Development of diagnostic method to detect enterovirus by polymerase chain reaction

建立了用聚合酶链式反应(PCR)检测肠道病毒和心脏病毒的新方法。我们使用了一对包含柯萨奇病毒B3基因组5‘端部分序列的引物，还可以检测到许多肠道RNA。结果发现，感染柯萨奇病毒的C3H/He小鼠在接种病毒后2~21d的心脏组织中病毒RNA呈阳性反应，而在感染病毒后28d时呈阴性反应。检测病毒RNA的时间晚于培养法。因此，酶扩增法检测病毒RNA比常规培养法更快速、更简便，对病毒性心肌炎的临床诊断可能更有价值。我们还对33例心肌炎和心肌病患者的标本进行了研究，在1例心肌病患者中检测到柯萨奇病毒RNA。我们还开发了一种新的检测心脏病毒RNA的方法。在感染了脑心肌炎病毒的小鼠的心脏中，病毒接种后心脏病毒RNA持续存在很长时间。聚合酶链式反应是检测特发性心肌病或扩张型心肌病可能的病毒病因的一种潜在有用的方法。

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.