杨丽;何晓青;韦玉梅;朱轶;程莉;王子健;黄文;

• 1:北京林业大学生物科学与技术学院
• 2:中国科学院生态环境研究中心环境水质学国家重点实验室
• 3:华中农业大学食品科技学院

摘要(Abstract)：

为快速高效地检测人星状病毒,建立了一种新的试验方法——环介导等温扩增技术(loop-mediated isothermal amplification technology,LAMP),并对人星状病毒LAMP反应条件进行了优化。结果表明,镁离子终浓度为4mmol·L-1、甜菜碱终浓度为1mol·L-1的25μL体系下65℃反应90min为其最佳反应条件。对扩增终产物进一步进行酶切分析得到的条带大小与预期的结果(84和135bp)吻合,以人星状病毒、轮状病毒及诺如病毒为模板进行特异性测试亦没有发生交叉反应。将人星状病毒质粒等倍稀释后分别用LAMP及聚合酶链式反应(PCR)检测其灵敏度,结果表明,LAMP与PCR的灵敏度分别达到5.04copies·μL-1和50.4copies·μL-1。用改良的逆转录环介导等温扩增技术(RT-LAMP)及逆转录PCR(RT-PCR)检测12个再生水水样,其中人星状病毒检出率为分别为41.7%(5/12)和33.3%(4/12)。以上结果证明,LAMP是一种检测程序简单、灵敏度和特异性较高的检测手段,在人星状病毒的快速检测方面具有一定的开发潜力。

关键词(KeyWords)： 人星状病毒;环介导等温扩增技术(LAMP);HNB;焦磷酸镁;再生水

Abstract:

Keywords:

基金项目(Foundation): 国家水体污染控制与治理科技重大专项(2009ZX07527-005);; 国家"863"重点项目(2008AA062501);; 中国科学院生态环境研究中心环境水质学国家重点实验室开放基金课题(2009-001)

作者(Author): 杨丽;何晓青;韦玉梅;朱轶;程莉;王子健;黄文;

Email:

DOI:

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