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从病人外周血单个核细胞中检测HHV-6:分离培养和基因扩增
引用本文:陈斌,姚堃,周瑶玺,季晓辉.从病人外周血单个核细胞中检测HHV-6:分离培养和基因扩增[J].Virologica Sinica,1996(2).
作者姓名:陈斌  姚堃  周瑶玺  季晓辉
作者单位:南京医科大学微生物学教研室
摘    要:收集婴幼儿急疹及淋巴系统增生性疾病患者外周血单个核细胞进行体外培养,从7例婴幼儿急疹及2例淋巴系统增生性疾病患者中分离出一种病毒,此病毒能在PHA激活的人脐血单个核细胞中传代生长,产生典型CPE:形成气球样巨细胞。电镜下观察,感染细胞中可见直径180nm左右,有包膜,疱疹样病毒颗粒;血清学试验证明分离株与HSV-1,2、HCMV、及EBV无抗原交叉,而与HHV-6GS株间存在抗原一致性;多聚酶链反应表明该分离株HHV-6特异性DNA阳性;综合以上结果,初步认为该分离株为HHV-6。同时还用pCR法对所收集的标本直接检测HHV-6特异性DNA。PCR法与病毒分离法相比较,前者HHV-6检出率为88.8%(16/18).后者为38.9%(7/18)。

关 键 词:人类疱疹病毒6型(HH-6),病毒分离培养,鉴定,多聚酶链反应(PCR)

The Detection of HHV-6 in Peripheral Blood Mononuclear Cells from Patients:Virus Isolation and Gene Amplification
Chen Bin,Yao Kun,JiXiaohui,Zhou Yaoxi.The Detection of HHV-6 in Peripheral Blood Mononuclear Cells from Patients:Virus Isolation and Gene Amplification[J].中国病毒学(英文版),1996(2).
Authors:Chen Bin  Yao Kun  JiXiaohui  Zhou Yaoxi
Abstract:eripheral blood samples from patients with exanthem subitum and lymphoproliferative dis-orders were collected.Mononuclear cells were separated from patients blood and were cultured invitro.As a result,virus strains were isolated from the peripheral blood mononuclaear cells of sevenpatients with exanthem subitum and two patients with lymphoproliferative disorders.The isolatescould propagate in the PHA stimulated umbillical cord blood mononucalear cells and the cytopathiceffects characterized by large ballonlike cells were observed after infection Electronmicroscopy ofinfected cells ultrathin section showed the size of enveloped virus particles is 180 nm in diameterand the particles morphologically resemble herpesgroup virus.Serological test further demonstrated that the isolates lacked immunological cross-reactivity with HSV-1 and 2. HCMV and EBV,butwere shown to be antigenically related to HHV-6 GS strain; HHV-6 specific DNAs were detectedin all the isolate by polymerase chain reaction,All the results confirmed that the isolated virus wasHHV-6. In addition,PCR was also used to detect the HHV-6 DNA directly from all the patientssamples.Two methods for detection of HHV-6 were compared,the results showed: the positiverate was 88.8% by PCR and 38.9%by virus isolation.
Keywords:Human Herpesvirus Type 6 (HHV-6)  Virus Isolation  Identification  Polymerase chain reaction(PCR)  
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