MOLECULAR DETECTION OF YELLOW FEVER AND RIFT VALLEY FEVER VIRUS USING MULTIPLEX PCR

Background and Aim:Yellow fever (YFV) and Rift valley fever virus (RFV) are RNA virus belonging to the family of flaviviridae and bunyaviridae, respectively. The gold standard for detection of these viruses is cell culture that is time-consuming, laborious, and costly and need expert support (biosafety level 3) for the diagnosis process. For this reason, molecular detection with synthetic construct is an alternative and smart choice for detection of these viruses.Methods:In this study, we used a synthetic vector, containing M Segment for RFV and polyprotein gene for YFV. After designing specific primers, a multiplex PCR reaction was performed and the reaction products were analyzed by 2% agarose gel electrophoresis stained with ethidium bromide.Results:The gel electrophoresis evaluation for the multiplex PCR reaction products analysis showed clear dual band in the regions of 130 bp and 197 bp confirming M Segment & polyprotein gene, respectively. The sensitivity of this method was about 100 ng of plasmid.Conclusion:Multiplex PCR assay is very simple and sensitive for detection of infectious agents. In conclusion, this multiplex PCR method can be used as a simple diagnostic test for identification of RFV and YFV.

Keywords:

Rift valley virus , Yellow fever virus , Detection , Multiplex PCR , Infection