Olive D M, Al-Mufti S, Al-Mulla W, Khan M A, Pasca A, Stanway G, Al-Nakib W
Department of Microbiology, Faculty of Medicine, Kuwait University.
J Gen Virol. 1990 Sep;71 ( Pt 9):2141-7. doi: 10.1099/0022-1317-71-9-2141.
A polymerase chain reaction (PCR) assay was used to detect and differentiate picornaviruses (PVs), using primers homologous to the 5' non-coding and VP2 regions of the PV genome. The PCR resulted in a 530 bp PCR product for human rhinoviruses (HRVs) and a 650 bp product for polioviruses, coxsackieviruses (CV) or echoviruses. The PCR assay could detect as little as 1 p.f.u. of virus in either cerebrospinal fluid (CSF) or stool, using ethidium bromide-stained gels. Standard strains of poliovirus, CV, echovirus and HRV were detected, with the exception of echovirus type 22. In contrast, heterologous viruses, such as herpes simplex virus, human cytomegalovirus, adenovirus, influenza virus and rotavirus, as well as human and monkey cell DNA, were not amplified. In nasal swabs taken from patients with respiratory infections, the PCR detected 27 of 28 HRV isolation-positive specimens. All specimens from which viruses other than HRVs were isolated were negative by PCR. The PCR definitively identified poliovirus and CVs from the CSF or stool of patients with aseptic meningitis, as well as CV in the pericardial fluid of a patient who had suffered a myocardial infarction. Specimens taken from patients with similar pathologies, and from which heterologous viruses were isolated, were uniformly negative by PCR.
采用聚合酶链反应（PCR）检测法，使用与微小核糖核酸病毒（PV）基因组5'非编码区和VP2区同源的引物来检测和区分微小核糖核酸病毒（PV）。该PCR反应对人鼻病毒（HRV）产生一个530 bp的PCR产物，对脊髓灰质炎病毒、柯萨奇病毒（CV）或埃可病毒产生一个650 bp的产物。使用溴化乙锭染色凝胶，该PCR检测法在脑脊液（CSF）或粪便中可检测到低至1个感染性病毒颗粒（p.f.u.）的病毒。检测到了脊髓灰质炎病毒、CV、埃可病毒和HRV的标准毒株，但22型埃可病毒除外。相比之下，单纯疱疹病毒、人巨细胞病毒、腺病毒、流感病毒和轮状病毒等异源病毒，以及人和猴细胞DNA均未扩增。在从呼吸道感染患者采集的鼻拭子中，PCR检测到28份HRV分离阳性标本中的27份。所有分离出非HRV病毒的标本经PCR检测均为阴性。PCR明确地从无菌性脑膜炎患者的脑脊液或粪便中鉴定出脊髓灰质炎病毒和CV，以及从一名心肌梗死患者的心包液中鉴定出CV。从患有类似病症且分离出异源病毒的患者采集的标本经PCR检测均为阴性。
