Cystic echinococcosis cyst fluid degreasing antigen for detecting alveolar echinococcosis antibody, and preparation method thereof

Abstract

The invention relates to the technical field of human and animal alveolar echinococcosis (AE) antibody detections, specifically to cystic echinococcosis (CE) cyst fluid degreasing antigen for detecting AE antibody, and a preparation method thereof. According to the present invention, the CE cyst fluid degreasing antigen is used for detecting AE antibody by using an enzyme-linked immunosorbent test double antigen sandwich method so as to effectively increase quality of the detection reagent, and provide characteristics of strong specificity, high sensitivity, stable performance and wide uses; the CE cyst fluid degreasing antigen can be used for detecting human and animal AE antibodies so as to conveniently compare antibody contents of different animal varieties; and according to the immunological reaction principle, the CE cyst fluid degreasing antigen further can be used for preparing colloidal gold immunochromatography reagents and other rapid detection reagents for detecting AE antibodies so as to create favorable conditions for echinococcosis identification diagnosis, prevalence surveys and epidemic situation surveillance works.

Description

technical field

[0001] The invention relates to the technical field of human and animal alveolar echinococcosis antibody detection, and relates to a cyst fluid deesterified antigen of cystic echinococcosis suitable for detecting alveolar echinococcosis antibody and a preparation method thereof. Background technique

[0002] Alveolar echinococcosis (AE) is a zoonotic disease caused by the larval parasitism of Echinococcus multilocularis (Em), with no obvious symptoms in the early stage, and most of the lesions have spread when discovered, and the lesions are progressive Infiltrating growth, conventional diagnostic methods such as ultrasound and CT examination are not easy to distinguish from tumor lesions, and serological testing is helpful for the diagnosis of AE. The detection of echinococcosis antibody is an important indicator for the diagnosis of echinococcosis, and the antigen is a key reagent that affects the specificity and sensitivity of detection. Methods such as s...

Examples

Embodiment 1

[0018] Example 1, a cyst fluid deesterified antigen of cystic echinococcosis suitable for detecting alveolar echinococcosis antibody was obtained according to the following steps:

[0019] The first step is to collect the cyst fluid of sheep cystic echinococcosis, add solid ammonium sulfate to the cyst fluid to dissolve, and make the concentration of ammonium sulfate in the cyst fluid reach 30% of the saturated concentration, and then use sodium hydroxide or ammonium hydroxide to dissolve the cyst fluid. Adjust to pH 7.2, then add chloroform or petroleum ether in an amount of 30% to 50% of the liquid volume after pH adjustment, shake vigorously to form a suspension, and let the suspension stand until the chloroform or petroleum ether and the encapsulation liquid are separated. Vigorously oscillate again and let stand to separate layers for 2 times, then centrifuge at 4000g for 20min, discard the precipitate and chloroform or petroleum ether, and collect the treated encapsulatio...

Embodiment 2

[0024] Example 2, the application of the cystic echinococcosis cyst fluid deesterified antigen suitable for detecting alveolar echinococcosis antibody obtained in the above-mentioned embodiment 1 in the detection of alveolar echinococcosis antibody; its application is as follows:

[0025] 1. ELISA sandwich assay kit for detecting alveolar echinococcosis antibody and preparation method thereof

[0026] ELISA sandwich assay kit for detection of alveolar echinococcosis antibody, including 8 tubes x 12 strips of cystic echinococcosis cyst fluid deesterified antigen-coated plate, horseradish peroxidase labeled cystic echinococcosis Echinococcosis cyst fluid deesterified antigen 10ml, specimen diluent 10ml, 20 times concentrated washing solution 50ml, alveolar echinococcosis antibody positive control solution 3ml, alveolar echinococcosis antibody negative control solution 3ml, cystic echinococcosis for inhibition Disease capsule fluid deesterified antigen 6ml, chromogenic solution A...

technical field

[0001] The invention relates to the technical field of human and animal alveolar echinococcosis antibody detection, and relates to a cyst fluid deesterified antigen of cystic echinococcosis suitable for detecting alveolar echinococcosis antibody and a preparation method thereof. Background technique

[0002] Alveolar echinococcosis (AE) is a zoonotic disease caused by the larval parasitism of Echinococcus multilocularis (Em), with no obvious symptoms in the early stage, and most of the lesions have spread when discovered, and the lesions are progressive Infiltrating growth, conventional diagnostic methods such as ultrasound and CT examination are not easy to distinguish from tumor lesions, and serological testing is helpful for the diagnosis of AE. The detection of echinococcosis antibody is an important indicator for the diagnosis of echinococcosis, and the antigen is a key reagent that affects the specificity and sensitivity of detection. Methods such as s...

Examples

Embodiment 1

[0018] Example 1, a cyst fluid deesterified antigen of cystic echinococcosis suitable for detecting alveolar echinococcosis antibody was obtained according to the following steps:

[0019] The first step is to collect the cyst fluid of sheep cystic echinococcosis, add solid ammonium sulfate to the cyst fluid to dissolve, and make the concentration of ammonium sulfate in the cyst fluid reach 30% of the saturated concentration, and then use sodium hydroxide or ammonium hydroxide to dissolve the cyst fluid. Adjust to pH 7.2, then add chloroform or petroleum ether in an amount of 30% to 50% of the liquid volume after pH adjustment, shake vigorously to form a suspension, and let the suspension stand until the chloroform or petroleum ether and the encapsulation liquid are separated. Vigorously oscillate again and let stand to separate layers for 2 times, then centrifuge at 4000g for 20min, discard the precipitate and chloroform or petroleum ether, and collect the treated encapsulatio...

Embodiment 2

[0024] Example 2, the application of the cystic echinococcosis cyst fluid deesterified antigen suitable for detecting alveolar echinococcosis antibody obtained in the above-mentioned embodiment 1 in the detection of alveolar echinococcosis antibody; its application is as follows:

[0025] 1. ELISA sandwich assay kit for detecting alveolar echinococcosis antibody and preparation method thereof

[0026] ELISA sandwich assay kit for detection of alveolar echinococcosis antibody, including 8 tubes x 12 strips of cystic echinococcosis cyst fluid deesterified antigen-coated plate, horseradish peroxidase labeled cystic echinococcosis Echinococcosis cyst fluid deesterified antigen 10ml, specimen diluent 10ml, 20 times concentrated washing solution 50ml, alveolar echinococcosis antibody positive control solution 3ml, alveolar echinococcosis antibody negative control solution 3ml, cystic echinococcosis for inhibition Disease capsule fluid deesterified antigen 6ml, chromogenic solution A...