Fox S A, Finklestone E, Robbins P D, Mastaglia F L, Swanson N R
University Department of Medicine, Queen Elizabeth II Medical Centre, Perth, Australia.
J Neurol Sci. 1994 Aug;125(1):70-6. doi: 10.1016/0022-510x(94)90244-5.
To investigate the hypothesis that the inflammatory muscle diseases (IMD) polymyositis (PM) and dermatomyositis (DM) may be due to a chronic, persistent enterovirus (EV) infection we sought to determine the prevalence of these viruses in muscle tissue using both nested polymerase chain reaction (PCR) and dot-blot hybridization assays. Thirty-six frozen muscle biopsies from 32 adult cases of IMD and 42 biopsies from 36 control subjects with other neuromuscular disorders were studied. Primers for PCR were chosen to conserved regions of the 5'-untranslated region of the EV genome. Constitutive Ableson tyrosine kinase (ABL) mRNA was detected by PCR to confirm the integrity of muscle RNA extracts. The sensitivity of the EV PCR was determined to be 40-400 copies (12.5-125 ag) of synthetic EV RNA transcript against a background of 1 microgram of cellular RNA. The specificity was assessed using a range of enteroviral and unrelated viral isolates extracted from cell cultures. Of the 78 samples tested, ABL mRNA was successfully detected in all but four samples. The time the biopsies spent in ultracold storage (1-73 months) did not appear to influence the integrity of extracted RNA. When assayed for EV RNA by nested PCR, none of 29 IMD cases (i.e., 28 PM and 1 DM) nor sequential biopsies from 3 PM patients were found to be positive. All 42 control biopsies were also negative for EV RNA.(ABSTRACT TRUNCATED AT 250 WORDS)
为了研究炎性肌病(IMD)中的多发性肌炎(PM)和皮肌炎(DM)可能是由慢性持续性肠道病毒(EV)感染所致这一假说,我们试图通过巢式聚合酶链反应(PCR)和点杂交分析来确定这些病毒在肌肉组织中的流行情况。我们研究了来自32例成年IMD患者的36份冷冻肌肉活检样本以及来自36例患有其他神经肌肉疾病的对照受试者的42份活检样本。用于PCR的引物是针对EV基因组5'非翻译区的保守区域选择的。通过PCR检测组成型阿伯尔森酪氨酸激酶(ABL)mRNA以确认肌肉RNA提取物的完整性。测定出EV PCR对合成EV RNA转录本的灵敏度为40 - 400拷贝(12.5 - 125阿克),背景为1微克细胞RNA。使用从细胞培养物中提取的一系列肠道病毒和无关病毒分离株评估特异性。在检测的78个样本中,除4个样本外,所有样本均成功检测到ABL mRNA。活检样本在超低温储存的时间(1 - 73个月)似乎并未影响提取RNA的完整性。通过巢式PCR检测EV RNA时,29例IMD病例(即28例PM和1例DM)以及3例PM患者的连续活检样本均未发现呈阳性。所有42份对照活检样本的EV RNA检测也均为阴性。(摘要截短于250字)