Development of triple antibody sandwich enzyme-linked immunosorbent assays for begomovirus detection

Tomato yellow leaf curl Thailand virus, TYLCTHV, is a begomovirus that causes severe damages to tomato crops in Thailand and other countries across Southeast and East Asia. The development of monoclonal antibodies (MAbs) and serological methods for detecting TYLCTHV is essential for epidemiological studies and screening for virus-resistant cultivars for breeding program.
 
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Researchers developed triple antibody sandwich enzyme-linked immunosorbent assays (TAS-ELISAs) for begomovirus detection using two MAbs (M1 and D2) generated against recombinant TYLCTHV coat protein expressed in E. coli cells. The efficiency of these newly established TAS- ELISAs in detecting TYLCTHV and other begomoviruses was evaluated with tomato, pepper, eggplant, okra and cucurbit plants collected from different provinces of Thailand. TAS-ELISA using the narrow-specificity MAb M1 proved highly efficient for the detection of TYLCTHV and Tobacco leaf curl Yunnan virus (TbLCYnV), whereas TAS-ELISA using the broad-specificity MAb D2 was highly efficient for the detection of TYLCTHV, TbLCYnV, Tomato leaf curl New Delhi virus (ToLCNDV) and Squash leaf curl China virus (SLCCNV). Both newly developed assays allow for sensitive, inexpensive, high-throughput detection of begomoviruses in field plant samples, as well as screening for virus-resistant cultivars.
 
This work was a collaborative study by researchers from BIOTEC Virology and Antibody Technology Research Unit and Kasetsart University.
 
Ref.: Seepiban C, Charoenvilaisiri S, Warin N, Bhunchoth A, Phironrit N, Phuangrat B, Chatchawankanphanich O, Attathom S, Gajanandana O. (2017) Development and application of triple antibody sandwich enzyme-linked immunosorbent assays for begomovirus detection using monoclonal antibodies against Tomato yellow leaf curl Thailand virus. Virology Journal, 14:99.
 

Posted on 20 February 2018.