitted to validate the specificity of the multiplex PCR. The corresponding specific DNA products were amplified for each pathogen. The detection limit of the developed multiplex PCR is 102 pg with M. felis and M. canis DNA. Furthermore, the developed multiplex PCR detected successfully M. felis in feline nasal specimens. The multiplex PCR assay provides a novel tool for simultaneous detection and differentiation of M. felis and M. canis in cats.

목차
Introduction
 Meterials and Methods
  균주 및 배양 방법
  Genomic DNA 분리
  Multiplex PCR의 개발
  Multiplex PCR의 특이도 검사
  Multiplex PCR의 민감도 검사
  고양이 임상 샘플 검사
 Results
  Mycoplasma 배양 확인
  Multiplex PCR 개발 확인
  특이도 검사 결과
  민감도 검사 결과
  임상 검체 검사 결과
 Conclusion
 ORCID
 Reference
저자
  • Hyun-A Lee(Center for Animal Resource Development, Wonkwang University)
  • Mi-Na Moon(Center for Animal Resource Development, Wonkwang University)
  • Jum-Soon Won(Center for Animal Resource Development, Wonkwang University)
  • Eun-Kyung Im(Center for Animal Resource Development, Wonkwang University)
  • Ji-Hyun Yoo(Technology Service Division, National Institute of Animal Science, Rural Development Administration)
  • Okjin Kim(Center for Animal Resource Development, Wonkwang University) Corresponding author