三种提取RNA的方法对汉滩病毒检测敏感性影响的研究

Studies on the sensitivity for the detection of Hantaan virus using defferent RNA extraction methods

分别采用标准法、ＧＰ法和ＭＢＰ法抽提汉滩病毒ＲＮＡ，结合套式ＰＣＲ比较这三种方法的敏感性。结果表明，所用引物对这两株病毒具有相同的特异性。这三种提取ＲＮＡ的敏感性差异不大，敏感性从高到低依次为：ＧＰ、ＭＢＰ和标准法，检测汉滩病毒的效价分别为０．０１、０．０１和０．１ＴＣＩＤ５０／２００μｌ；提取ＲＮＡ的时间长短分别为：标准法为２．５ｈ，ＧＰ为１ｈ，ＭＢＰ为０．５ｈ，可见ＭＢＰ用时最少。使用ＧＰ和ＭＢＰ提取ＲＮＡ时，可以在室温下进行，不需要低温离心。所以，ＭＢＰ最适合于实验室和临床病原体的快速检测。

The sensitivity of extraction RNA by three methods for Hantaan virus detection was studied.Hantaan Virus RNA was extracted by using standard method(guanidium thiocyanate phenol chloroform),glass powder(GP) method and magnetic beads probe(MBP) method.The results shown that the sensitivity of RNA extraction using GP method and MBP method is slightly higher than using standard method.The detection limit of GP method and MBP method was 0.01 TCID 50 /200 μl,that of standard method was 0.1TCID 50 /200 μl.Extraction RNA using standard method,GP method andMBP method taken approximately 2.5h 1h and 0.5h ,respectively.Extraction RNA using GP and MBP may be performed at room temperature and don't need centrifuging at 4℃.So,GP and MBP methods is the most suitable to rapid,sensitive detection of laboratory and clinical pathogens,and specifically suitable to rapid,sensitive detection of sampling environmental samples.