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目的:对一株从中国河南省一名艾滋病患者体内分离的、可在MT4细胞上稳定传代的未知病毒29A进行鉴定,确定该病毒种类及型别。方法:用从该患者外周血单核细胞中分离的病毒株感染MT4细胞,获得可在该细胞上稳定传代的病毒株29A。通过细胞病变特征、电镜观察结果、H IV-1p24抗原检测以及H IV-1 pol区基因扩增结果,对该毒株是否为H IV-1进行鉴别。在电镜下对病毒感染细胞的超薄切片进行观察,发现该病毒呈现疱疹病毒形态特征,因此设计人类疱疹病毒6型(HHV-6)U69、U16/U17、U60/U66三个不同基因片段的特异性引物并进行巢式PCR扩增,对扩增产物进行序列测定和结果分析。结果:该毒株H IV-1 p24抗原检测为阴性;用H IV-1 pol区特异性引物未扩增出目的片段;细胞病变形态不同于H IV-1引起的病变;电镜观察该病毒直径为160~200 nm,明显大于H IV-1,表明该传代株不是H IV-1。用人类疱疹病毒6型(HHV-6)U69、U16/U17、U60/U66等基因的特异性引物均扩增出了目的片段,序列分析的结果表明该毒株为HHV-6病毒B亚型。结论:在国内首次从艾滋病患者的外周血淋巴细胞中分离出HHV-6,为进一步研究HHV-6和H IV-1的相互作用奠定了基础。
OBJECTIVE: To identify a 29A unknown virus that can be stably passaged on a MT4 cell isolated from an AIDS patient in Henan Province of China, and to determine the type and genotype of the virus. Methods: MT4 cells were infected with the virus isolated from peripheral blood mononuclear cells of this patient to obtain strain 29A which can be stably passaged on this cell. The virus was identified as H IV-1 by cytopathic features, electron microscopy, H IV-1 p24 antigen detection and H IV-1 pol gene amplification. Under the electron microscope, the virus-infected cells were observed by ultrathin section and found that the virus showed herpesvirus morphological characteristics. Therefore, three different gene fragments of human herpesvirus type 6 (HHV-6) U69, U16 / U17, U60 / U66 Specific primers and nested PCR amplification of the amplified products were sequenced and the results were analyzed. Results: The H IV-1 p24 antigen was negative for this strain. The specific fragment of H IV-1 pol region was not amplified. The morphological changes of the cells were different from those induced by H IV-1. The diameter of the virus Was 160-200 nm, significantly greater than H IV-1, indicating that the passage strain was not H IV-1. The target fragment was amplified with specific primers of human herpesvirus type 6 (HHV-6) U69, U16 / U17, U60 / U66 and other genes, and the sequence analysis showed that the strain was HHV-6 virus B subtype . Conclusion: HHV-6 was isolated from peripheral blood lymphocytes of AIDS patients in China for the first time, which laid the foundation for further study on the interaction between HHV-6 and H IV-1.