Multiplex real–time RT–PCR for detecting chikungunya virus and dengue virus

OBJECTIVE: To develop diagnostic test for detection chikungunya virus (CHIKV and Dengue virus (DENV) infection. METHODS: We have performed a rapid, accurate laboratory confirmative method to simultaneously detect, quantify and differentiate CHIKV and DENV infection by single-step multiplex real-time RT-PCR. RESULTS: The assay's sensitivity was 97.65%, specificity was 92.59% and accuracy was 95.82% when compared to conventional RT-PCR. Additionally, there was no cross-reaction between CHIKV, DENV, Japanese encephalitis virus, hepatitis C, hepatitis A or hepatitis E virus. CONCLUSIONS: This rapid and reliable assay provides a means for simultaneous early diagnosis of CHIKV and DENV in a single-step reaction.