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目的建立定位检测细胞或组织中柯萨奇B病毒RNA的原位RT-PCR方法(直接法)。方法对柯萨奇B病毒(1~6型)感染HeLa细胞,经核酸酶(DNase和RNase)处理后的病毒感染HeLa细胞以及非感染的HeLa细胞进行原位RT-PCR检测。结果经扩增后,病毒感染的HeLa细胞呈蓝黑阳性信号;并且这种阳性信号经RNaseA作用而消失,却不受DNase作用的影响;而非感染的HeLa细胞则不着色。结论原位RT-PCR法可特异地检测柯萨奇B病毒感染HeLa细胞中的病毒RNA,是一种特异和敏感的定位检测方法,可应用于研究该病毒的持续性感染及其与克山病等相关疾病的关系
Objective To establish an in situ RT-PCR method to detect Coxsackie B virus RNA in cells or tissues (direct method). Methods HeLa cells infected with Coxsackie B virus (type 1 to 6) and HeLa cells treated with nuclease (DNase and RNase) and non-infected HeLa cells were detected by in situ RT-PCR. Results After amplification, the virus-infected HeLa cells showed a blue-black positive signal; and this positive signal disappeared by RNaseA but was not affected by DNase; while non-infected HeLa cells were not colored. Conclusion The in situ RT-PCR method can detect the viral RNA of Coxsackie B virus infected HeLa cells specifically. It is a specific and sensitive method for localization detection and can be used to study the persistent infection of the virus and its interaction with Keshan Disease and other related diseases