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

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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.

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Authors and Affiliations

  1. State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou, 310058, China

    Jian-xiang Wu, Qiang Wang, Huan Liu, Ya-juan Qian, Yan Xie & Xue-ping Zhou

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  1. Jian-xiang Wu
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  2. Qiang Wang
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  3. Huan Liu
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  4. Ya-juan Qian
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  5. Yan Xie
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  6. Xue-ping Zhou
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Corresponding authors

Correspondence to Yan Xie or Xue-ping Zhou.

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The two authors contributed equally to this work

Project supported by the National Natural Science Foundation of China (No. 31272015), the Ministry of Education of China (No. 313052), and the Zhejiang Provincial Natural Science Foundation of China (No. Z3090039)

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Wu, Jx., Wang, Q., Liu, H. et al. Monoclonal antibody-based serological methods for maize chlorotic mottle virus detection in China. J. Zhejiang Univ. Sci. B 14, 555–562 (2013). https://doi.org/10.1631/jzus.B1200275

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  • DOI: https://doi.org/10.1631/jzus.B1200275

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