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
|
|
1
|
JSP 001
|
ATGTCGAAGCGACCAGGAGAT
|
ACMV DNA-A (CP)
|
783
bp
|
Pita et al., 2001
|
|
JSP 002
|
TGTTTATTAATTGCCAATACT
|
|
2
|
ACMVBF
|
TCGGGAGTGATACATGCGAAGGC
|
ACMV DNA-B (BV1/BC1)
|
628bp
|
Matic et
al., 2012
|
|
ACMVBR
|
GGCTACACCAGCTACCTGAAGCT
|
|
3
|
JSP 001
|
ATGTCGAAGCGACCAGGAGAT
|
EACMV DNA-A (CP)
|
780bp
|
Pita et al., 2001
|
|
JSP 003
|
CCTTTATTAATTTGTCACTGC
|
|
4
|
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
|
|
5
|
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
