摘要
用地高辛配基（Ｄｉｇｏｘｉｇｅｎｉｎ，Ｄｉｇ）、生物素（Ｂｉｏｔｉｎ，Ｂｉｏ）和辣根过氧化物酶（Ｈｏｒｓｅｒａｄｉｓｈｐｅｒｏｘｉｄａｓｅ，ＨＲＰ）分别标记登革病毒２型（Ｄｅｎｇｕｅｖｉｒｕｓｔｙｐｅ２，Ｄｖ２）的ｃＤＮＡ，制备三种非同位素标记探针，与感染ＤＶ２的Ｃ６／３６细胞上清中病毒核酸做斑点杂交。结果表明：三种标记探针均只与ＤＶ２－ＲＮＡ呈强阳性杂交反应，显示ＤＶ２型特异性；Ｄｉｇ－探针最灵敏，可检出ＤＶ２－ＲＮＡ的最小量为０．１ｐｇ，其次为ＨＲＰ－探针（可检出０．３ｐｇ）和Ｂｉｏ－探针（可检出１～３ｐｇ）。用ＰＣＲ技术制备探针并同步标记比其他方法简便、快速，只需少量模板数小时内可获得足够量的标记探针，是值得推广使用的标记探针制备方法。
igoxigenin,biotin and horseradish peroxidase(HRP) were used to prepare
nonisotope labelled DV2-cDNAproberespectively. These probes were used to detect DV-RNA
extracted from DV2 infected supernatant of C6/36cell. The results of cDNA : RNA dot blot
hybridization showed that these nonisotope labelled probes were highlyspecific. The
digoxigenin labelled probe was the most sensitive among the three kinds fo probes and the tiny
amount(0.1pof DV2-RNA was detectable.The amount of 0.3pg and 1-3pg DV2-RNA can be
detected byHRP and bi-otin labelled probes,respectively.Our results suggest that a large
amount of labelled probes can be obtaint using a small quantity of templatewith labelling
method.The preparation of labelled probes with PCR may have great value in producing
labelledprobes.The modified NaI method for extracting DV-RNA was more convenient and much
faster.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
1994年第6期426-429,共4页
Chinese Journal of Microbiology and Immunology
基金
CMB基金
卫生部医学重点科技项目资金
关键词
登革病毒
核酸杂交
地高辛配
探针
生物素
Dengue virus
Nonisotope labelled probes
Nucleic acid hybridization
