Visual and Super-Sensitive Detection of Maize Chlorotic Mottle Virus by Dot-ELISA and Au Nanoparticle-Based Immunochromatographic Test Strip

被引:4
|
作者
Zhang, Cui [1 ]
Guo, Mengmeng [1 ]
Dong, Jinxi [1 ,2 ]
Liu, Li [3 ]
Zhou, Xueping [1 ,4 ]
Wu, Jianxiang [1 ,2 ]
机构
[1] Zhejiang Univ, Inst Biotechnol, State Key Lab Rice Biol, Key Lab Biol Crop Pathogens & Insects Zhejiang Pro, Hangzhou 310058, Peoples R China
[2] Zhejiang Univ, Hainan Inst, Sanya 572025, Peoples R China
[3] Zhejiang Econ & Trade Polytech, Dept Appl Engn, Hangzhou 310018, Peoples R China
[4] Chinese Acad Agr Sci, State Key Lab Biol Plant Dis & Insect Pests, Inst Plant Protect, Beijing 100193, Peoples R China
来源
VIRUSES-BASEL | 2023年 / 15卷 / 07期
关键词
maize chlorotic mottle virus; monoclonal antibody; Dot-ELISA; immunochromatographic test strip; RT-PCR; MEDIATED ISOTHERMAL AMPLIFICATION; SEROLOGICAL METHODS; POTATO-VIRUS; LETHAL NECROSIS; RAPID DETECTION; PLANTS; ASSAYS;
D O I
10.3390/v15071607
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Maize chlorotic mottle virus (MCMV) is the only species in the Mahromovirus genus and is often co-infected with one or several viruses of the Potyvirus genus, posing a great threat to the global maize industry. Effective viral integrated management measures are dependent on the timely and proper detection of the causal agent of the disease. In this work, six super-sensitive and specific monoclonal antibodies (mAbs) against MCMV were first prepared using purified MCMV virions as the immunogen. Then, the Dot enzyme-linked immunosorbent assay (Dot-ELISA) was established based on the obtained mAbs, and it can detect MCMV in infected maize leaf crude extracts diluted up to 1:10,240-fold (w/v, g/mL). Furthermore, a rapid and user-friendly Au nanoparticle-based immunochromatographic test strip (AuNP-ICTS) based on paired mAbs 7B12 and 17C4 was created for monitoring MCMV in point-of-care tests, and it can detect the virus in a 25,600-fold dilution (w/v, g/mL) of MCMV-infected maize leaf crude extracts. The whole test process for ICTS was completed in 10 min. Compared with conventional reverse transcription-polymerase chain reaction (RT-PCR), the detection endpoint of both serological methods is higher than that of RT-PCR, especially the Dot-ELISA, which is 12.1 times more sensitive than that of RT-PCR. In addition, the detection results of 20 blinded maize samples by the two serological assays were consistent with those of RT-PCR. Therefore, the newly created Dot-ELISA and AuNP-ICTS exhibit favorable application potential for the detection of MCMV in plant samples.
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页数:12
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