Muir P, Nicholson F, Jhetam M, Neogi S, Banatvala J E
Department of Virology, United Medical School, Guy's Hospital, London, United Kingdom.
J Clin Microbiol. 1993 Jan;31(1):31-8. doi: 10.1128/jcm.31.1.31-38.1993.
We describe a rapid method for extraction and detection of enterovirus RNA in clinical samples. By using magnetic bead technology, enterovirus RNA was efficiently and rapidly extracted from cerebrospinal fluid, stool, saliva, blood, pericardial fluid, urine, and cryopreserved or formalin-fixed solid tissue. Enterovirus RNA was then detected by reverse transcription followed by polymerase chain reaction amplification with primers designed to allow detection of most enterovirus serotypes. For detection of enteroviruses in specimens from patients with acute enteroviral disease, the overall sensitivity of enzymatic RNA amplification was greater than that of cell culture isolation, especially in blood specimens and in stool specimens from patients with acute cardiac disease. Enterovirus RNA was also detected in cryopreserved and archival formalin-fixed myocardial tissue from patients with acute myocarditis and chronic dilated cardiomyopathy. The ability to study archival specimens is of particular value in conducting retrospective investigation. The RNA extraction procedure used was considerably faster than extraction methods using organic reagents, used less hazardous reagents, and was of similar sensitivity. This detection protocol may therefore be useful both for the diagnosis of enterovirus infection and in studying the pathogenesis of acute and chronic enterovirus-induced disease.
我们描述了一种从临床样本中提取和检测肠道病毒RNA的快速方法。通过使用磁珠技术，可从脑脊液、粪便、唾液、血液、心包液、尿液以及冷冻保存或福尔马林固定的实体组织中高效快速地提取肠道病毒RNA。然后通过逆转录，接着用设计用于检测大多数肠道病毒血清型的引物进行聚合酶链反应扩增来检测肠道病毒RNA。对于急性肠道病毒疾病患者标本中肠道病毒的检测，酶促RNA扩增的总体灵敏度高于细胞培养分离法，尤其是在急性心脏病患者的血液标本和粪便标本中。在急性心肌炎和慢性扩张型心肌病患者的冷冻保存及存档福尔马林固定心肌组织中也检测到了肠道病毒RNA。研究存档标本的能力在进行回顾性调查中具有特别的价值。所使用的RNA提取程序比使用有机试剂的提取方法快得多，使用的危险试剂更少，且灵敏度相似。因此，这种检测方案可能对肠道病毒感染的诊断以及研究急性和慢性肠道病毒引起疾病的发病机制都有用。
