原代猪肝细胞PERV检测及其意义  

Detection and significance of PERV in primary porcine hepatocytes

作　　者：郭海涛[1] 王英杰[1] 刘鸿凌[1] 王海慧[2] 刘俊[1] 黄艳萍[1] 王宇明[1] 

机构地区：[1]中国人民解放军第三军医大学西南医院全军感染病研究所,重庆市400038 [2]中国人民解放军第三军医大学西南医院内分泌科,重庆市400038

出　　处：《世界华人消化杂志》2004年第1期101-104,共4页World Chinese Journal of Digestology

基　　金：国家自然科学专项基金资助项目 No.30027001~~

摘　　要：目的:探讨猪肝细胞内源性逆转录病毒(PERV)检测方法及意义. 方法:采用体外二步灌流法获取猪肝细胞,使用特异性引物对其前病毒序列进行观察;以未逆转录PCR为对照,采用RT-PCR方法检测原代猪肝细胞PERV携带及释放情况,并用套式RT-PCR对扩增产物进行鉴定. 结果:原代猪肝细胞基因组中可检测到PERV前病毒序列,肝细胞中亦可检测到特异性的PERV RNA序列.在缺乏有丝分裂原刺激的情况下,猪肝细胞普通培养的不同时段均可检测到病毒释放,并可持续至细胞死亡. 结论:中国实验小型猪肝细胞携带PERV病毒序列,猪肝细胞普通培养时可释放该病毒,其感染性及生物安全性尚需进一步研究.AIM: To investigate the significance of laboratory detection for PERV in primary porcine hepatocytes. METHODS: Porcine hepatocytes was isolated and cultured with a two-stage perfusion method. Polymerase chain reaction (PCR) were used to detect PERV previrus sequences and reverse transcription polymerase chain reaction (RT-PCR) assays were used to detect PERV RNA sequences by using specific primers in primary porcine hepatocytes and culture supernate, controlled with no RT-PCR. RESULTS: All these PCR assays gave positive results in tissue and serum samples from 2 HCV patients, 1 rabbit and 1 rat. Observation showed the persistent releasing of PERV in the culture supernate in different time points without the stimulation of mitogen by the established method and could last till cell death. CONCLUSION: The method is rapid, cheap and safe, and it will be helpful to the further study of PERV infection and biosafety. The releasing of PERVs in the culture model demonstrates the existence of PERV security in the bioartificial liver support system.

关 键 词：原代猪肝细胞PERV 检测 RT—PCR 前病毒序列 

分 类 号：R575[医药卫生—消化系统]