Monoclonal antibody-based serological methods for maize chlorotic mottle virus detection in China

Abstract

Maize chlorotic mottle virus (MCMV) infects maize plants and causes significant losses in corn production worldwide. In this study, purified MCMV particles were used as the immunogen to produce monoclonal antibodies (MAbs) and polyclonal antibodies (PAbs). Four murine MAbs (4B8, 8C11, 6F4, and 9G1) against MCMV were obtained through the hybridoma technology. The triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA), dot-immunobinding assay (DIBA), and immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) using the MAb 4B8 were then developed for sensitive, specific, and rapid detection of MCMV in fields. MCMV could be detected in infected leaf crude extracts at dilutions of 1:327 680, 1:64 000, and 1:3 276 800 (w/v, g/ml) by TAS-ELISA, DIBA, and IC-RT-PCR, respectively. One hundred and sixty-one maize field samples showing virus-like symptoms and sixty-nine symptomless maize field samples from ten different provinces of China were collected and screened for the presence of MCMV using the established serological methods. A phylogenetic tree was constructed based on the full length CP genes and Chinese MCMV isolates formed one branch with Thailand isolates. The detection results demonstrated that MCMV is one of most prevalent viruses infecting maize in the Yunnan and Sichuan provinces of China.

Monoclonal antibody-based serological detection of Citrus yellow vein clearing virus in citrus groves

“…serve the purpose of routine CYVCV detection as well as prevention of the disease spread in groves. Serological detection assays for plant viruses are known to be reliable and cost effective (Shang et al 2011;Wu et al 2011Wu et al , 2013.…”

Further characterization of Maize chlorotic mottle virus and its synergistic interaction with Sugarcane mosaic virus in maize

“…Monoclonal antibodies against MCMV32 and SCMV33, previously produced in our laboratory, were used to analyze leaf samples for the presence of MCMV and SCMV according to the procedure described previously32. MDMV and WSMV were identified using ELISA kits specific for MDMV or WSMV from Agdia (Elkhart, IN, USA) as instructed by the manufacturer.…”

Insights of maize lethal necrotic disease: A major constraint to maize production in East Africa

“…These tests are believed to be the best in identification of large number of field samples (Wu et al, 2013). They are reported as one of the most specific and easiest methods for rapid and precise identification (Naidu et al, 2001;Astier et al, 2007).…”

“…They are reported as one of the most specific and easiest methods for rapid and precise identification (Naidu et al, 2001;Astier et al, 2007). Such tests include enzyme-linked immuno-sorbent assay (ELISA) which includes (triple antibody sandwich ELISA (TAS-ELISA), double antibody sandwich ELISA (DAS-ELISA) and direct antigen coating-ELISA (DAC-ELISA) (Kumar et al, 2004), dot-immunobinding assay (DIBA), and immuno-capture reverse transcription-polymerase chain reaction (IC-RT-PCR) by using the MAb 4B8 that is developed for sensitive, specific, and rapid detection of MCMV in fields (Wu et al, 2013). Other serological tests include; tissue blot immunoassays, immuno-electron microscopy (trapping and decoration), western blots, double immune diffusion and lateral flow rapid tests (Lima et al, 2012).…”