建立人微小病毒B19的PCR检测方法

ESTABLISHMENT OF PCR FOR DETECTING HUMAN PARVOVIRUS B19

人微小病毒B19(Human parvovirus B19)是微小病毒科中唯一能感染人的病毒。能够提供B19抗原的病毒体外培养系统尚未建成,因此限制了血清学实验的开展和普及。为此作者建立起B19病毒的PCR检测方法。设计引物在表达外壳蛋白VP1基因区,扩增长度为400bp。对设计引物做了敏感度、特异性和产物鉴定实验,结果显示:1fg B19DAN可被检出;只对B19 DNA有扩增产物出现,而对人基因组DNA,HBV,HPV,HSV,HCMV和ADV DNA均无扩增产物;扩增产物经HaeⅢ,PstⅠ和StyⅠ酶切证实为设计的靶片段。建立的PCR B19 DNA检测方法具有高度的敏感性和特异性,为我国开展B19病毒方面的研究工作打下了基础。

Of several parvoviruses.only the human parvovirus B 19 has been shown to be a human pathogen. Culture systems producing human parvovirus B19 have not been developed in vitro. Consequently,sero diagnostic tests for this infection are not widely available. So we have tried to use PCR technique to delect B19. The designed primers were in the con servative sequences which express the VP1 capsid protein. The experiments of sensitivity .specificity and identification of the amplified products have been carried out for this pair of primers. The results showed that: lfg B19 DN A could be detected ;only amplified products of BI9 could be viewed under UV light,and none of amplifited products of human genome,HBV ,HPV ,HSV ,HCMV ,and ADV could be found. The amplifited products have been identified to be the target DNA ,using Hae III I ,Pst I and Sty I digestions. This method was highly sensitive and specific. It has laid a foundation for investigation of B19 in China.