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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