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Colloidal gold method detection test strip and reagent kit for IgM and IgG antibodies of mycoplasma pneumoniae and preparation method of reagent kit

A technology of Mycoplasma pneumoniae and detection kit, which is applied to measurement devices, instruments, scientific instruments and other directions, can solve the problems of cumbersome operation process, insufficient detection convenience and quickness, etc., and achieves the effect of simplified operation process, simple detection and high sensitivity

Inactive Publication Date: 2013-02-13
JIANGSU KEYGEN BIOTECH CORP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the individual detection of IgM and IgG antibodies, and then a comprehensive analysis of the test results has no effect on the diagnosis of the disease, but the operation process is cumbersome and the detection is not convenient and quick

Method used

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  • Colloidal gold method detection test strip and reagent kit for IgM and IgG antibodies of mycoplasma pneumoniae and preparation method of reagent kit
  • Colloidal gold method detection test strip and reagent kit for IgM and IgG antibodies of mycoplasma pneumoniae and preparation method of reagent kit
  • Colloidal gold method detection test strip and reagent kit for IgM and IgG antibodies of mycoplasma pneumoniae and preparation method of reagent kit

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: The composition of Mycoplasma pneumoniae IgM, IgG antibody detection kit (colloidal gold method)

[0032] 1. Sample processing bottle: The sample processing bottle (purchased from Hangzhou Beiyu) is filled with 1mL sample processing solution (phosphate buffer solution, the main raw material is purchased from Sinopharm Group).

[0033] 2. Test strip: 60*5mm bottom plate (purchased from Shanghai Jieyi Company); 15*5mm thick fiber filter paper for absorbent pad (purchased from Shanghai Jieyi Company); 25*5mm nitrocellulose membrane (purchased from Millipore Company ) were sequentially coated with mouse anti-human IgG antibody (purchased from ABD Company), mouse anti-human IgM antibody (purchased from ABD Company), rabbit anti-MP polyclonal antibody (purchased from ABCAM Company); sample pad (purchased from Shanghai Jieyi Company ) was attached with a whole blood filtration membrane (purchased from Shanghai Jieyi Company); the gold standard pad (purchased from...

Embodiment 2

[0034] Embodiment 2: the preparation of kit

[0035] 1. Preparation of sample processing solution: Prepare sample processing solution: prepare phosphate buffer solution (pH7.0±0.2), dispense it into sample processing bottles, 1mL / bottle.

[0036] 2. Preparation of test strips:

[0037] 2.1. Dilute the mouse anti-human IgG antibody, mouse anti-human IgM antibody, and rabbit anti-MP polyclonal antibody to a coating concentration of 0.03-0.05 mg / mL, and draw the three coating solutions on the designated positions on the nitrocellulose membrane respectively. The coated nitrocellulose membrane was dried and stored at 37°C for 4 hours.

[0038]2.2. Preparation of gold standard pad: Colloidal gold was prepared by trisodium citrate reduction method, and recombinant MP antigen was slowly added to the gold sol at a ratio of 20 μg / mL to 40 μg / mL. After fully reacting, centrifuge at 8000 RPM for 30 minutes, discard Add the diluent to 1 / 3 of the original volume, then soak the gold standa...

Embodiment 3

[0043] Embodiment 3 detection method

[0044] 1. Test samples according to clinical routines. The test samples can be serum, whole blood, sputum, and throat swabs. If the sample cannot be detected within three days, it can be stored at 2-8°C, and the storage period should not exceed 7 days.

[0045] 2. Take the sputum and throat test into the sample processing bottle, shake the specimen well, and let it stand for 5 minutes. Serum and whole blood samples do not require this step.

[0046] 3. Take 100-200 ul of the processed sample or serum or whole blood sample and add dropwise to the sample loading window. If the whole blood is relatively viscous, add 1 to 2 drops of unused sample processing solution to the sample loading window.

[0047] 4. Interpret the result after 10-15 minutes:

[0048] 4.1. If the sample contains Mycoplasma pneumoniae IgG antibody, it will combine with the recombinant MP antigen-colloidal gold complex when passing through the gold label pad, and then ...

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Abstract

The invention discloses a colloidal gold method detection test strip and a reagent kit for IgM and IgG antibodies of mycoplasma pneumonia (MP) and a preparation method of the reagent kit. The test strip determines the IgM antibody and the IgG antibody by using a principle of an immunocapture method; the specific IgM and IgG antibodies of the MP can be detected jointly by one operation; the operation process is simplified; and the reagent kit is simple, convenient, rapid and accurate in detection, is particularly applicable to primary screening and epidemiological survey, and has an auxiliary diagnosis effect on early and interim mycoplasma pneumoniae infection.

Description

technical field [0001] The invention belongs to the field of rapid detection of colloidal gold immunochromatography, and relates to a test strip, kit and preparation technology for detecting specific IgM and IgG antibodies by immunoassay; A test strip, a test kit and a preparation method thereof for combined detection of mycoplasma pneumoniae IgM and IgG antibodies by chromogenic immunochromatographic reaction. Background technique [0002] Immunoglobulin M (Immunoglobulin M, IgM) and Immunoglobulin G (Immunoglobulin G, IgG) are the two most important antibodies in humans or animals. After an antigen (bacteria or virus, etc.) enters the body, the first immunoglobulin that appears is IgM antibody, followed by IgG antibody. Although IgA antibody may also appear in some cases, because its clinical significance is basically the same as that of IgG, and the IgA content is very low, it has little practical significance in clinical testing, so it is generally not distinguished in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/544
Inventor 不公告发明人
Owner JIANGSU KEYGEN BIOTECH CORP LTD
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