地高辛配基标记探针在流行性出血热尸检组织中原位分子杂交技术的应用和探讨  

APPLICATION OF IN SITU HYBRIDIZATION WITH DIGOXIGENIN-LABELLED PROBES IN AUTOPTIC TISSUES OF EPIDEMIC HEMORRHAGIC FEVER

作　　者：唐志佼[1] 张波[1] 邢寿富[1] 邱慧玲 

机构地区：[1]湖北医学院病理学教研室,武汉430071 [2]中国预防医学科学院病毒学研究所

出　　处：《中国组织化学与细胞化学杂志》1992年第2期180-183,共4页Chinese Journal of Histochemistry and Cytochemistry

摘　　要：将地高辛配基(Dig)标记的探针应用于流行性出血热(EHF)尸检组织的石蜡切片中,进行原位分子杂交四例。方法要点:(1)采用Dig标记的探针及碱性磷酸酶系统、提高其敏感性;(2)为达到被检核酸的暴露彻底、准确,选择合适的消化酶;(3)选择适当的杂交温度和时间,防止了非特异性背景,并避免了组织脱片。结果表明此方法操作简便、安全、快速、敏感。In situ hybridization with Digoxigenin-labelled probes was used for detection of epidemic hemorrhagic fever virus RNA in 4 cases of epidemic hemorrhagic fever autoptic tissues. Its main points are as follows. (1) the sensibility was enhanced by using the digoxigenin-labelling and alkline phosphatase detecting system; (2) nucleotides were accurately and thoroughly exposed by using appropriate digestive enzyme; (3) nonspecific background staining and tissues falling from the slide were prevented by choosing the appropriate temperature and time of hybridization. The results showed that this method was safe, quick, sensitive and easy to operate.

关 键 词：Dig标记 CDNA探针 流行性出血热 原位分子杂交 

分 类 号：Q[生物学]