人细小病毒B19实验诊断技术研究

Study on Technology of Experimental Diagnosis of Human Parvovirus B19.

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.