Monoclonal antibody-based serological methods for maize chlorotic mottle virus detection in China
Monoclonal antibody-based serological methods for maize chlorotic mottle virus detection in China
复制标题
基于单克隆抗体的中国玉米褪绿斑驳病毒检测血清学方法
DOI：
10.1631/jzus.b1200275
复制
发表时间：
2013-07
影响因子：
5.1
通讯作者：
Zhou, Xue-ping
中科院分区：
文献类型：
--
作者：
Liu, Huan;Qian, Ya-juan;Xie, Yan;Zhou, Xue-ping
玉米褪绿斑驳病毒（MCMV）是一种侵染玉米的病毒，在世界范围内引起玉米生产的重大损失。本研究以纯化的MCMV颗粒为免疫原，制备了MCMV单克隆抗体（MAbs）和多克隆抗体（PAbs）。通过杂交瘤技术获得了4株抗MCMV的单克隆抗体（4 B8、8 C11、6 F4和9 G1）。建立了三抗体夹心酶联免疫吸附试验（TAS-ELISA）、斑点免疫结合试验（迪巴）和免疫捕获逆转录聚合酶链反应（IC-RT-PCR）检测MCMV的方法。TAS-ELISA、迪巴和IC-RT-PCR分别在1：327 680、1：64 000和1：3 276 800（w/v，g/ml）的稀释度下检测到MCMV。从我国10个省（市、区）采集了161份表现病毒样症状的玉米田样品和69份无病毒样玉米田样品，采用已建立的血清学方法对MCMV进行了检测。根据CP基因的全长序列构建了系统发育树，中国MCMV分离物与泰国分离物形成一个分支。检测结果表明，MCMV是云南和四川两省玉米上最流行的病毒之一。
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.
登录
查看更多内容
影响因子：
1.5
作者：
L. Nault;D. T. Gordon;J. Loayza
通讯作者：
L. Nault;D. T. Gordon;J. Loayza
DOI：
--
发表时间：
1978
期刊：
Plant disease reporter
影响因子：
--
作者：
C. Niblett;L. Claflin
通讯作者：
C. Niblett;L. Claflin
影响因子：
1.5
作者：
Jian-xiang Wu;Cui Yu;Cuiyun Yang;Xue-ping Zhou
通讯作者：
Jian-xiang Wu;Cui Yu;Cuiyun Yang;Xue-ping Zhou
影响因子：
13.8
作者：
Jian-xiang Wu;Lian Yu;Long Li;Jinqiang Hu;Jiyong Zhou;Xue-ping Zhou
通讯作者：
Jian-xiang Wu;Lian Yu;Long Li;Jinqiang Hu;Jiyong Zhou;Xue-ping Zhou
影响因子：
0.6
作者：
F. Morales;J. Arroyave;J. Castillo;C. León
通讯作者：
F. Morales;J. Arroyave;J. Castillo;C. León