Fig 1.
Virus disease symptoms observed in the field.
Mosaic, chlorosis, mottling, upward leaf curling, leaf deformation and necrosis symptoms on tomato (A) and black night shade (B).
Table 1.
List of primers used for screening of ToMV infected samples for any other pathogen.
Table 2.
RT-LAMP primers used in the detection of tomato mosaic virus.
Fig 2.
Amplification plot of real-time RT-LAMP for tomato mosaic virus detection.
The amplification curves signifies tp values of 4.45–6.30 minutes and 5.00–10.00 minutes for CTAB (A) and APEG-extracted RNA (C), respectively. (B) and (D) Melt curve profiles of RT-ToMV LAMP amplicons. NTC refers to the nontemplate control (water control), HC is the healthy control and, Pos ToMV is the positive control.
Fig 3.
Sensitivity determination of RT-LAMP for ToMV and comparison with RT-PCR.
(A) ToMV amplification from purified total RNA extracts by RT-LAMP. Reactions were conducted on RNA extracted from ToMV-infected plant using CTAB and diluted in series from 100 ng/μL to 10−9 ng/μL. The RT-LAMP assay was conducted using a Genie II machine (Optigene Ltd., West Sussex, UK). (B) ToMV amplification by RT-LAMP from serially diluted RNA extracted by APEG. (C) ToMV amplification by RT-PCR from purified total RNA; M-1 kb plus DNA ladder (New England Biolabs, Ipswich, MA, USA); ntc-nontemplate control; pos-positive control.
Fig 4.
Specificity determination of RT-LAMP for ToMV.
(A) Amplification curves and (B) annealing derivatives for tomato mosaic virus. The horizontal lines represent ToBRFV, TMV, TSWV, INSV, ToLCArV, CMV, PSTVd and nontemplate control.
Table 3.
Comparison of RT-LAMP with the APEG extraction method and RT-PCR for the detection of tomato mosaic virus.