Development of diagnostic method to detect enterovirus by polymerase chain reaction

开发聚合酶链反应检测肠道病毒的诊断方法

• 批准号: 03557042
• 负责人: MATSUMORI Akira
• 金额: $ 8.06万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-03557042/
• 关键词: Gene amplification; Polymerase chain reaction; Enterovirus; Coxsackievirus; Myocarditi; Cardiomyopathy; Cardiovirus; Picornavirus; ポリメラ-ゼ連鎖反応; コクサッキ-ウイルス

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.

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。

项目成果

TAKEMURA G: "Expression and distribution of atrial natriuretic polypeptide in human hypertrophic ventricle of hypertensive hearts and hearts with hypertrophic cardiomyopathy." Ciculation. 83. 181-190 (1991)

TAKEMURA G：“心房钠尿多肽在高血压心脏和肥厚性心肌病心脏肥厚心室中的表达和分布。”

SATO Y: "Effect of immunosimulant therapy on acute viral myocarditis in an animal model." Am Heart J. 124. 428-434 (1992)

SATO Y：“免疫模拟疗法对动物模型中急性病毒性心肌炎的影响。”

MATSUMORI A: "In Cardiomyopthy Updatd4.Cardiomyopathy:Its metabolic and molecular aspects.Antimyosin monoclonal antibody uptake in myocarditis:Clinical and experimental studies." University of Tokyo Press, 6(219-234) (1991)

松森 A：“心肌病 Updatd4。心肌病：其代谢和分子方面。心肌炎中抗肌球蛋白单克隆抗体的摄取：临床和实验研究。”

Anan R: "Cardiomyopathy update 6.Molecular genetic analyses of familial hypertrophic cardiomyopathy." University of Tokyo Press,

Anan R：“心肌病更新 6。家族性肥厚型心肌病的分子遗传学分析。”

Matsumori A: "In Viral infections of the heart.Experimental studies." Edward Arnold,Kent England,

松森答：“心脏病毒感染。实验研究。”