荧光素标记的庚型肝炎病毒基因探针的制备及应用

The sequences of HGV gene that had been reported were checked and analyzed. After comparison of HGV seqences from different virus strains, a gene fragment of 31 nucleotides (7121-7152 nt) served as a probe was labeled by fluorescein-N₆-dd ATP with terminal transferase. The specificity of HGV probe was tested by dot-blot hybridization with HCV RNA—related virus RNA, C.T DNA, HGV-RNA, and Southern-blot hybridization with RT-PCR product of HGV. Positive results obstained only from the HGV-RNA of positive sera that was confirmed by nested RT-PCR, all others were negative. The sensitivity of fluorescein-labeled probe was examined. The result showed that the fluorescein-labeled probe was able to check≥1.56 pg of target genome. The conformity of HGV-RNA detection using RT-PCR and HGV gene probe was 97.9%. The experiment suggested that fluorescein-labeled probe can be used in clinic detection and epidemic study.

Detection of HGV-RNA by Hybridization with Fluorescein-Labeled Probe