Preparation of Monoclonal Antibody Against <i>Fusarium solani</i> and Development of Its Colloidal Gold Immunochromatographic Strip

doi:10.13523/j.cb.2202053

China Biotechnology

2022, Vol. 42

Issue (7): 54-61 DOI: 10.13523/j.cb.2202053

Preparation of Monoclonal Antibody Against Fusarium solani and Development of Its Colloidal Gold Immunochromatographic Strip

Kun WANG,Fu-yun ZHAO,Yun-fei XU,Xiao-feng YUAN,Wei-chun ZHAO^**(

)

School of Life Sciences, Zhejiang Chinese Medical University, Hangzhou 310053, China

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Abstract

Objective: The cell lines secreting specific monoclonal antibodies (McAbs) were prepared by using Fusarium solani, one of the pathogenic fungi causing root rot of Fritillaria thunbergii, and the colloidal gold immunochromatographic test strip based on McAbs was developed to provide scientific basis for detecting root rot of F. thunbergii. Methods: Hybridoma technology was used to obtain cell lines that could secrete specific McAbs against F. solani using the whole protein extract of F. solani as the antigen. The specificity, titer, sensitivity and binding protein of McAbs were detected by indirect ELISA and Western blot. Colloidal gold particles were prepared by trisodium citrate reduction method and McAbs were labeled to prepare colloidal gold immunochromatographic strip. Results: Three cell lines secreting specific McAbs against F. solani were obtained, which were named as FsA3, FsG6 and FsD4. The detection sensitivity of FsA3 was 24.41 ng / mL, and that of both FsG6 and FsD4 was 12.21 ng / mL. FsA3, FsG6 and FsD4 had strong reactions to F. solani, and had no cross-reaction to Alternaria tenuissima, A. alternata, Botrytis cinerea, F. equiseti, F. incarnatum, F. oxysporum, Phoma sp., and Phomopsis oblonga. The colloidal gold immunochromatographic strip based on FsG6 showed only a quality control line when detecting the tissue culture seedlings of F. thunbergii. When 100 ng F. solani antigen or the samples of F. thunbergii infested with root rot disease were detected, there were visible quality control lines and test lines. Conclusion: The specificity and sensitivity of the McAbs and test strip are sufficient to detect F. solani isolated from diseased strains of F. thunbergii, which provides the technical support for the rapid detection of root rot of F. thunbergii in the field.

Key words： Fritillaria thunbergia Root rot Fusarium solani Monoclonal antibody Gold immunochromatographic strip

Received: 28 February 2022 Published: 03 August 2022

ZTFLH:

Q816

Corresponding Authors: Wei-chun ZHAO E-mail: weichunzhao@zcmu.edu.cn

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Kun WANG,Fu-yun ZHAO,Yun-fei XU,Xiao-feng YUAN,Wei-chun ZHAO. Preparation of Monoclonal Antibody Against Fusarium solani and Development of Its Colloidal Gold Immunochromatographic Strip. China Biotechnology, 2022, 42(7): 54-61.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2202053 OR https://manu60.magtech.com.cn/biotech/Y2022/V42/I7/54

Fig.1 Detection of McAbs titer

Fig.2 McAbs sensitivity detection

Fig.3 Specific detection of McAbs

Fig.4 Type and subclass identification of McAbs

Fig.5 Western blot detection of McAbs binding protein

Fig.6 Effect of Fritillaria thunbergii Miq. on detection sensitivity of McAbs against F. solani

Fig.7 The effect of sample treatment on the detection sensitivity of McAb FsG6

Fig.8 Detection results of FsG6-based colloidal gold immunochromatographic strip for F. solani antigen a: Blank control b: Positive control

Fig.9 Field trial of colloidal gold immunochromatographic strip based on FsG6 a: Negative control b: Negative result c: Positive result


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