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A colloidal gold test strip for detecting toxoplasma gondii antibody

A technology of colloidal gold test paper and Toxoplasma gondii, which is applied in the field of serological detection, can solve problems such as death and serious disease, and achieve the effects of strong specificity and sensitivity, simple operation and less sample amount required.

Inactive Publication Date: 2018-01-02
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most hosts infected with Toxoplasma gondii usually show relatively mild symptoms, but if the body's immunity is weakened, it can lead to serious illness and even death

Method used

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  • A colloidal gold test strip for detecting toxoplasma gondii antibody
  • A colloidal gold test strip for detecting toxoplasma gondii antibody
  • A colloidal gold test strip for detecting toxoplasma gondii antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1, TgGRA1 protein preparation

[0042] 1. Cloning of TgGRA1 gene

[0043] Primers were designed to amplify the TgGRA1 gene, and the primers are shown in Table 1 below. The two ends of the primers contain enzyme cutting sites, which are indicated in lowercase. The 5' end of the primer has added protective bases, and the 3' end is the TgGRA1 gene sequence.

[0044] Table 1 is the primers used to amplify the TgGRA1 gene

[0045]

[0046] Using the cDNA of the Toxoplasma gondii RH strain as a template, PCR amplification was performed with the upstream and downstream primers of the TgGRA1 gene to obtain a 495bp PCR product, which was the TgGRA1 gene. After sequencing, the nucleotide sequence of the TgGRA1 gene was sequence 1 in the sequence table, The protein TgGRA1 encoded by the TgGRA1 gene, the amino acid sequence of the protein TgGRA1 is sequence 2 in the sequence listing;

[0047] Above-mentioned reaction system is as follows table 2:

[0048] Table 2 ...

Embodiment 2

[0074] Embodiment 2, the preparation of colloidal gold test paper

[0075] 1. Preparation of nitrocellulose membrane containing detection line T and quality control line C

[0076] 1), preparation of coated antigen

[0077] TgGRA1 protein solution is to dissolve the tagged TgGRA1 protein prepared in Example 1 above in PB (PB preparation: weigh 5.37 g of disodium hydrogen phosphate, 0.2 g of potassium chloride, and 0.2 g of potassium dihydrogen phosphate, add ultrapure water to 1000ml, mix well and set aside.), and the concentration of the tagged TgGRA1 protein is 2mg / ml, which is the TgGRA1 antigen.

[0078] Pig IgG protein solution is to dissolve the above-mentioned pig IgG protein (purchased from Zhongke Maichen (Beijing) Technology Co., Ltd., catalog number: SP7-10) in PB, and the concentration of pig IgG protein is 1mg / ml, which is the pig IgG protein solution. IgG antigen.

[0079] 2), coating

[0080] A detection line (T line) and a quality control line (C line) were...

Embodiment 3

[0107] Embodiment 3, the detection effect of test strip

[0108] 1. Specificity and sensitivity

[0109] The colloidal gold test strip (GICA) and indirect immunofluorescence test (IFAT) two methods prepared by above-mentioned embodiment 2 detect identical rabbit, dog, cat serum, compare the coincidence rate of these two kinds of method results, calculate test strip Based on the specificity and sensitivity of IFAT.

[0110] 1) Colloidal gold test strip detection

[0111] Add 100 μl of 20-fold diluted rabbit, dog, and cat serum samples to the sample pad of the colloidal gold test strip prepared in 3 above, and judge the result within 15 minutes. according to image 3 Judgment method to determine whether toxoplasma infection.

[0112] 2) The operation steps of the IFAT method are as follows:

[0113] A. Collect the freshly released Toxoplasma gondii, suspend them with an appropriate amount of PBS, and drop them on the glass slides with adsorption holes. After being adsorbed...

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Abstract

The invention discloses a colloidal gold test strip for testing toxoplasma gondii antibodies.The colloidal gold test strip comprises a bottom board, a sample pad, a gold-labeled protein-enveloped conjugate pad, a nitrocellulose membrane containing a detection line T and a quality control line C and an absorbent pad, wherein the sample pad, the conjugate pad, the nitrocellulose membrane and the absorbent pad are disposed on the bottom board.Experiments prove that the colloidal gold test strip is capable of detecting serum antibodies of toxoplasma gondii in animal bodies and achieving differentiating diagnosis of protozoan infection of animals conveniently, cost saving and simple and convenient to operate and is applicable to the animals such as pigs, dogs and cats.

Description

technical field [0001] The invention belongs to the field of serological detection and relates to a colloidal gold test strip for detecting toxoplasma gondii antibody. Background technique [0002] Toxoplasmosis is caused by the obligate intracellular parasitic protozoan Toxoplasmagondii (T. Toxoplasma gondii can infect a variety of animals and humans, and the occurrence, symptoms, and pathogenicity of the infection are closely related to the genotype of Toxoplasma gondii, the amount of infection, the genetic background and immune status of the host. Most hosts infected with Toxoplasma gondii usually show relatively mild symptoms, but if the body's immunity is weakened, it can lead to serious illness and even death. Another hazard of toxoplasmosis is that it causes reproductive disorders such as abortion and stillbirth in pregnant animals. [0003] The diagnostic methods for Toxoplasma gondii infection and pathogenesis include etiology, serology, molecular biology and immu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/558G01N33/52
CPCG01N33/52G01N33/558G01N33/56905G01N2333/45
Inventor 刘群南汇珠刘晶
Owner CHINA AGRI UNIV
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