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| Establishment of a SYBR Green Ⅰ qRT-PCR for Rapid Detection of Mycoplasma capricolum subsp. capripneumoniae |
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Abstract Mycoplas macapricolum subsp. capripneumoniae (Mccp) is the causative agent of contagious capri pleuropneumonia (CCPP). A SYBR Green I real-time fluorescent quantitative PCR (SYBR Green I qRT-PCR) was established based on primers derived from Mccp strain F38 (No. LN515398.1) arcD (MCCPF38_00114) gene deposited in GenBank. The results showed that the assay could only detect Mccp, and had no cross-reactivity with Mycoplasma mycoides subsp. capri (Mmc), Myplasma ovipenumoniae (Mo), Orf virus (ORFV)，Staphylococcus aureus (SA), Pasteurella multocida (Pm), Escherichia coli (Ec) and Haemophilus parasuis (HPS), indicating a high specificity. The sensitivity of the assay was 279 copies/μL with 1000 times higher than that of the conventional PCR. In addition, the variations in intra- and inter-assays were both less than 1%, indicating a good repeatability. The results indicate that the SYBR Green I qRT-PCR assay could provide a early, rapid and more sensitive diagnostic method for CCPP and a quantitative analysis assay for Mccp.
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Received: 07 June 2017
Published: 04 February 2018
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