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Method for detecting circulating antigen for angiostrongylus cantonensis and enzyme-linked immunosorbent assay kit thereof

An enzyme-linked immunosorbent reagent, angiostrongylus technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of long detection time, cross-reaction, poor repeatability, etc., and achieve simple operation, strong specificity, and high sensitivity Effect

Inactive Publication Date: 2010-02-10
SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the traditional EIISA method has not been able to solve the problems of low sensitivity, relatively poor repeatability, easy cross-reaction with other parasitic diseases, and long detection time.

Method used

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  • Method for detecting circulating antigen for angiostrongylus cantonensis and enzyme-linked immunosorbent assay kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: A double-antibody sandwich method for detecting circulating antigens of Angiostrongylus cantonensis:

[0054] Referring to the figure, a double-antibody sandwich method for detecting circulating antigens of Angiostrongylus cantonensis comprises the following steps: (1) selecting two anti-Angiostrongylus cantonensis monoclonal antibodies against different determinants on the adult antigens of Angiostrongylus cantonensis, wherein the anti-Angiostrongylus cantonensis monoclonal antibody Angiostrongylus cantonensis monoclonal antibody A is used to coat the solid phase carrier, and anti-Angiostrongylus cantonensis monoclonal antibody B is used to prepare enzyme conjugates; (2) Drop onto the solid phase carrier (1) coated with anti-Angiostrongylus cantonensis monoclonal antibody A, add enzyme-labeled anti-Angiostrongylus cantonensis monoclonal antibody B (3) at the same time or after incubation, and incubate Finally, the positive specimen forms a sandwich complex o...

Embodiment 2

[0084] Example 2: Preparation and detection method of an enzyme-linked immunosorbent assay kit for detecting circulating antigens of Angiostrongylus cantonensis.

[0085] (1) An enzyme-linked immunosorbent assay kit for detecting circulating antigens of Angiostrongylus cantonensis, comprising:

[0086] ① ELISA plate coated with the original coating: Dilute the monoclonal antibody against the protein secretion of Angiostrongylus cantonensis adult to 2 μg with coating buffer (pH 9.6, 0.05 mol / L carbonate buffer) / ml, add 100μl to each well of a 96-well detachable polystyrene ELISA plate, incubate at 37°C for 2h, pour off the coating solution, wash 2-3 times with 19-fold diluted concentrated washing solution, each time for 30s, and pat dry , then add 200 μl of blocking solution (sample diluent), incubate at 37°C for 1 hour, pour off the liquid in the well, dry and store in a vacuum seal.

[0087] ②Enzyme marker: as described in Example 1, it is HRP marker;

[0088] ③Concentrate...

Embodiment 3

[0098] Embodiment three: detection of kit specificity, sensitivity, precision, stability:

[0099] ①Specific detection: 68 serum samples of SD rats infected with Angiostrongylus cantonensis, 50 serum samples of normal SD rats, 18 serum samples of patients with Angiostrongylus cantonensis, 80 samples of normal human serum, and 2 samples of adult Angiostrongylus cantonensis were selected. Antigen dilution, 2 parts of Angiostrongylus cantonensis protein secretion dilution, 2 parts of Schistosoma japonicum antigen dilution, 2 parts of human roundworm antigen dilution, 2 parts of liver fluke antigen dilution, 24 parts of paragonimiasis patient serum, 18 parts Serum from schistosomiasis patients, 38 serums from liver fluke patients, 27 serums from ascariasis patients, 12 serums from cysticercosis patients, 7 serums from echinococcosis patients, 5 serums from patients with filariasis, prepared in Example 2 The enzyme-linked immunosorbent assay kit for detecting the circulating antige...

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Abstract

The invention relates to a double antibody sandwich method for detecting circulating antigen for angiostrongylus cantonensis and an enzyme-linked immunosorbent assay kit thereof, which belong to the technical field of enzyme immunoassay. The enzyme-linked immunosorbent assay kit for detecting the antigen for the angiostrongylus cantonensis comprises an ELIAS plate enveloped by an envelope antigen,and an enzyme marker, wherein the envelope antigen and the enzyme marker are both monoclonal antibodies for an angiostrongylus cantonensis protein secretion. The detection specificity of the kit reaches 100 percent; the precision (coefficient of variation C.V) is 8.54 percent; and the kit has simple and convenient operation, is rapid, can finish a one-step detection method within 40 minutes, andcan realize the multi-sample diagnosis and curative effect evaluation of angiostrongylus cantonensis.

Description

technical field [0001] The invention relates to a detection method for circulating antigen of Angiostrongylus cantonensis and an enzyme-linked immunosorbent reagent kit thereof, which are mainly suitable for rapid diagnosis and evaluation of curative effect of Angiostrongylus cantonensis. Background technique [0002] Angiostrongylus cantonensis was discovered and named by the Chinese scholar Chen Xintao (1933) in the lungs of the Guangzhou house mouse. It is mainly distributed in the Pacific Ocean, some islands in the Indian Ocean and Southeast Asian countries. Human infection with Angiostrongylus cantonensis is mainly caused by eating raw or half-cooked snails, fish, and shrimps containing third-stage larvae, as well as vegetables, fruits and drinking water contaminated by this larvae. The first human case of angiostrongyliasis cantonensis was discovered in Taiwan by Nomura and Lin. Since He Jingzhi reported the first case of Angiostrongylus cantonensis in mainland China...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N33/558
Inventor 陈木新朱兴全张仁利
Owner SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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