Detection and identification of Mycoplasma species by multiplex PCR in cats

서지정보

ㆍ발행기관 : 충북대학교 동물의학연구소 ㆍ수록지정보 : Journal of Biomedical and Translational Research / 19권 / 1호
ㆍ저자명 : Hyun-A Lee, Mi-Na Moon, Jum-Soon Won, Eun-Kyung Im, Ji-Hyun Yoo, Okjin Kim

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

영어 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.

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