巢式PCR法在日本血吸虫易感水域哨鼠监测中的应用

发布时间：2018-02-19 18:28

本文关键词： 血吸虫日本巢式PCR 哨鼠　出处：《中国病原生物学杂志》2015年04期 　论文类型：期刊论文

【摘要】：目的建立巢式PCR法用以辅助日本血吸虫易感水域中哨鼠的监测。方法选择日本血吸虫基因组重复序列DNA SjR2(2456~3171bp)作为外道PCR的区域,其中2719~3019bp作为内道PCR的扩增区域。以不同稀释度的日本血吸虫成虫基因组DNA为模板进行敏感性测试,并对未感染和感染10条、40条尾蚴的小鼠血清进行检测。将巢式PCR法用于2013年8个哨鼠监测点的监测,观察其应用价值。结果巢式PCR中内道引物扩增产物为301bp,血吸虫基因组DNA浓度低至10-5ng/μl时,仍扩增阳性;以10条和40条尾蚴感染4周的小鼠血清DNA为模板进行巢式PCR,结果为阳性。将巢式PCR用于2013年血吸虫易感水域小范围哨鼠的监测,第2组的20只小鼠中有2只阳性,但解剖小鼠未检出虫体。结论巢式PCR法可辅助血吸虫易感水域中哨鼠的监测,特别对感染度较低水域的监测有一定帮助。
[Abstract]:Objective to establish a nested PCR method to assist sentinel surveillance in susceptible waters of schistosomiasis japonicum. Methods the genomic repeat sequence of Schistosoma japonicum DNA SjR2C24563171bp) was selected as the outer channel PCR region. Among them, 2719 / 3019bp was used as the amplified region of internal PCR. The genomic DNA of Schistosoma japonicum with different dilution was used as a template for sensitivity test. The sera of 10 uninfected and infected mice with 40 cercariae were detected. The nested PCR method was used to monitor 8 sentinel mouse surveillance sites in 2013. Results the PCR products of nested PCR were 301 BP and the genomic DNA concentration of Schistosoma japonicum was as low as 10 ~ (-5) ng / 渭 l. Using 10 cercariae and 40 cercariae infected mice serum DNA as a template, the results were positive. In 2013, the nested PCR was used to monitor the sentinel mice in the susceptible area of schistosomiasis, 2 of the 20 mice in the second group were positive. Conclusion the nested PCR method can assist the surveillance of sentinel rats in susceptible areas of schistosomiasis, especially in the low infection areas.
【作者单位】：南京医科大学病原生物学系;卫生部重要寄生虫病预防与控制高技术重点实验室江苏省血吸虫病防治研究所;
【基金】：国家十一五科技重大专项(No.2008ZX10004-011) 国家十二五科技重大专项(No.2012ZX10004220)
【分类号】：R532.21

【参考文献】