Antibody for detecting aspergillus

An antibody and Aspergillus technology, applied in the field of biochemistry, can solve problems such as limitations in popularization and application, and achieve the effects of simple operation, high specificity and low cost

Active Publication Date: 2008-03-26
ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, the popularization and

Method used

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  • Antibody for detecting aspergillus
  • Antibody for detecting aspergillus

Examples

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example 1

[0025] Example 1 Preparation and identification of monoclonal antibodies of the present invention

[0026] 1. Preparation of monoclonal antibody 9D47A1 and monoclonal antibody 7E11A1

[0027] 1) Preparation of Aspergillus fumigatus mycelium antigen:

[0028] The Aspergillus fumigatus strain was grown on a Sabouraud plate at 37°C for several days to form a single colony. Harvest a typical single colony, add it to a 500ml Erlenmeyer flask containing 50ml RPMI-1640, shake the bacteria for 3-7 days at 37°C, filter and separate the mycelium and filtrate. The collected hyphae were resuspended in PBS, lysed by ultrasound, and the supernatant was collected after centrifugation and stored at -80°C for later use.

[0029] 2) Immunize mice

[0030] Take 4-6 weeks old female BALB / c mice, use Freund’s complete adjuvant and equal volume of Aspergillus fumigatus mycelium antigen to mix and emulsify for the first time, subcutaneously inject 100μg / mouse, and then take Freund every 10 days. The inc...

example 2

[0063] Example 2 Double antibody sandwich ELISA kit for detecting Aspergillus of the present invention

[0064] 1. The double antibody sandwich ELISA kit for detecting Aspergillus consists of the following reagents:

[0065] Microwell reaction plate coated with monoclonal antibody 9D47A1

[0066] Sample treatment solution: 4% ethylenediaminetetraacetic acid (PH7.0), that is, weigh 40g of ethylenediaminetetraacetic acid, dissolve it in 1L distilled water, adjust the pH to 7.0 with NaOH, and store at 4℃;

[0067] Enzyme conjugate: horseradish peroxidase-labeled monoclonal antibody 7E11A1;

[0068] Concentrated lotion: 20×PBS containing 2% Tween-20, that is, 1L solution contains 4.56g NaH 2 PO 4 , 58.02gNa 2 HPO 4 .12H 2 O, 175.3g NaCl, 15 lbs. After autoclaving for 20 minutes, add 20ml Tween-20 and stir evenly, and dilute it by 20 times when used;

[0069] Positive control: Aspergillus fumigatus culture filtrate diluted 1:1000,

[0070] Negative control: 10mM pH7.4 PBS containing 0....

example 3

[0094] Example 3 Colloidal gold rapid immunochromatographic test paper for detecting Aspergillus

[0095] 1. The composition of the colloidal gold rapid immunochromatographic test paper for detecting Aspergillus according to the present invention is shown in Fig. 2. The test paper is composed of sample pad 1, polyester cellulose pad 2, nitrocellulose membrane 3 and absorbent pad 4 sequentially and partially overlapped. Composition, wherein a layer of colloidal gold-mAb 9D47A1 composite is sprayed on the polyester cellulose membrane 2 with a particle size of 18-20nm and is dark red; the nitrocellulose membrane 3 is distributed with a detection zone 5 and a quality control zone 6 The detection zone 5 is formed by spraying monoclonal antibody 7E11A1 on the nitrocellulose membrane 3, and the quality control zone 6 is formed by spraying goat anti-mouse IgG1 antibody on the nitrocellulose membrane 3.

[0096] The preparation method of the test paper of the present invention is: spraying...

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Abstract

This invention provides the capture antibody and the detection antibody to detect the aspergillus, the two antibody is the single clone antibody of the cross oncocyte CCTCC NO.C200731 and CCTCC NO.C200730, they can unites the glucoprotein whose molecular weight is 25-100kDa in the cell wall of the aspergillus, the reaction with the Penicillium marneffei, white oidiomycosis, smoothness oidiomycosis, tropic oidiomycosis, close smoothness oidiomycosis, Candida krusei are all negative. The antibody in this invention can make into the immunity reagent to detect the aspergillus in common use.

Description

Technical field [0001] The present invention relates to the field of biochemistry, in particular to a new antibody for detecting Aspergillus. Background technique [0002] Aspergillus is the most common saprophytic fungus in nature. There are more than 100 species, of which at least 10 species of Aspergillus are conditional pathogens, which often pollute food, the environment, and can infect immunosuppressed people, seriously threatening human health. Invasive Aspergillosis (IA) caused by Aspergillus has a high mortality rate of 95% due to the lack of effective early diagnosis and treatment methods. IA is mainly caused by Aspergillus fumigatus and Aspergillus flavus, and a small part is caused by Aspergillus terreus, Aspergillus niger, Aspergillus nidulans. Among them, Aspergillus fumigatus is the main pathogenic bacteria, accounting for 80% to 90% of human infections caused by Aspergillus fungi. . [0003] In terms of food testing, the current domestic and foreign methods for de...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/577G01N33/543C12N5/12
Inventor 车小燕郝卫潘玉先王艳芳
Owner ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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