Rapid detection of Mycoplasma mycoides subsp. capri and Mycoplasma capricolum subsp. capripneumoniae using high-resolution melting curve analysis

文献类型： 外文期刊

第一作者： Zhang, Jing-peng

作者： Zhang, Jing-peng;Jiang, Jin-xiu;Lin, Yu-sheng;You, Wei;Hu, Qi-lin;Liu, Zhi-cheng

作者机构：

期刊名称：SCIENTIFIC REPORTS （ 影响因子：4.38； 五年影响因子：5.134 ）

ISSN： 2045-2322

年卷期： 2021 年 11 卷 1 期

页码：

收录情况： SCI

摘要： Mycoplasma capricolumsubsp.subsp. capripneumonia (Mccp) and Mycoplasma mycoidessubsp.sbusp. capri (Mmc) cause caprine pleuropneumonia (CCPP) and mycoplasmal pneumonia in goats and sheep (MPGS), respectively. These diseases cannot be identified on clinical symptoms alone and it is laborious to distinguish them using biochemical methods. It is therefore important to establish a simple, rapid identification method for Mccp and Mmc. Here, we report a high-resolution melting (HRM) curve analysis using specific primers based on the Mmc 95010 strain MLC_0560 and Mccp F38 strain MCCPF38_00984 gene sequences. The method was highly specific with intra- and inter-batch coefficients of variation<1%. The lower limit of detection for Mccp and Mmc was 55 copies/L and 58 copies/mu L, respectively. HRM and fluorescence qPCR results were compared using 106 nasal swabs and 47 lung tissue samples from goats (HRM-qPCR coincidence rate 94.8%; 145/153). Mycoplasma isolation and identification was performed on 30 lung tissue samples and 16 nasal swabs (HRM-culturing coincidence rate 87.0%; 40/46). HRM analysis was more sensitive than fluorescence qPCR and Mycoplasma isolation, indicating the practicality of HRM for accurate and rapid identification of Mccp and Mmc, and diagnosis and epidemiology of CCPP and MPGS.

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