Development of diagnostic method to detect enterovirus by polymerase chain reaction

利用聚合酶链反应（PCR）技术建立了检测肠道病毒和心脏病毒的新方法。我们使用了一对引物，它包含了柯萨奇病毒B3基因组的5'端序列的一部分，也可以检测许多肠道RNA。用PCR方法检测感染柯萨奇病毒的C_3H/He小鼠心脏，发现病毒感染后2 ~ 21天呈阳性反应，28天后呈阴性反应。PCR检测病毒RNA的时间晚于培养。因此，使用酶扩增检测病毒RNA比常规培养方法更快速、更容易，并且可能对病毒性心肌炎的临床诊断更有用。我们还对33例心肌炎和心肌病患者的标本进行了研究，并在1例心肌病患者中检测到柯萨奇病毒RNA。我们还建立了一种新的方法，用PCR检测心脏病毒RNA。在感染脑心肌炎病毒的小鼠心脏中，病毒接种后很长时间内，病毒RNA仍然存在。PCR是一种潜在的有用的方法来测试可能的病毒病因在特发性心肌病或扩张型心肌病。

New methods to detect enterovirus and cardiovirus were developed using polymerase chain reaction (PCR). We used a pair of primers, which encompass a part of the 5' end sequence of the coxcsackieviurus B3 genome and can also detect many enterovial RNAs. We found that hearts of C3H/He mice infrrted with coxsackievirus were positive for the viral RNA from 2 to 21 days after virus inoculation by PCR, but negative after day 28. The viral RNA were detected by PCR later than by culture. Thus, the detection of the viral RNA using enzymatic amplification is more rapid and easier and may be more useful for clinical diagnosis of viral myocarditis than coventional culture methods. We also studied 33 specimen from the patients with myocarditis and cardiomyopathy and coxsackieviral RNA was detected in a patient with cardiomyopathy. We also developed a new method to detect cardioviral RNA using PCR. Cardioviral RNA persisted long after virus inoculatin in the hearts of mice infected with encephalomyocarditis virus. The PCR is a potentially a useful method to test possible viral etiologies in idiopathic heart muscle disease or dilated cardiomyopathy.