套式PCR在检测SHIV动物模型中的应用  

Application of Nested-PCR in Detection of Chimeric simian/Human immunodeficiency virus in Animal Infection Model

作　　者：李晓燕[1] 孙晓梅[1] 高家红[1] 代成波[1] 陈瑜[1] 李琦涵[1] 代解杰[1] 

机构地区：[1]中国医学科学院中国协和医科大学医学生物学研究所,云南昆明650118

出　　处：《动物医学进展》2005年第12期67-70,共4页Progress In Veterinary Medicine

基　　金：中国综合性艾滋病研究项目基金资助(CIPRA)

摘　　要：用嵌合体猴/人免疫缺陷病毒接种恒河猴,进行体内连续传代,从第4次传代接种嵌合体猴/人免疫缺陷病毒的2只恒河猴外周血淋巴细胞中提取猴全基因组,针对编码嵌合体猴/人免疫缺陷病毒核心蛋白的gag基因进行引物设计,采用套式PCR对提取的全基因组进行检测。套式PCR产物电泳后得到477 bp目的片段,测序结果与GenBank中的猴免疫缺陷病毒mac239的gag序列基本一致,说明嵌合体猴/人免疫缺陷病毒已整合到恒河猴基因组中。与传统病毒分离方法相比较,套式PCR检测的灵敏度明显高于病毒分离方法,尤其在感染初期和感染后期病毒处于潜伏期或病毒载量低的情况下,套式PCR方法的优越性更是传统病毒分离方法所不能替代的。We inoculated SHIV （Chimeric simian/Human immunodeficieney virus, SHIV）into rhesus monkey, and at virus fourth passage we extracted the whole genome from rhesus monkey＇s whole blood. We designed two pairs of primers that were specific to the highly conserved region of gag gene which codes the SIV core protein. Then we used nested-PCR amplification to detect the aimed segment. The nested-PCR products were separated by electrophoresis and we observed the clear aimed 477 bp segments. The sequence result is coincident with the gag sequence of SIV mac 239 in GenBank . This indicated that SHIV had been inoculated into the rhesus monkey. In this article we compare nested-PCR method with the traditional virus isolation method. The results show that the nested-PCR is more sensitive than the traditional virus isolation method especially when the viruses are in incubation periods or when the expression of viruses are too low.

关 键 词：动物模型 套式PCR嵌合体猴/人免疫缺陷病毒 

分 类 号：R512.91[医药卫生—内科学]