阅读说明：本技术 识别茄病镰刀菌的单克隆抗体及其杂交瘤细胞株FsG6 (Monoclonal antibody for identifying fusarium solani and hybridoma cell strain FsG6 thereof ) 是由 赵伟春 祁依佳 黄唯一 葛佳敏 徐云飞 于 2019-11-27 设计创作，主要内容包括：本发明公开了一种识别茄病镰刀菌Fusarium solani的单克隆抗体及其杂交瘤细胞株,所述识别茄病镰刀菌的单克隆抗体,由保藏编号为：CCTCC No：C2019225的杂交瘤细胞株分泌产生；所述杂交瘤细胞株命名为杂交瘤细胞株FsG6,于2019年10月17日保藏于中国典型培养物保藏中心(CCTCC),保藏编号为：CCTCC No：C2019225。该单克隆抗体被用于由茄病镰刀菌侵染引发的动植物病害的鉴定、动态监测以及茄病镰刀菌的生物学研究,通过BaL b/c小鼠腹腔注射该细胞株,即可获得大量的该单克隆抗体,与多克隆抗体相比,具有纯度高,专一性强,重复性好等优点。(The invention discloses a monoclonal antibody for identifying Fusarium solani and a hybridoma cell strain thereof, wherein the monoclonal antibody comprises the following preservation numbers: CCTCC No: c2019225 hybridoma cell strain secretion; the hybridoma cell strain is named as hybridoma cell strain FsG6, is preserved in China Center for Type Culture Collection (CCTCC) in 2019, 10 and 17 months, and has the preservation number as follows: CCTCC No: C2019225. the monoclonal antibody is used for identification and dynamic monitoring of animal and plant diseases caused by fusarium solani infection and biological research of fusarium solani, and a large amount of monoclonal antibody can be obtained by injecting the cell strain into the abdominal cavity of a Balb/c mouse.)

1. A monoclonal antibody for identifying fusarium solani is disclosed by the preservation number: CCTCC No: c2019225 hybridoma cell strain secretion.

2. A hybridoma cell strain is named as hybridoma cell strain FsG6, is preserved in China Center for Type Culture Collection (CCTCC) in 2019, 10 months and 17 days, and has the preservation number as follows: CCTCC No: C2019225.

Technical Field

The invention belongs to the field of animals, and particularly relates to a monoclonal antibody for identifying fusarium solani and a hybridoma cell strain FsG6 thereof.

Background

Fusarium solani, a fungus of the genus Fusarium of the class Deuteromycotina, is one of the pathogenic fungi of fritillary root rot (also known as dry rot). The pathogenic bacteria are inhabitant bacteria. Pathogenic fungi spread through soil transfer as mycelia, spores and chlamydospores and invade the host mainly through wounds of plants. Long distance transmission is performed through diseased seed stems. The fritillary root rot is one of the important diseases of fritillary, can damage fritillary in the field production period and the storage period, and the damaged bulbs are cellular, thus seriously affecting the yield and the quality of the fritillary. Because the root rot disease is mainly underground bulbs, the field symptoms are difficult to find in early stage, and fusarium fungi are various in variety, and the bacterial colonies, hyphae and spores of closely related species are similar in shape and difficult to distinguish by means of morphological characteristics, the early diagnosis of the root rot and the classification and identification of pathogenic bacteria always restrict the scientific and effective prevention and control of the root rot of fritillaria. The monoclonal antibody (monoclonal antibody) combined with enzyme-linked immunosorbent assay (ELISA) for fusarium solani disclosed by the invention has the characteristics of high sensitivity and strong specificity, and is suitable for detecting a large number of plant, animal and microorganism samples, so that a foundation is laid for identification and biological research of fusarium solani by using the antibody and dynamic monitoring of diseases infected by fusarium solani, such as fritillaria root rot and the like.

Disclosure of Invention

The invention provides a monoclonal antibody for identifying fusarium solani, which is prepared from the following components in parts by weight: CCTCC No: c2019225 hybridoma cell strain secretion.

The invention also provides a hybridoma cell strain which is named as hybridoma cell strain FsG6 and is preserved in China Center for Type Culture Collection (CCTCC) in 2019, 10 months and 17 days, wherein the preservation numbers are as follows: CCTCC No: C2019225.

the invention has the following beneficial effects:

(1) the monoclonal antibody has strong specificity: the monoclonal antibody has strong reaction with fusarium solani antigens, does not react with the antigens of alternaria, alternaria tenuissima, fusarium oxysporum, fusarium equiseti, fusarium semitectum, botrytis cinerea, phoma and phomopsis, and can be used for detecting the fusarium solani and the root rot of fritillaria thunbergii caused by the fusarium solani.

(2) The detection sensitivity of the monoclonal antibody is high: the indirect ELISA detection result shows that the detection sensitivity of the monoclonal antibody to the antigen prepared from fusarium solani hyphae and spores is 12.21ng/mL (namely 1.221ng per hole), and the monoclonal antibody has good development and application prospects.

(3) The monoclonal antibody is IgG2 a.

(4) The monoclonal antibody-bound target protein is single: the monoclonal antibody only binds to an antigen protein of about 43kDa of fusarium solani.

(5) The detection sensitivity of the thunberg fritillary bulb to the monoclonal antibody has no influence: the monoclonal antibody has no cross reaction to the thunberg fritillary bulb protein extracting solution; after the thunberg fritillary bulb protein extracting solution is added into the antigen prepared from fusarium solani hyphae and spores, the influence of the thunberg fritillary bulb extracting solution on the detection sensitivity of the monoclonal antibody is small, the detection sensitivity reaches 12.21ng/mL (namely 1.221ng per hole), and the method has a good development and application prospect.

Drawings

Potency assay of FIG. 1FsG 6;

FIG. 2FsG6 reaction with different fungal antigens;

the detection sensitivity assay of FIG. 3FsG 6;

type and subclass identification of FIG. 4FsG 6;

FIG. 5FsG6 detection of bound target protein;

FIG. 6 shows the effect of Bulbus Fritillariae Thunbergii extract on FsG6 detection sensitivity.

Detailed Description

The invention is further explained below with reference to the figures and the examples.