Toxoplasma gondii IgM antibody and total immunoglobulin combined detection reagent strip and its preparation method

A combined detection and Toxoplasma gondii technology, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of high false positives, low value, poor sensitivity of IgM antibody, etc., and achieve simple and fast detection, wide application and high sensitivity Effect

Inactive Publication Date: 2012-01-11
ZHONGSHAN HOSPITAL XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The diagnosis method of toxoplasmosis involves isolating toxoplasma directly from organs, blood, cerebrospinal fluid and other tissues for etiological diagnosis, which takes days or even weeks, is time-consuming and laborious, and has little value in practical applications
Routine clinical detection mainly uses various serological methods to detect its specific IgG and IgM ([3] Zhou Bin, Zhang Jue, Wang Ke, et al. The establishment and implementation of double-labeled time-resolved fluorescence immunoassay for Toxoplasma gondii IgG and IgM antibodies Its preliminary clinical application [J]. Chinese Journal of Laboratory Medicine, 2010, 33 (010): 957-959.), serological methods can diagnose congenital, acute and chronic toxoplasmosis, but because most immunocompromised people or The anti-toxoplasma IgG titer of animals does not rise or does not appear high IgM titer, so serological methods cannot be used effectively to diagnose toxoplasmosis in immunocompromised humans or animals
In addition, the serum antibody titer will gradually decrease after the antigen disappears for a long time, so it cannot be used to evaluate the therapeutic effect
At present, there are high false positives in the detection of IgG antibodies, and the general sensitivity of IgM antibody detection is poor ([4] Han Jingyun, Liu Qian, Guo Jian. Technical progress in laboratory diagnosis of Toxoplasma gondii infection [J]. Laboratory Medicine, 2009, 24(5): 393-395.)

Method used

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  • Toxoplasma gondii IgM antibody and total immunoglobulin combined detection reagent strip and its preparation method
  • Toxoplasma gondii IgM antibody and total immunoglobulin combined detection reagent strip and its preparation method
  • Toxoplasma gondii IgM antibody and total immunoglobulin combined detection reagent strip and its preparation method

Examples

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preparation example Construction

[0042] The preparation method of the combined detection reagent strip of the toxoplasma IgM antibody and the total antibody comprises the following steps:

[0043] 1) Preparation of Toxoplasma gondii recombinant antigens SAG1 (P30), SAG2 (P22), ROP2 and GRA7

[0044] Using gene cloning technology, PCR amplifies DNA encoding Toxoplasma gondii antigen, inserts it into Escherichia coli for expression, and obtains Toxoplasma gondii recombinant antigens SAG1 (P30), SAG2 (P22), ROP2 and GRA7.

[0045] 2) Spotting on nitrocellulose membrane

[0046] Coat anti-human IgM specific fragment μ chain monoclonal antibody on the detection line of Toxoplasma gondii IgM antibody, coat anti-human Ig monoclonal antibody at the total antibody detection line place, dry, the concentration of described anti-human Ig monoclonal antibody The concentration of anti-human IgM specific fragment μ chain monoclonal antibody can be 1-4 mg / mL, and the IgG antibody of goat anti-toxoplasma antigen SAG1 (P30), ...

Embodiment 1

[0071] The first reagent strip A is coated with anti-human IgM specific fragment μ chain monoclonal antibody on the nitrocellulose membrane (NC membrane) IgM detection line, and the second reagent strip B is coated with anti-human Ig monoclonal antibody on the total antibody detection line Antibodies, coated with goat anti-toxoplasma antigens SAG1 (P30), SAG2 (P22), ROP2 and GRA7 antibodies at the control line C, dried at room temperature, sealed and stored at room temperature for future use. Among them, the concentration of anti-human IgM specific fragment μ chain monoclonal antibody and anti-human Ig monoclonal antibody is 1 mg / mL, and goat anti-toxoplasma antigen (SAG1 (P30), SAG2 (P22), ROP2 and GRA7) IgG antibody is obtained by Goat anti-toxoplasma antigen (SAG1(P30), SAG2(P22), ROP2 and GRA7) IgG antibody consists of anti-SAG1(P30)-IgG antibody, anti-SAG2(P22)-IgG antibody, anti-ROP2-IgG antibody and anti-GRA7- IgG antibodies were mixed at a volume ratio of 1:1:1:1, and ...

Embodiment 2

[0075] Similar to Example 1, the difference is that the gold colloid pad and the total antibody detection line of Toxoplasma gondii are only composed of SAG1(P30), SAG2(P22), ROP2 and GRA7, and do not contain SAG1(P30), SAG2(P22), ROP2 and GRA7 . Result judgment is identical with embodiment 1.

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Abstract

A Toxoplasma gondii IgM antibody and total immunoglobulin combined detection reagent strip and its preparation method relate to a Toxoplasma gondii IgM antibody and total immunoglobulin combined detection reagent. A preparation method provided by the invention comprises the following steps of: arranging two reagent strips which are both equipped with a vector board, a sampling pad, a colloidal gold pad, a nitric acid fiber film, a contrast line and an absorption pad; arranging a Toxoplasma gondii IgM antibody detection line and a total immunoglobulin detection line on the two reagent strips; preparing Toxoplasma gondii recombinant antigen SAG1(P30), SAG2(P22), ROP2 and GRA7; carrying out spotting of the cellulose nitrate membrane; preparing colloidal gold; carrying out marking of colloidal gold with SAG1(P30), SAG2(P22), ROP2 and GRA7; and preparing the immunity chromatography detector strip. The detection reagent strip provided by the invention is used for the detection of Toxoplasma gondii IgM antibody and total immunoglobulin in whole blood, serum, blood plasma and cerebrospinal fluid specimen. The detection provided by the invention requires little amount of specimen and no special apparatus; the detection result is directly read by naked eyes; in addition, the detection is simple, rapid, accurate and reliable with strong specificity and high sensitivity, requires low cost, and is widely applied.

Description

technical field [0001] The invention relates to a combined detection reagent for toxoplasma IgM antibody and total antibody, in particular to a combined detection reagent strip for toxoplasma gondii IgM antibody and total antibody by colloidal gold immunochromatography (immunochromatography) and a preparation method thereof. Background technique [0002] Toxoplasmosis is a zoonotic parasitic disease caused by Toxoplasmagondii that seriously endangers human health. The pathogen Toxoplasma can parasitize in the nucleated cells of humans and various animals. and animals are generally susceptible. The disease is widely distributed all over the world. According to statistics, about 1 billion people in the world are infected by Toxoplasma gondii ([1] Xi Linlin, Li Wei. Research progress on pathogenic mechanism, virulence and genotype of Toxoplasma gondii[J]. Chinese Journal of Pathogenic Biology , 2009, 4(11): 859-861.). Toxoplasmosis is widely distributed in my country, and the...

Claims

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Application Information

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Patent Type & AuthorityApplications(China)
IPC IPC(8): G01N33/577G01N33/544G01N33/531
Inventor刘莉莉林丽蓉杨天赐张忠英张长弓
OwnerZHONGSHAN HOSPITAL XIAMEN UNIV