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Monoclonal antibody for resisting acidovorax citrulli and application thereof

A technology of monoclonal antibody against melon fruit spot bacteria, applied in the direction of anti-bacterial immunoglobulin, microbial-based methods, biochemical equipment and methods, etc., to achieve good broad-spectrum, high titer, and avoid cross-reaction effects

Inactive Publication Date: 2014-06-25
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] For this bacterial disease, a more feasible control measure is to implement quarantine control and its expansion, which depends on the application of accurate, rapid and sensitive detection technology

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1, the acquisition of mouse hybridoma cell line and the preparation of the monoclonal antibody against melon fruit blight

[0054] 1. Animal immunity

[0055] 1. Bacterial culture

[0056]Pick a single colony of Acidovorax citrulli ATCC29625 on the KB plate medium, streak it on the KB plate medium, incubate at 26°C for 48 hours, wash the colony with normal saline into a 2ml centrifuge tube, wash with normal saline 2 times, adjust the concentration to 10 8 CFU / mL, used in mouse immunoassay.

[0057] 2. Immunization of mice

[0058] The first immunization: Take an equal volume of Freund's complete adjuvant and the bacterial suspension of P. melonifolia obtained in step 1, and inject Balb / c mice intraperitoneally after emulsification is complete. The dosage for each mouse is 10 8 CFU Melon fruit blotch.

[0059] The second immunization: 2 weeks after the first immunization, the second immunization was carried out, and an equal volume of Freund's incomplete ...

Embodiment 2

[0091] The detection of the monoclonal antibody of the anti-cucurbit fruit spot bacterium that embodiment 2, embodiment 1 obtain

[0092] One, the specificity detection of the monoclonal antibody of the anti-cucurbit fruit spot bacterium that embodiment 1 obtains

[0093] 1. Tested strains

[0094] Acidovorax citrulli ATCC 29625 and the control bacteria in Table 1.

[0095] Table 1 The control bacteria used in the specific detection of monoclonal antibodies against Melon fruit spot bacteria

[0096] serial number

scientific name

Chinese name

1

Acidovorax cattleyae

Oat Acidovorax subsp. Cartleylan ATCC10200

2

Acidovorax konjaci

Acidovorax subspecies Amorphophallus ATCC33996

3

Pseudomonas marginalis pv. marginalis

Bacterial leaf blight NCPPB247

[0097] 4

Pseudomonas syringae

Bacterial keratosis sp. ATCC9004

5

Pseudomonas viridiflava

Bacillus wilt ATCC13222

6

Pseudo...

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Abstract

The invention discloses a monoclonal antibody for resisting acidovorax citrulli and application thereof. The monoclonal antibody for resisting the acidovorax citrulli provided by the invention is secreted by a hybridoma cell strain 4C6 for secreting the monoclonal antibody of acidovorax citrulli; the mouse hybridoma cell strain is collected in China General Microbiological Culture Collection Center (CGMCC) with the collection number of CGMCC No.8960. The monoclonal antibody for resisting acidovorax citrulli provided by the invention is good in specificity and high in valence, reacts with different separators of the acidovorax citrulli, and has good broad-spectrum performance. The monoclonal antibody is applied to an acidovorax citrulliserological diagnosis reagent, good in specificity, and avoids cross reaction. The monoclonal antibody for resisting acidovorax citrulli developed by the invention can be used for developing immunological diagnostic reagents such as an ELISA diagnosis reagent, a colloidal gold immune test paper strip, an antibody chip, a biological sensor, and the like, of the acidovorax citrulli.

Description

technical field [0001] The invention relates to a monoclonal antibody against melon fruit spot pathogen and application thereof. Background technique [0002] Melon fruit spot disease is a devastating disease on watermelon and muskmelon, which is caused by Acidovorax citrulli and is an important domestic and foreign plant quarantine pest in my country. The disease is mainly transmitted by seed borne fungus. Both seedling and adult plants can be affected. Irregular brown lesions appeared along the midrib of melon seedlings infected with the disease, and some extended to the edge of the leaf, which was water-soaked from the back of the leaf. The melon seedlings with seeds infected with bacteria died 1 to 3 weeks after the onset. When the watermelon fruit is infected, several millimeters in size gray-green to dark green water-soaked spots appear on the upper surface of the fruit, and then rapidly expand into large irregular water-soaked spots, which turn brown or crack, causi...

Claims

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Application Information

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IPC IPC(8): C12N5/20C07K16/12G01N33/577G01N33/535C12R1/91
Inventor 李桂芬马洁田茜魏梅生赵文军
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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