RT—PCR方法在小鼠肝炎病毒检测中的初步应用

采用4株小鼠肝炎病毒,MHV1、MHV3、A59和JHM的混合液人工感染SCID小鼠,接种病毒后1、2、5d分别取小鼠的全血、肝、肺、脑、脾和血清,用组织总RNA抽提试剂盒提取各组织的总RNA,然后用RTPCR方法检测小鼠肝炎病毒,并对各反应产物进行平均光密度测定。结果显示,接种后24h即可检测出血液中的小鼠肝炎病毒,病毒在小鼠体内各脏器出现的先后顺序为血、肝、脾、肺、脑,主要在肝脏进行复制。

Detection of Mouse Hepatitis Virus in Inoculated SCID Mice by Using Polymerase Chain Reaction

SCID mice were intraperitoneally inoculated with a mixture of MHV1,MHV3,A59,JHM. The blood ,liver,lung,brain and spleen samples were taken, 50mg of liver,lung and brain samples,and 15mg of spleen sample were homogenized respectively with a plastic homogenizer in a 1.5ml microtube for extracting total RNA from the tissues. Then, total RNA was reversely transcribed into cDNA,and the cDNA was amplified by PCR. The results indicated that MHV in blood,liver and spleen was detected 1 day after the inoculation,then in lung and in brain on 2 and 5 days after the inoculation,respectively,and MHV replicated mainly in liver.