文献类型： 外文期刊
作者： Yu, Zhichao 1 ; Zhao, Zhiguo 2 ; Chen, Linjun 2 ; Li, Junyan 3 ; Ju, Xianghong 1 ;
作者机构： 1.Guangdong Ocean Univ, Coll Coastal Agr Sci, Dept Vet Med, Zhanjiang 524088, Guangdong, Peoples R China
2.Technol Ctr, Hohhot 010020, Inner Mongolia, Peoples R China
3.Inner Mongolia Acad Agr & Anim Husb Sci, Hohhot 010031, Peoples R China
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.
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