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Gold mark reagent kit for detecting angiostrongyliasis cantonensis and preparing method thereof

An angiostrongylia and kit technology, which is applied in the field of gold standard kits and preparations for the detection of angiostrongyliasis in Guangzhou, can solve the problems such as being unsuitable for on-site sample analysis, confined in the laboratory, and complicated in the operation process. Practical value, no need for special equipment, good repeatability

Inactive Publication Date: 2009-04-15
SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are some deficiencies in the traditional EIISA method: (1) the sensitivity and specificity of detection need to be further improved, and the repeatability is not good enough; (2) special equipment such as microplate reader, incubator, etc. are needed to cooperate with it; ( 3) Operators need to undergo special training, the operation process is relatively complicated, and the detection time is relatively long; (4) The detection process is limited in the laboratory, which is not suitable for the analysis of on-site samples

Method used

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  • Gold mark reagent kit for detecting angiostrongyliasis cantonensis and preparing method thereof
  • Gold mark reagent kit for detecting angiostrongyliasis cantonensis and preparing method thereof
  • Gold mark reagent kit for detecting angiostrongyliasis cantonensis and preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: A gold-labeled rapid diagnostic kit for detecting Angiostrongylus cantonensis

[0040] refer to figure 1 , Guangzhou Angiostrongylus gold standard rapid diagnostic kit with Guangzhou Angiostrongylus gold standard detection plate (1)

[0041] refer to figure 2 , the Angiostrongylus guangzhou gold standard detection plate (1) is provided with a sample end absorbent pad (4), a nitrocellulose membrane detection layer (6) and a terminal absorbent pad (9) on a PVC backing plate (10). A gold-labeled anti-Angiostrongylus cantonensis monoclonal antibody pad (5) is provided between the sample end absorbent pad (4) and the nitrocellulose membrane detection layer (6), and a detection line is coated on the nitrocellulose membrane detection layer (6) (7) and quality control line (8). Wherein the sample end absorbent pad (4) and the terminal absorbent pad (9) are made of multi-layer filter paper, and the gold standard pad (5) is made of glass fiber soaked with gold stan...

Embodiment 2

[0048] Embodiment 2: The preparation method of Angiostrongylus cantonensis Gold Standard Rapid Diagnostic Kit mainly includes the following steps:

[0049] (1) Preparation of soluble antigen of Angiostrongylus cantonensis:

[0050] Isolate the third-stage larvae of Angiostrongylus cantonensis from the snails of Agate snails or apple snails, and artificially infect SD rats at an amount of 80-100 per snail. After 33-35 days, adults of Angiostrongylus cantonensis are isolated from the pulmonary artery of the rats, normal saline After rinsing, freeze-dry, grind into powder, degrease with cold acetone 2-3 times, freeze-thaw repeatedly after ultrasonic crushing, centrifuge at 14000rpm 4°C for 20min, the supernatant is the soluble antigen of Angiostrongylus cantonensis, and store it at -20°C until use.

[0051] (2) The preparation and purification method of anti-Angiostrongylus cantonensis monoclonal antibody is the same as the example

[0052] (3) Preparation of 40nm colloidal gold...

Embodiment 3

[0065] Example 3: Use of Angiostrongylus guangzhou Gold Standard Rapid Diagnostic Kit:

[0066] Take out the test kit, put it flat on the operating table, add 2 drops (about 100 μl) of human or rodent cerebrospinal fluid and serum to the sample hole, and the result can be judged within 5 to 15 minutes. Judgment of results: In the observation hole, both the test line (T) and the quality control line (C) appear purple-red lines (such as image 3 ), it is judged as positive, the higher the concentration of circulating antigen, the darker the color of the detection line (T); and in the observation hole, only the quality control line (C) appears a purple-red line (such as Figure 4 ), it is judged as negative; when neither the quality control line (C) nor the test line (T) in the observation hole shows a purple line or only the test line (T) shows a color line, it means that the kit is invalid.

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Abstract

The invention relates to a gold-labeled kit for detecting angiostrongylus cantonensis disease and a preparation method thereof. The method comprises the preparation of angiostrongylus cantonensis soluble antigen, the preparation and purification of monoclonal antibodies, the preparation of colloidal gold-labeled anti-angiostrongylus cantonensis monoclonal antibodies, and the preparation of an angiostrongylus cantonensis gold-labeled rapid diagnosis kit. The kit comprises an angiostrongylus cantonensis gold-labeled detection plate; the detection plate comprises a PVC liner plate, an absorbent pad, a gold-labeled pad which is coated by colloidal gold-labeled angiostrongylus cantonensis monoclonal antibodies, a detection line which coats angiostrongylus cantonensis monoclonal antibodies inside and a quality control line of goat anti-mouse IgG or IgM. The kit is simple and rapid in operation, requiring only 5 to 15min; the kit has high sensitivity and strong specificity, and the detection rate of the rat serum which is experimentally infected by angiostrongylus cantonensis and the detection rate of the angiostrongylus cantonensis patient serum are both 100 percent. The kit and the preparation method can play an important role in rapid diagnosis and efficacy assessment of angiostrongylus cantonensis.

Description

technical field [0001] The invention relates to a gold standard kit for detecting angiostrongylus cantonensis and a preparation method thereof, which can realize rapid diagnosis and curative effect assessment of angiostrongylus cantonensis. Background technique [0002] Angiostrongylus cantonensis (Angiostrongylus cantonensis) was discovered and named by Chinese scholar Chen Xintao in the lungs of house mice in Guangzhou in 1933. It is mainly distributed in the Pacific Ocean, some islands in the Indian Ocean and Southeast Asian countries. The intermediate hosts of Angiostrongylus cantonensis are mainly snails and slugs, and the final hosts are mainly rodents, especially rats. Adults parasitize in the pulmonary arteries, and people are mainly infected by eating raw or half-cooked snails, fish, shrimps containing the third-stage larvae, vegetables, fruits, and drinking water contaminated by the third-stage larvae, and the larvae invade Central nervous system can cause eosino...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/569G01N33/558
Inventor 张仁利朱兴全陈木新
Owner SHENZHEN CENT FOR DISEASE CONTROL & PREVENTION
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