应用^(32)P 标记DNA探针检测伪狂犬病病毒的研究

DETECTION OF PSEUDORABIES VIRUS BY ^(32)p LABELED DNA PROBE

本文报道建立了一种以^(32)P 标记的 PRV 全基因组 DNA 和重组质粒PSAG1-19DNA 作为探针,检测感染培养细胞中 PRV-DNA 的斑点杂交法。两种探针均能检测10Pg 的 PRVNDA,而与正常细胞抽提物无同源关系,具有较高的灵敏度和特异性。应用了一种快速、简便的 PRV DNA 抽提法,简化了样品制备的程序,提高了检测的可靠性。比较研究了杂交法中的一些重要条件,包括探针的选用,样品 DNA 的处埋,滤膜的选用等,为检测 PRV 提供了一种灵敏、可靠的方法。

A dot blot hybridisation method was developed to detect pseudorabies.virus DNA in infected cultured cells.Five strains of pseudorabies virus were detected by ^(32)p labeled pseudorabies virus DNA probe and recombinant plasmid DNA probe obtained by melecular cloning.Both probes could detect 10pg nucleic acid in the samples.The nucleic acid hybridisation method was sensitive and specific for diagnosing pscudorabies.