间接核酸杂交方法的建立

建立了一种新的核酸杂交手段一间接核酸杂交方法,其突出的优点是用一种共同的核酸标记物就可检查不同的基因组或不同的基因。我们重组乙型肝炎病毒或EB病毒的核酸片段于噬菌体M_(13)mp8载体,以此重组的单链DNA为第一夹心层,用~(32)P标记的双链噬菌体DNA作为共同探针,检查乙型肝炎病毒和EB病毒的核酸,获得满意的结果。应用该法进行细胞内的原位杂交,检查细胞内存在的EB病毒基因效果亦佳。

INDIRECT HYBRIDIZATION

A new technigue was developed which overcomes the need to introduce label into each specific hybridization probe.This technique links specific sequence1 to another sequence 2.Sequence 2can conveniently be M13mp8 DNA.This unlabeled probe is used in the first hybridization step and in high concentration which favors fast and complete hybridization.The second probe is homologous to the first one, which in our case is labelled double-stranded M13.The labeled probe can bs universally applied in the second step and can form a network on top of sequence 1 which leads to an amplication.We have used this technique for the detection of fragments of hepatitis B and EB viral DNA , and EBV genome containing HRIK cell smear in situ.with completely specific results.