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

作　　者：Jian-xiang WU Qiang WANG Huan LIU Ya-juan QIAN Yan XIE Xue-ping ZHOU 

机构地区：[1]State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University

出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2013年第7期555-562,共8页浙江大学学报（英文版）B辑（生物医学与生物技术）

基　　金：supported by the National Natural Science Foundation of China (No. 31272015);the Ministry of Education of China (No. 313052);the Zhejiang Provincial Natural Science Foundation of China (No. Z3090039)

摘　　要：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, 1:64000, and 1:3276800 (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.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, 8Cll, 6F4, and 9G1） against MCMV were ob- tained 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 1：327680, 1：64000, and 1：3276800 （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 symptomiess maize field samples from ten different provinces of China were col- lected 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.

关 键 词：Maize chlorotic mottle virus (MCMV) Immunocapture reverse transcription-polymerase chain reaction (IC-RT-PCR) Triple antibody sandwich enzyme-linked immunosorbent assay (TAS-ELISA) Monoclonal antibody (MAb) Dot-immunobinding assay (DIBA) 

分 类 号：S435.131[农业科学—农业昆虫与害虫防治]