Typing of dengue virus Taqman with double real-time PCR

Objective To establishing a multiplex real-time Taqman PCR method to quickly and correctly identify dengue virus type.Methods According to the gene sequences of the four dengue types from the GenBank,four series of dengue virus type-specified primers and probes were designed.Dengue virus I and III's probes were labelled with FAM-TAMARA,while dengue virus II and IV's probes were labelled with JOE-TAMARA.The reaction condition was optimized.Two multiplex real-time PCR methods were established to identify dengue virus I and II and dengue virus III and IV accordingly.Four type dengue virus RNA,dengue virus negative samples,dengue virus RNA diluted samples were tested to identify the specificity,reproducibility and sensitivity of the method.Results The dengue virus I and II(dengue virus III and IV)multiplex typing real-time PCR method could quicky identify dengue virus type witjh good specificity,reproducibility and sensitivity.Conclusion A multiplex real-time Taqman PCR method were established that can quickly identify dengue virus types.