实验性柯萨奇B_(3m)病毒性心肌病变及其病原的原位逆转录-聚合酶链反应检测  

Pathogen detection in the myocardial lesion using in situ RT-PCR in mice induced by experimental CoxB_(3m) virus infection

作　　者：陈曙霞[1] 谢龙山[1] 石迪明[1] 徐欣晖[1] 钱富荣[1] 陈美芳[1] 

机构地区：[1]上海第二医科大学附属仁济医院心内科,200001

出　　处：《中华病理学杂志》2001年第1期46-49,共4页Chinese Journal of Pathology

基　　金：国家自然科学基金资助项目 ( 3 95 70 3 14 )

摘　　要：目的　建立柯萨奇B3m(CoxB3m)病毒性心肌病变模型 ,用原位PCR方法检测心肌病变中的病原 ,以阐明CoxB3m病毒在发病过程中的作用。方法　 (1)用Balb/C小鼠 30只 ,制作CoxB3m病毒性急性心肌病变模型 ,5 0只制作慢性反复感染心肌病变的动物模型 ;并与 2 5只正常Balb/C小鼠相对照 ;(2 )对病毒反复感染所致慢性心肌病变动物模型的左室侧壁厚度及左室腔的面积大小 ,采用KS40 0型图像分析系统进行图像分析 ;(3)应用碱性磷酸酶联接的抗地高辛抗体标记的核苷酸直接掺入法建立原位PCR检测心肌组织中柯萨奇B3m病毒RNA。结果　不论是在急性或反复感染的慢性心肌病变模型心肌组织中 10 0 %可找到CVB RNA阳性信号 ,慢性病变者经图像分析证明左心腔扩大 ,室壁变薄 ,与正常对照组相比P <0 .0 1～ <0 .0 5。结论　在病变心肌组织中 ,用原位PCR检测都有阳性的CVB RNA信号 ,说明柯萨奇B3m病毒不仅能引起急性病变 ,反复感染者亦可造成慢性病变 ,后者心腔常扩大且室壁变薄。Objective To detect the location of pathogens in myocardium using in situ RT PCR technique in order to study the pathogenetic course of the myocardial lesion induced by CoxB 3m virus infection in mice. Methods (1) Thirty and fifty Balb/c mice were used respectively to establish the acute and chronic CoxB 3m infected models, with another 25 healthy mice as the controls; (2) KS400 image analysis system (Germany) was used to measure the cardiac chamber area and the left ventricular wall thickness of the chronic infected mice and the controls; (3) CoxB 3m virus in myocardial tissue was detected using in situ RT PCR by direct incorporated technique which employed nucleotide labeling by anti digoxin antibody and bonded with alkaline phosphatase (anti dig AKP method). Results Picture analysis indicated that the left ventricular chamber area was enlarged and the left ventricular wall was thinner in the chronic repeated virus infected models than those of the controls. With in situ RT PCR, positive signals for Coxsackie virus B 3m RNA were detected not only in the myocardium of the acute Balb/c mice models but also in the myocardium of the chronic mice models. Conclusion Coxsackie virus B 3m is able to induce pathologic lesions by exhibiting positive CVB RNA signals in both acute and chronic models in mice. In the chronic experimental models, the cardiac chamber is enlarged while the ventricular wall is thinned which demonstrates the association with persistent infection of Coxsackie virus B 3m virus.

关 键 词：逆转录聚合酶链反应 病毒性心肌炎 柯萨奇病毒B组 

分 类 号：R542.21[医药卫生—心血管疾病]