Toxoplasma gondii detection kit

A technology for detecting kits and Toxoplasma gondii, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of low specificity, differences, false negatives, etc., and achieve the effect of overcoming the poor detection rate

Inactive Publication Date: 2015-09-09
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The appearance and disappearance of IgM are faster than IgG, and due to the interference of disease factors such as rheumatism, the current commercial IgM detection kits have the disadvantage of low specificity
After being infected with Toxoplasma gondii, the production of antibodies varies significantly due to individual differences, and even a large proportion of false negatives occurs, so the detection efficiency of antibodies is not high, so it is urgent to develop a method for detecting Toxoplasma gondii with high sensitivity and specificity. new method

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Toxoplasmosis Detection Kit Based on IFN-γ Release

[0028] The kit includes the following components:

[0029]

[0030]

[0031] In addition, the routine reagents and consumables that need to be prepared include PBS, distilled water, double distilled water, six-well cell culture plates, and tips of various specifications.

Embodiment 2

[0032] Example 2 The use procedure of the toxoplasmosis detection kit released based on IFN-γ

[0033] 1. Sample pretreatment

[0034] (1) Peripheral blood collection: Aseptically collect human or animal anticoagulant blood with vacuum blood collection tubes, 3ml for each subject.

[0035] (2) Peripheral blood dilution: within 12 hours after collection, add an equal volume of cell culture medium (containing 90% RPIM1640 medium, 10% fetal bovine serum, 100 units / ml of penicillin and streptomycin, 0.0002% β-mercaptoethanol) to dilute and mix.

[0036] (3) Cell plating: Add the diluted blood to a six-well cell culture plate, 2ml per well, and plate 3 wells of blood cells from each subject.

[0037] (4) Stimulation with Toxoplasma gondii antigen: Add cell culture medium to the first well as a negative control, add Toxoplasma antigen at a final concentration of 12 μg / ml to the second well, and add concanavalin A at a final concentration of 20 μg / ml to the third well as a positiv...

Embodiment 3

[0055] Example 3 Detection of IFN-γ secreted by specific T cells in human whole blood by ELISA to detect whether people are infected with Toxoplasma gondii

[0056] Firstly, human toxoplasmosis IgG detection kit was used to detect serum IgG of people under different infection states; secondly, human anticoagulant blood was collected, stimulated with toxoplasma antigen, and the supernatant was taken for quantitative detection of IFN-γ after 48 hours.

[0057] It was found that using ELISA to detect IFN-γ secreted from human whole blood is a new method to detect whether people are infected with Toxoplasma gondii, suggesting that this method can be used to detect human toxoplasmosis, and this method can detect human toxoplasmosis The state of cellular immune response in the human body after Toxoplasma gondii infection.

[0058] (1) Detection of IgG levels in human serum:

[0059] Human serum was collected, and IgG levels in the serum of 12 subjects were detected with a human tox...

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Abstract

The invention provides a toxoplasma gondii detection kit, and is characterized in that the kit includes a toxoplasma gondii antigen, an IFN-gamma capture antibody, an IFN-gamma standard substance, a biotin-labeled IFN-gamma detection antibody, a cell culture medium, a positive control, a negative control, a coating liquid, a confining liquid, an HRP-labeled streptavidin, a developing solution and a stop solution. The kit has high sensitivity and specificity, and overcomes common shortcomings of poor detection rate and high misdiagnosis rate of a conventional toxoplasma gondii antibody detection kit.

Description

technical field [0001] The invention relates to the technical field of biological quarantine, in particular to a detection kit for toxoplasma gondii. Background technique [0002] Toxoplasma gondii is an obligate intracellular parasite that infects the nucleated cells of almost all warm-blooded animals. Causes widespread zoonotic parasitic diseases worldwide. It is estimated that approximately one-third of the world's population is infected with the parasite. Toxoplasma gondii is an opportunistic pathogenic parasite that mainly harms pregnant women and people with weakened immune systems. Infection by people with normal immune functions does not cause obvious clinical symptoms. [0003] Toxoplasma gondii infection can induce the body to produce a strong Th1-type cellular immune response, leading to specific CD4+T and CD8+T cells secreting a large amount of IFN-γ to resist Toxoplasma gondii infection. [0004] At present, enzyme-linked immunosorbent assay (ELISA) is the mo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/6866G01N33/56905
Inventor 索勋刘贤勇尹青
Owner CHINA AGRI UNIV
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