Real-time PCR法用于日本血吸虫感染宿主血清DNA的定量检测及其感染度的评价

发布时间：2018-03-17 04:22

本文选题：日本血吸虫　切入点：TaqMan实时定量PCR　出处：《中国人兽共患病学报》2014年03期 　论文类型：期刊论文

【摘要】：目的以日本血吸虫高度重复序列-逆转录转座子SjR2为靶序列,建立TaqMan实时定量PCR法检测宿主血清中的日本血吸虫DNA用于评价感染度。方法以real-time PCR法检测不同感染度的家兔模型血清DNA。结果该法具有高度的敏感性和特异性,可以检测到44.7拷贝的重组质粒DNA。在感染家兔后的3d到7周血清中均能检测到血吸虫DNA,不同感染度早期检测的时间节点及其检测阈值分别为,家兔感染30条尾蚴(EPG=14)需在感染后2周才能检测到目的DNA,其含量为119.03拷贝,感染50条尾蚴(EPG=24)、感染100条尾蚴(EPG=48)则在感染后1周可检测到目的DNA,其含量分别为54.36拷贝和72.24拷贝,在家兔感染200条尾蚴(EPG=97)、感染500条尾蚴(EPG=232)最早在感染后3d即可检测到目的 DNA,其含量分别为60.34拷贝和142.47拷贝。日本血吸虫感染宿主血清DNA水平与感染度呈正相关,即血清DNA浓度随感染度的增大而上升。结论本研究所建立的real-time PCR法可定量检测血清DNA动态变化并对日本血吸虫病诊断及感染度的评价具有潜在的应用价值,为日本血吸虫病诊断与疗效考核提供了新方法。
[Abstract]:Objective to take the highly repetitive sequence of Schistosoma japonicum reverse transcription transposon SjR2 as the target sequence. A real-time quantitative TaqMan PCR method was established for the detection of schistosomiasis japonicum DNA in host serum. Methods real-time PCR assay was used to detect serum DNA in rabbits with different infection degrees. The results showed that the method was highly sensitive and specific to the infection of Schistosoma japonicum. The DNA of Schistosoma japonicum could be detected in the sera of rabbits from 3 days to 7 weeks after infection. The time node of early detection of different infection degree and the detection threshold were respectively. In rabbits infected with 30 cercariae, the target DNA was detected only 2 weeks after infection. The content of the target DNA was 119.03 copies, 50 cercariae were infected with EPGM24, 100 cercariae were infected with EPG4), and the target DNA was detected 1 week after infection, the contents of which were 54.36 copies and 72.24 copies, respectively. In rabbits infected with 200 cercariae and 500 cercariae, the target DNA was detected at the earliest 3 days after infection, the contents of which were 60.34 copies and 142.47 copies, respectively. The serum DNA level of Schistosoma japonicum infected host was positively correlated with the degree of infection. Conclusion the real-time PCR method established in this study can quantitatively detect the dynamic changes of serum DNA and has potential application value in the diagnosis and infection evaluation of schistosomiasis japonicum. It provides a new method for diagnosis and curative effect assessment of schistosomiasis japonicum.
【作者单位】：苏州大学医学部基础医学与生物科学学院病原生物学系;
【基金】：国家973计划项目子课题(No.2007CB513102)~~
【分类号】：R346

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