Development of Multiplex Real Time Reverse Transcriptase Polymerase Chain Reaction Detection among Zika Virus, Yellow Fever Virus and Dengue Virus Type 4

Zika virus, Dengue virus and Yellow Fever virus are three important Flaviviruses that are epidemic and endemic in Central Africa, Middle and South America, Southeast Asia and Pacific Islands. Recently a multiplex real time RT-PCR for ZIKV, YFV and DENV has developed by out lab. Since the clinical signs caused by these three viral diseases are similar and there is possibility that people may be co-infected with more than one Flavivirus at the same time, it is important to develop an effective and quick method of early diagnostic in the first week of symptom occurrence. Here, we developed a multiplex TaqMan real time RT-PCR for detecting ZIKV, YFV and DENV-4. The primers and probes proved to be highly specificity for their nucleic acid target corresponding to ZIKV, DENV-4 and YFV virus strains used in this study. The Multiplex real time RT-PCR could detect the corresponding viral RNA with a sensitivity between 10 to 100 copies per reaction.