Giacca M, Severini G M, Mestroni L, Salvi A, Lardieri G, Falaschi A, Camerini F
International Centre for Genetic Engineering and Biotechnology (UNIDO), Trieste, Italy.
J Am Coll Cardiol. 1994 Oct;24(4):1033-40. doi: 10.1016/0735-1097(94)90866-4.
The purpose of this study was to determine the prevalence of enteroviral infection in the myocardium of patients with idiopathic dilated cardiomyopathy by using a highly sensitive and specific detection technique.
Recent molecular studies have suggested that enteroviral persistence (in particular, coxsackieviruses type B) may underlie idiopathic myocarditis and dilated cardiomyopathy.
The method used to detect enterovirus-specific ribonucleic acids (RNAs) is based on reverse transcription and nested polymerase chain reaction amplification with four pairs of primers from the conserved 5' noncoding region of the enteroviral genome. Several members of the Enterovirus genus are detectable by this assay (coxsackieviruses B1 to B6; polioviruses 1 to 3; echoviruses 9, 19 and 31), with a sensitivity threshold close to the detection of a single molecule of viral RNA in 1 mg of tissue sample. Endomyocardial tissue samples from 84 subjects were analyzed (77 samples obtained from left endomyocardial biopsies, 7 from explanted hearts). The subjects comprised 63 study patients (53 with dilated cardiomyopathy, 3 with idiopathic myocarditis, 1 with right ventricular dysplasia, 1 with restrictive cardiomyopathy, 1 with eosinophilic myocarditis, 1 with primary ventricular fibrillation and 3 with myocarditis of known etiology) and 21 control subjects with other diseases.
Positive signals were obtained only in samples from six study patients (four with dilated cardiomyopathy, one with right ventricular dysplasia and one with myocarditis). Samples from control subjects, uninfected rat myocardium and cultured cell lines yielded systematically negative results. Moreover, the nucleotide sequence analysis of the amplification products from patients with positive samples raised doubts about the true positivity of these samples.
This study suggests that the persistence of enteroviral RNA in dilated cardiomyopathy is not a major cause of the disease and that a careful analysis of polymerase chain reaction amplification products is essential in any study in which this technique is pushed to high sensitivity thresholds.
本研究旨在运用高灵敏度和特异性的检测技术，确定特发性扩张型心肌病患者心肌中肠道病毒感染的患病率。
近期分子研究表明，肠道病毒持续存在（尤其是B型柯萨奇病毒）可能是特发性心肌炎和扩张型心肌病的病因。
用于检测肠道病毒特异性核糖核酸（RNA）的方法基于逆转录和巢式聚合酶链反应扩增，使用来自肠道病毒基因组保守5'非编码区的四对引物。该检测方法可检测肠道病毒属的多个成员（B1至B6型柯萨奇病毒；1至3型脊髓灰质炎病毒；9、19和31型埃可病毒），灵敏度阈值接近在1毫克组织样本中检测到单个病毒RNA分子。对84名受试者的心肌内膜组织样本进行了分析（77份样本取自左心室心肌内膜活检，7份取自移植心脏）。受试者包括63名研究患者（53例扩张型心肌病、3例特发性心肌炎、1例右心室发育不良、1例限制性心肌病、1例嗜酸性粒细胞性心肌炎、1例原发性心室颤动和3例已知病因的心肌炎）和21名患有其他疾病的对照受试者。
仅在6名研究患者的样本中获得了阳性信号（4例扩张型心肌病、1例右心室发育不良和1例心肌炎）。对照受试者、未感染大鼠心肌和培养细胞系的样本均系统地产生阴性结果。此外，对阳性样本患者扩增产物的核苷酸序列分析对这些样本的真正阳性结果提出了疑问。
本研究表明，扩张型心肌病中肠道病毒RNA的持续存在不是该疾病的主要病因，并且在任何将该技术推向高灵敏度阈值的研究中，对聚合酶链反应扩增产物进行仔细分析至关重要。
