Establishment and Application of a Real- time, Duplex PCR Method for Simultaneous Detection of Mycoplasma hyopneumoniae and Mycoplasma hyorhinis
文献类型： 外文期刊
作者： Wu, Yuzi 1 ; Ishag, Hassan Z. A. 3 ; Hua, Lizhong 1 ; Zhang, Lei 1 ; Liu, Beibei 1 ; Zhang, Zhenzhen 1 ; Wang, Haiyan; 1 ;
作者机构： 1.Jiangsu Acad Agr Sci, Natl Ctr Engn Res Vet Bioprod, Key Lab Vet Biol Engn & Technol, Inst Vet Med,Minist Agr, Nanjing 210014, Jiangsu, Peoples R China
2.Univ KwaZulu Natal, Sch Life Sci, Discipline Microbiol, Private Bag X54001, ZA-4000 Durban, South Africa
3.Univ Nyala, Coll Vet Sci, Nyala, Sudan
关键词： Mycoplasma hyopneumoniae; Mycoplasma hyorhinis; duplex real-time PCR; Swine; Detection
期刊名称：KAFKAS UNIVERSITESI VETERINER FAKULTESI DERGISI （ 影响因子：0.685； 五年影响因子：0.655 ）
ISSN： 1300-6045
年卷期： 2019 年 25 卷 3 期
页码：
收录情况： SCI
摘要： The objective of this study was to develop a TaqMan probe-based, sensitive, specific duplex real-time PCR assay for simultaneous detection of Mycoplasma hyopneumoniae and Mycoplasma hyorhinis. The specific primers and probes, labeled with FAM and Texas Red, respectively, were designed to amplify the p97gene of M. hyopneumoniae and p37gene of M. hyorhinis. The duplex real-time PCR reaction mixtures were established and optimized and the sensitivity, specificity and reproducibility of the assay were assessed. The sensitivity of the duplex real-time PCR was found to be 10 copies/mu L for both M. hyopneumoniae and M. hyorhinis, respectively. There was no cross reaction with other common viral and bacterial pathogens. The concentration of standard coefficient of variation of Ct values was less than 5%, indicating a good reproducibility. Clinical samples (n = 937) were tested by the duplex real-time PCR assay, including broncho-alveolar lavage fluids, nasal swabs, tissues and cell culture supernatant. Duplex real-time PCR for simultaneous detection of M. hyopneumoniae and M. hyorhinis was highly sensitive and can be utilized for diagnosing clinical samples. It is time efficient and economic, thereby providing a new approach to control both M. hyopneumoniae and M. hyorhinis.
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