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首页> 外文期刊>Parasitology International >Development of a TaqMan based real-time PCR assay for detection of Clonorchis sinensis DNA in human stool samples and fishes
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Development of a TaqMan based real-time PCR assay for detection of Clonorchis sinensis DNA in human stool samples and fishes

机译:基于TaqMan的实时PCR检测试剂盒的开发,用于检测人粪便样品和鱼类中的华支睾吸虫DNA

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摘要

Clonorchiasis caused by the oriental liver fluke Clonorchis sinensis is a fish-borne zoonosis endemic in a number of countries. This article describes the development of a TaqMan based real-time PCR assay for detection of C sinensis DNA in human feces and in fishes. Primers targeting the first internal transcribed spacer (ITS-1) sequence of the fluke were highly specific for C. sinensis, as evidenced by the negative amplification of closely related trematodes in the test with the exception of Opisthorchis viverrini. The detection limit of the assay was 1 pg of purified genomic DNA, 5 EPG (eggs per gram feces) or one metacercaria per gram fish filet. The assay was evaluated by testing 22 human fecal samples and 37 fish tissues microscopically determined beforehand, and the PCR results were highly in agreement with the microscopic results. This real-time PCR assay provides a useful tool for the sensitive detection of C. sinensis DNA in human stool and aquatic samples in China and other endemic countries where O. viverrini and Opisthorchis felineus are absent
机译:由东方肝吸虫引起的支睾吸虫病是许多国家中鱼类传播的人畜共患病。本文介绍了一种基于TaqMan的实时PCR分析方法的开发,该方法可用于检测人粪便和鱼类中的中华绒螯蟹DNA。靶向the虫的第一个内部转录间隔区(ITS-1)序列的引物对中华绒螯蟹具有很高的特异性,在试验中紧密相关的吸虫的负扩增证明了这一点,除了维氏梭菌外。该检测方法的检测限为1 pg纯化的基因组DNA,5 EPG(每克粪便鸡蛋)或每克鱼片1个尾cer。通过检测22份人类粪便样本和37份事先用显微镜确定的鱼组织来评估该测定方法,PCR结果与显微镜结果高度吻合。实时荧光定量PCR为灵敏检测中国及其他缺乏O. viverrini和Opisthorchis felineus的流行国家的人类粪便和水生样品中的中华梭菌DNA提供了有用的工具。

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