Multiplex PCR for Identification and Detection of Cassava Mosaic Begomoviruses in Togo

Abstract

Cassava mosaic disease (CMD) caused by Cassava Mosaic Begomoviruses (CMBs) is one of the most devastating crop diseases and a major constraint for cassava production. In order to ensure surveillance for epidemic prevention, low-cost diagnostic tools are appropriate for large-scale testing of cassava viruses. Multiplex PCR diagnosis is one approach that can reduce diagnostic costs and delays. A multiplex PCR approach was developed for simultaneous detection of African cassava mosaic virus (ACMV), East African Cassava Mosaic Virus and East African cassava mosaic Cameroon virus (EACMV/CM) in Togo CMD-infected cassava leaves. Three primers pairs were used to target their respective viruses in a single tube PCR. Multiplex PCR detected ACMV, EACMV and EACMV/CM in plant DNA extracts prepared from cassava leaves infected with CMB. The primers amplified 783 bp specific to ACMV, 650 bp specific to EACMV and 560 bp specific to EACMCV/CM in both uniplex and multiplex formats. Multiplex PCR is an excellent tool for the effective control of cassava diseases.