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作 者:林峰[1] 郑美云[1] 曾爱平[1] 杨宁敏[2] 郑敏巧[1] 杨恩[2] 陈弘[1] 饶高峰[1] 张世凡[1] 李敏[1] 吴峰[1] 吴福根[1]
机构地区:[1]浙江省温州医学院附属温岭医院,317500 [2]杭州致远医学检验所,310009
出 处:《医学研究杂志》2007年第2期69-72,共4页Journal of Medical Research
基 金:浙江省卫生厅重点科技项目(2005A111)
摘 要:目的建立一种快速、准确、特异和定量检测人细小病毒B19的实验检测方法。方法以B19病毒NS1基因为靶序列,设计并合成引物和Taqman探针,优化引物与探针比例,调整镁离子浓度,建立对细小病毒B19进行荧光定量检测的实验方法。同时利用荧光定量PCR技术检测本院收集的490例孕妇血清。结果引物与Taqman探针比例为1∶4,镁离子浓度为2.5mmol/L时,反应的本底最低,荧光信号最强。该方法的灵敏度为1.0×101拷贝,有较好的特异性和重复性。检测的490例临床孕妇血清标本,人细小病毒B19DNA阳性为44例,感染率为8.9%(44/490)。结论使用Taqman探针技术的荧光定量聚合酶链反应,能够准确快速定量检测血清中的人细小病毒B19。Objective To establish a rapid specific quantitative assay for human parvovirus B19 detection. Methods Design and synthesize the primers and Taqman-probe which targeted at nonstructural (NSI) protein gene;The fluorescent quantitative PCR was established to detect human parvovirus B19 with the ratio of primer to Taqman probe and the concentration of Mg^2+ regulated, meanwhile, to detected 490 pregnancy serum from hospital( First People' s Hospital of Wenlin ). Results The optimal system of the method was that the ratio of primer to Taqman probe was 1/4 and the concentration of Mg^2+ was 2.5mmol/L. The sensitivity of the assay for Human parvovlrus B19 was 1.0 × 10^1copies,and its specificity and reproduclvity were better than that others copies. Human parvovirus B19 infection rate was 8.9% (44/490) from 490 pregnancy serum by this method. Conclusion This method can rapidly exactly quantitatively detect the Human parvovirus B19.
分 类 号:R373[医药卫生—病原生物学]
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