Establishment of a Flurogenic Quantitative Polymerase Chain Reaction Assay for Detection of Kio Hepesvirus

A set of primers and TaqMan probe for Fluorescent Quantitation PCR were designed to detect the conservative sequence of Kio Hepesvirus thymidine kinase gene. The standard recombinant plasmids for TK gene were constructed as reference standard used in absolute quantification assay. The reaction conditions of FQ-PCR were optimized. The quantitative curve was established using the reference standard plasmids. The FQ-PCR method was tested for sensitivity,repeatability and specificity by amplification the DNA of the virus. The results demonstrated that the plasmid for FQ-PCR was favorable. The regression coefficient of the quantitative curve was 0.992. The detection system based on FQ-PCR was rapid,sensitive and steady.