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Color microsphere immunochromatography test paper for rapidly diagnosing entamoeba histolytica antigen and preparation method thereof

A technology for immunochromatographic detection and amoeba, which is applied in the field of immunodiagnosis, can solve the problems of being unsuitable for primary detection, long detection time, and lack of specificity, and achieves reduced investment and detection costs, short detection time, and high specificity strong effect

Pending Publication Date: 2020-11-06
HANGZHOU ALLTEST BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The direct smear method is easy to operate and low in cost, but the detection rate is low; the three methods of cyst concentration method, biopsy, and nucleic acid diagnosis are cumbersome to operate; indirect hemagglutination (IHA) and enzyme-linked immunosorbent assay ( Although ELISA) has high sensitivity, it lacks specificity and is easy to be misdiagnosed; these methods are suitable for laboratory research, and the detection time is long, so they are not suitable for basic level detection. Therefore, there is an urgent need to develop a simpler and faster detection method

Method used

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  • Color microsphere immunochromatography test paper for rapidly diagnosing entamoeba histolytica antigen and preparation method thereof
  • Color microsphere immunochromatography test paper for rapidly diagnosing entamoeba histolytica antigen and preparation method thereof
  • Color microsphere immunochromatography test paper for rapidly diagnosing entamoeba histolytica antigen and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The preparation method of the color microsphere immunochromatography test paper for rapid diagnosis of Entamoeba histolytica specifically comprises the following steps:

[0044] 1) Preparation of test line T and quality control line C: Dilute mouse anti-Entamoebic histolytica antibody and streptavidin-rabbit IgG conjugate solution respectively and spray them on nitric acid at a rate of 1.0ul / cm The cellulose membrane was used as the test line T and the quality control line C respectively, and dried at 36°C to obtain the test line and the quality control line; the test line T was coated with mouse anti-Entamoeba histolytica antibody The concentration is 0.1 mg / ml; the concentration of streptavidin-rabbit IgG conjugate coated with the quality control line C is 1.0 mg / ml;

[0045] 2) Preparation of the marker pad: when the diluted colored microsphere-antibody conjugate is at a concentration of 0.1%, it is treated on a polyester cellulose membrane at a rate of 2.0 ul / cm, an...

Embodiment 2

[0049] The difference from Example 1 is that the preparation method of the color microsphere immunochromatography test paper for rapid diagnosis of Entamoeba histolytica specifically includes the following steps:

[0050] 1) Preparation of test line T and quality control line C: Dilute mouse anti-Entamoebic histolytica antibody and streptavidin-rabbit IgG conjugate solution respectively and spray them on nitric acid at a rate of 1.05ul / cm The cellulose membrane was used as the test line T and the quality control line C respectively, and dried at 36.5°C to obtain the test line and the quality control line; the test line T was coated with mouse anti-Entamoeba histolytica antibody The concentration is 0.15 mg / ml; the concentration of streptavidin-rabbit IgG conjugate coated with the quality control line C is 1.2 mg / ml;

[0051] 2) Preparation of the marker pad: when the diluted colored microsphere-antibody conjugate was at a concentration of 0.15%, it was treated on a polyester c...

Embodiment 3

[0055] The difference from Example 1 is that the preparation method of the color microsphere immunochromatography test paper for rapid diagnosis of Entamoeba histolytica specifically includes the following steps:

[0056] 1) Preparation of test line T and quality control line C: Dilute mouse anti-Entamoebic histolytica antibody and streptavidin-rabbit IgG conjugate solution respectively and spray them on nitric acid at a rate of 1.1ul / cm The cellulose membrane was used as the test line T and the quality control line C respectively, and dried at 37°C to obtain the test line and the quality control line; the test line T was coated with mouse anti-Entamoeba histolytica antibody The concentration is 0.2 mg / ml; the concentration of streptavidin-rabbit IgG conjugate coated with the quality control line C is 1.5 mg / ml;

[0057] 2) Preparation of the marker pad: Dilute the colored microsphere-antibody conjugate to a concentration of 0.2%, process it on the polyester cellulose membrane...

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Abstract

The invention relates to the technical field of immunodiagnosis. In view of the blanks in the prior art, the invention discloses color microsphere immunochromatography test paper for rapidly diagnosing entamoeba histolytica antigen and a preparation method thereof. The method specifically comprises the following steps of (1) respectively spraying a mouse anti entamoeba histolytica monoclonal antibody and a streptavidin-rabbit IgG conjugate solution onto a nitrocellulose membrane to respectively serve as a detection line T and a quality control line C, 2) diluting the mouse anti entamoeba histolytica monoclonal antibody-color microsphere conjugate to a concentration of 0.1-0.3%, spraying the diluted conjugate on a polyester cellulose membrane at a speed of 2.0-3.0 mul / cm, and drying to obtain a marking pad, and 3) preparing the detection test paper. The method has the characteristics of simplicity, convenience and quickness in operation, intuitive and accurate result display, strong specificity, simple preparation process, reduction of investment and detection cost, wide application range and the like, and can also be widely applied to basic-layer sample detection.

Description

technical field [0001] The invention relates to the technical field of immunodiagnosis, in particular to a color microsphere immunochromatographic detection test paper for rapidly diagnosing Entamoeba histolytica antigen and a preparation method thereof. Background technique [0002] Entamoeba histolytica, also known as dysentery amoeba, mainly parasitizes the colon, causing amoebic dysentery and parenteral amoebiasis. About 400 to 500 million people in the world are infected, and about 40000 to 100000 people die every year. The global distribution of the disease is more common in subtropical tropical fish, and its harm is second only to malaria. [0003] The pathogen of amoebiasis is Entamoeba histolytica. Amoeba protozoa exists in human body in two forms: trophozoite and cyst. After infection with amoeba, severe invasive amoebiasis can appear symptom. It can also be asymptomatic with worms. [0004] Histolytic amebiasis is mainly caused by amoeba protozoa polluted wate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/558G01N33/543
CPCG01N33/56905G01N33/558G01N33/54313
Inventor 陈金树陆维克张芳高飞
Owner HANGZHOU ALLTEST BIOTECH
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