PCR-DHPLC检测方法在病毒性出血性败血病诊断中的应用

Application of PCR-DHPLC for detection of haemorrhagic septicaemia

运用RT-PCR与变性高效液相色谱技术(DHPLC)相结合,建立了对病毒性出血性败血症病毒(VHSV)快速检测方法,根据对VHSV病毒N基因保守序列的分析,设计一对特异性引物,将RT-PCR扩增产物经变性高效液相色谱进行快速检测。同时运用荧光RT-PCR方法进行平行检测。结果表明,该方法与荧光RT-PCR检测结果相同,其特异性强,灵敏度较高,检测限为3 pg的病毒核酸模板,PCR-DHPLC技术具有高通量、自动化程度高等优势,是一种快速有效的检测方法。

A rapidly detecting method of the viral haemorrhagic septicaemia（VHSV） was established by us ing denaturing high performance liquid chromatography（DHPLC） combined with PCR.One pair of primer was designed according to highly conserved regions of nucleoprontein gene of VHSV.The detection limit of PCR-DHPLC was 3 pg.The results indicated that PCR-DHPLC was a rapid and sensitive assay in detection of VHSV.