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PROTOCOL

WAVE UPDATED WORKFLOW FOR PCR TESTING OF SURVEY SAMPLES FOR CASSAVA MOSAIC BEGOMOVIRUSES

Based on results from previous years, WAVE has adopted the following PCR primers (Table  1) and workflow (Figure 1) for the screening of all samples collected during field surveys for  Cassava mosaic begomoviruses (CMB).  

Table 1: Updated WAVE primers for PCR detection of Cassava mosaic begomoviruses

Primer name 

Primer sequences (5′ to 3′) 

Target region 

Size 

Reference 

JSP 001 

ATGTCGAAGCGACCAGGAGAT 

ACMV DNA-A  (CP)

783  

bp

Pita et al.,  2001

JSP 002 

TGTTTATTAATTGCCAATACT 

ACMVBF 

TCGGGAGTGATACATGCGAAGGC 

ACMV DNA-B  (BV1/BC1) 

628bp

Matic et  

al., 2012

ACMVBR 

GGCTACACCAGCTACCTGAAGCT 

JSP 001 

ATGTCGAAGCGACCAGGAGAT 

EACMV DNA-A  (CP) 

780bp

Pita et al.,  2001

JSP 003 

CCTTTATTAATTTGTCACTGC 

CMBRepF 

CRT CAA TGA CGT TGT ACC A 

EACMV DNA-A  (AC1)

650  

bp

Alabi et  

al., 2008

EACMVRepR 

GGT TTG CAG AGA ACT ACA TC 

VNF031/F 

GGATACAGATAGGGTTCCCAC 

EACMV-CM  

DNA-A  

(AC2/AC3)

≈ 

560bp

Fondong  

et al.,  

2000

VNF032/R 

GACGAGGACAAGAATTCCAAT 

6

ACMV21for 

GCAGTGATGAGTTCCCCGGTGCG

DNA-A (AC3- 

AC2-AC1) of  

ACMV, EACMV,  EACMCV,  

EACMKV,  

EACMMV,  

EACMZV and  

SACMV

552  

bp

Matic et  

al., 2012

ACMV21rev 

ATTCCGCTGCGCGGCCATGGAGACC 

 

 

Figure 1 : Workflow for PCR screening of WAVE survey samples for Cassava mosaic begomoviruses