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用核酸探针鉴定人微小核糖核酸病毒

Identification of human picornaviruses by nucleic acid probes.

作者信息

Hyypiä T

机构信息

Department of Virology, University of Turku, Finland.

出版信息

Mol Cell Probes. 1989 Dec;3(4):329-43. doi: 10.1016/0890-8508(89)90013-3.

Abstract

Human picornaviruses include rhinoviruses and enteroviruses which are responsible for both common and severe clinical diseases. Rhinoviruses are a frequent cause of respiratory infections while members of enterovirus subgroups, polio, coxsackie and ECHO viruses are often responsible for infections of the central nervous system, myocarditis, myositis etc. Human picornaviruses consist of nearly two hundred serotypes and therefore their specific identification after virus isolation, or the diagnosis based on the detection of immune response in patients, is problematic and does not usually provide virological diagnosis at the acute phase of illness. New methods for detection of picornavirus genomic RNA together with increasing knowledge of the nucleotide sequences of this virus group offer interesting possibilities for diagnostic procedures. Spot hybridization, in situ hybridization and enzymatic amplification of specific sequences have successfully been used for this purpose. Probes covering the 5' non-coding part of the genome, and also sequences derived from the region coding for non-structural proteins, can be used as broadly reacting reagents in picornavirus detection. Specific sequences are mainly found in the capsid protein region of the genome. cDNA probes and synthetic oligonucleotides are useful in rapid identification of picornaviruses after amplification in cell cultures and in epidemiological analysis. The biochemical amplification methods may enable recognition of picornaviruses directly in clinical samples in the near future. In situ hybridization methods have been of special interest because they can be used to reveal the presence of enterovirus genomes in biopsy specimens from e.g. affected heart muscle in patients with myocarditis and cardiomyopathy.

摘要

人微小核糖核酸病毒包括鼻病毒和肠道病毒,它们可引发常见及严重的临床疾病。鼻病毒是呼吸道感染的常见病因,而肠道病毒亚组的成员,如脊髓灰质炎病毒、柯萨奇病毒和埃可病毒,常导致中枢神经系统感染、心肌炎、肌炎等。人微小核糖核酸病毒由近200种血清型组成,因此在病毒分离后对其进行特异性鉴定,或基于检测患者免疫反应进行诊断存在问题,且通常无法在疾病急性期提供病毒学诊断。检测微小核糖核酸病毒基因组RNA的新方法,以及对该病毒组核苷酸序列认识的不断增加,为诊断程序提供了有趣的可能性。斑点杂交、原位杂交和特定序列的酶促扩增已成功用于此目的。覆盖基因组5'非编码部分的探针,以及源自非结构蛋白编码区的序列,可作为微小核糖核酸病毒检测中广泛反应的试剂。特定序列主要存在于基因组的衣壳蛋白区域。cDNA探针和合成寡核苷酸可用于在细胞培养物中扩增后快速鉴定微小核糖核酸病毒以及进行流行病学分析。生化扩增方法可能在不久的将来使人们能够直接在临床样本中识别微小核糖核酸病毒。原位杂交方法特别受关注,因为它们可用于揭示肠道病毒基因组在活检标本中的存在,例如心肌炎和心肌病患者受影响心肌的活检标本。

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