卡波肉瘤相关疱疹病毒细胞抗原片的制备  

Preparation of Kaposi′s sarcoma-associated herpesvirus cell antigen tablets

作　　者：文志 周畅 华玮 董凯旋 叶磊 郑豪 徐涵 王林定 Wen Zhi;Zhou Chang;Hua Wei(Dept of Microbiology,Anhui Medical University,Hefei 230032;The Second Clinical College,Anhui Medical University,Hefei 230032)

机构地区：[1]安徽医科大学基础医学院微生物教研室,合肥230032 [2]安徽医科大学第二临床医学院,合肥230032

出　　处：《安徽医科大学学报》2020年第10期1541-1545,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81271837);安徽省自然科学基金(编号:1708085MH193);2019年度安徽医科大学科研基金(编号:2019xkj020)。

摘　　要：目的成功制备了卡波肉瘤相关疱疹病毒(KSHV)的细胞抗原片,并验证其有效性,为检测KSHV血清标本提供了一种可行的办法。方法培养了能够稳定表达卡波肉瘤相关疱疹病毒核相关抗原(LANA)的BCBL-1细胞。BCBL-1细胞是感染过KSHV并且细胞基因组中包含KSHV基因的细胞。待细胞生长活力旺盛且状态良好时,收集细胞并用4%的多聚甲醛固定到预先处理好的抗原片上,完成封闭、待测血清的孵育和荧光二抗的孵育后,通过免疫荧光显微镜观察检测结果。结果通过免疫荧光技术发现,感染过KSHV的患者血清样本能观察到荧光标记的二抗激发的荧光,而未感染过KSHV的标本则没有观察到荧光标记的二抗激发的荧光。结论成功制备了能用于检测血清的卡波肉瘤相关疱疹病毒的细胞抗原片,为进一步研究卡波肉瘤相关疱疹病毒的流行病学提供方法。Objective To prepare Kaposi?s sarcoma-associated herpesvirus(KSHV) cell antigen tablets and verify their effectiveness, providing a feasible method for the detection of KSHV serum samples. Methods BCBL-1 cells stably expressing KSHV nuclear-associated antigen(LANA) were cultured. BCBL-1 cells were cells infected with KSHV and contained the KSHV gene in their genome. LANA is a protein that is highly expressed on the nucleus of a host cell. When the cells were vigorously viable and in good condition, the cells were collected and fixed to the pre-treated antigen tablets with 4% paraformaldehyde. After blocking,incubation of the serum to be tested and incubation of the fluorescent secondary antibody,the results were observed by immunofluorescence microscopy. Results The fluorescence of the fluorescent secondary antibody was observed in the serum samples of patients infected with KSHV by immunofluorescence technique,while the fluorescence of the secondary antibody was not observed by the specimens not infected with KSHV. Conclusion The cell antigenic tablets of KSHV which can detect serum have been successfully prepared,which provides a method for further studying the epidemiology of KSHV.

关 键 词：卡波肉瘤相关疱疹病毒 LANA 细胞抗原片 免疫荧光 

分 类 号：R373.9[医药卫生—病原生物学]