军事医学2016，Vol.40Issue(2)：133-136,4.DOI:10.7644/j.issn.1674-9960.2016.02.014
可视化环介导恒温扩增技术在日本血吸虫感染性钉螺检测中的应用研究
Application of visible loop-mediated isothermal amplification ( LAMP ) technologies in detecting Oncomelania infected with Schistosoma japonicum
摘要
Abstract
Objective To establish a simple and rapid detection technique for Oncomelania infected with Schistosoma japonicum(SJ), with high sensitivity and good specificity .Methods The gene fragment of SJ was amplified by PCR , and cloned into the T-vector to construct positive-reference.An isothermal nucleic acid amplification reaction system for detecting Oncomelania infected with SJ was set up , and its sensitivity was analyzed by detecting positive-reference diluted according to geometric proportion , and its specificity by detecting the genomic DNA of relative samples .Then, a corresponding means of purifying nucleic acid was designed to assemble a reagent detecting Oncomelania infected with SJ . This reagent was validated by detecting Oncomelania samples.Results The 213 bp amplified products were obtained and used to construct recombination T-vector for positive reference .An isothermal nucleic acid amplification reaction system was set up for detecting Oncomelania infected with SJ , and the amplification results could be simply determined by color change, with better sensitivity and specificity .The reagents for detecting Oncomelania infected with SJ were assembled , which could detect samples containing only 1% infected Oncomelania.Conclusion A visible detection method for Oncomelania infected with SJ is successfully established and validated .关键词
日本血吸虫/环境监测/环介导恒温扩增/钉螺Key words
Schistosoma japonicum/environmental monitoring/loop-mediated isothermal amplification/Oncomelania分类
医药卫生引用本文复制引用
冯惊涛,邢微微,孙魁,喻鑫玲,罗志红,毛金武,徐东刚..可视化环介导恒温扩增技术在日本血吸虫感染性钉螺检测中的应用研究[J].军事医学,2016,40(2):133-136,4.基金项目
国家"十二五"科技重大专项资助项目(2012ZX10004909 （）
2013ZX10004101003) （）
