目的 检测人源细胞系中EB病毒(Epstein-Barr virus,EBV)、乙型肝炎病毒(Hepatitis B virus,HBV)、丙型肝炎病毒(Hepatitis C virus,HCV)、人乳头瘤病毒(Human pappilomavirus,HPV)、人免疫缺陷病毒(Human immunodeficiency virus,HIV)、人T淋巴细胞白血病病毒(Human T-cell leukemia virus,HTLV)的感染情况.方法 对中心库藏常用的135株人源细胞系分别提取细胞DNA及RNA,采用PCR方法对细胞中的EBV、HBV、HPV、HIV、HTLV进行检测;采用RT-PCR方法对细胞中的HCV进行检测;利用透射电镜观察细胞中是否有病毒颗粒;对经PCR检测HPV核酸片段阳性的细胞系,再利用特异性引物采用PCR方法检测5种高危型HPV(HPV16、HPV18、HPV31、HPV33、HPV45).结果 经PCR检测,135株细胞系中,有4株细胞系(Daudi、Raji、EB-3、NCI-BL2009)中检测到EBV核酸片段;2株细胞系(Hep3B、PLC/PRF/5)中检测到HBV核酸片段;7株细胞系(HeLa、HeLa229、H1HeLa、HeLaS3、SiHa、Ca Ski、CNE-2Z)中检测到HPV核酸片段,其中2株(SiHa、Ca Ski)基因型为HPV16,5株(HeLa、HeLa 229、H1HeLa、HeLaS3、CNE-2Z)基因型为HPV18;1株细胞系(HUT 102)中检测到HTLV核酸片段;没有细胞系中检测到HIV核酸片段;经RT-PCR检测,没有细胞系携带HCV核酸片段.透射电镜下,病毒核酸阳性的细胞系中未观察到病毒颗粒,部分细胞系内可见病毒包涵体类似物.结论 PCR或RT-PCR可以用于检测细胞系中病毒核酸的携带情况;细胞系中病毒核酸以基因组整合的形式存在.%Objective To clarify the virus hosting status of human cell lines.Methods The DNA of Epstein-Barr virus (EBV),Hepatitis B virus (HBV),Human pappilomavirus (HPV),Human immunodeficiency virus (HIV),Human T-cell leukemia virus (HTLV) in 135 human cell lines were checked using PCR,and HCV RNA sequences were checked by RT-PCR.The transmission electron microscopy (TEM) was used to examine the virus particles in cells.The high-risk genotypes of HPV were tested by PCR.Results By PCR assaying,among the 135 human cell lines,four cell lines(Daudi,Raji,EB-3,NCI-BL2009) harbored EBV DNA sequences;two(Hep3B,PLC/PRF/5) harbored HBV DNA sequences;seven (HeLa,HeLa 229,H1HeLa,HeLaS3,SiHa,Caski,CNE-2Z) harbored HPV DNA sequences,including two cell lines (SiHa,Caski) with HPV16,five cell lines (HeLa,HeLa 229,H1HeLa,HeLaS3,CNE-2Z) with HPV18;one cell line(HUT 102) harbored HTLV DNA sequences.No cell line harbored HCV RNA sequences and HIV DNA sequences.No viral particles were observed in the positive cell lines by TEM,but some viral inclusion bodies in certain cell lines.Conclusions The virus hosting status of human cell lines can be checked by PCR or RT-PCR.The viral DNA sequences were integrated in the cellular genome.
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