DEVELOPMENT OF A DROPLET DIGITAL PCR FOR DETECTION OF TRICHURIASIS IN SHEEP

文献类型： 外文期刊

第一作者： Yu, Zhichao

作者： Yu, Zhichao;Ju, Xianghong;Yu, Zhichao;Zhao, Zhiguo;Chen, Linjun;Li, Junyan

作者机构：

关键词： Trichuriasis; Droplet Digital PCR; Real-Time Quantitative PCR; Trichuris; Sheep Feces; Molecular Diagnostics

期刊名称：JOURNAL OF PARASITOLOGY （ 影响因子：1.276； 五年影响因子：1.486 ）

ISSN： 0022-3395

年卷期： 2020 年 106 卷 5 期

页码：

收录情况： SCI

摘要： Trichuriasis is a serious threat to the economic development of animal husbandry. This research aimed to establish a droplet digital PCR (ddPCR) method to detect Trichuris spp. for the early diagnosis and prevention of trichuriasis in sheep. The real-time quantitative PCR (qPCR) and ddPCR methods were used for the detection of nematodes by targeted amplification of the ITS gene. Each means was evaluated to optimize the limit of detection and reproducibility. For a recombinant plasmid, the qPCR results showed that the detection limit was 31.7 copies per reaction. In contrast to qPCR, ddPCR was able to detect concentrations below 3.17 copies per reaction. Both assays exhibited good reproducibility. However, the ddPCR method was more stable for low-copy-number detection. This new assay was specific for Trichuris spp. and did not cross-react with other relevant gastrointestinal nematodes. A total of 98 clinical samples were tested with both assays. The results showed that the positive rate of ddPCR (80.6%) was higher than that of qPCR (72.4%). This method could be used as an efficient molecular biology tool to test for Trichuris spp. and could be a new valuable tool for the clinical diagnosis and prevention of trichuriasis.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序