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Kit for detecting trypanosoma cruzi antibody as well as preparation and application thereof

A technology of Trypanosoma cruzi and a kit, which is applied in biological testing, material inspection products, and analysis through chemical reactions of materials, and can solve problems such as limited detection sensitivity, non-specific binding, and poor antigen specificity , to achieve the effects of shortening the test time, improving binding specificity, specificity and sensitivity

Active Publication Date: 2016-05-04
SHENZHEN NEW INDS BIOMEDICAL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the chemiluminescence method using the double-antigen sandwich method to detect chagas, most of the antigens that can be paired with each other have poor specificity and are prone to non-specific binding, resulting in limited detection sensitivity.

Method used

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  • Kit for detecting trypanosoma cruzi antibody as well as preparation and application thereof
  • Kit for detecting trypanosoma cruzi antibody as well as preparation and application thereof
  • Kit for detecting trypanosoma cruzi antibody as well as preparation and application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0048] The following is the preparation method of each component of the kit:

[0049] Preparation 1: Preparation of magnetic sphere suspension coated with chagas antigen

[0050] The immunomagnetic beads used in this preparation step are selected from the nano-magnetic microsphere suspension produced by Merck Company with a concentration of 100 mg / mL and a hydroxyl group of 95 mgKOH / g.

[0051] (1) Preparation of buffer:

[0052] Weigh 2.55g of sodium acetate trihydrate, dissolve it in 4500mL of purified water, add 14mL of acetic acid, and mix well to obtain an acetate buffer solution with a pH of 3.6.

[0053] (2) Reaction of magnetic balls coated with chagas antigen:

[0054] Suspend the magnetic microspheres in the above-mentioned pH3.6 acetate buffer solution 5 times the volume of the coating, so that the concentration of the magnetic beads is 20 mg / mL; then add 1-cyclohexyl-2-morpholinoethylcarbodiimide to Add tosylate (CMC) to a concentration of 10 mg / mL; add the chag...

Embodiment 1

[0090] In order to achieve the above object, the present invention provides embodiment 1, its kit components and preparation are as follows:

[0091] According to the above-mentioned preparation 1, a suspension of magnetic spheres coated with chagas antigen was prepared;

[0092] The chagas secondary antibody labeled with ABEI was prepared according to the above preparation 4;

[0093] Prepare the low-point calibrator and high-point calibrator for the chagas antibody as described in Preparation 5 above.

[0094] Embodiment 1 adopts the method of direct connection coating or labeling, which is simple to operate and easy to prepare.

Embodiment 2

[0095] In order to further improve the detection effect of the kit, the present invention provides preferred embodiment 2, its components and preparation are as follows:

[0096] According to the above-mentioned preparation 2, a suspension of magnetic spheres coated with SA was prepared;

[0097] Prepare biotinylated chagas antigen according to Preparation 3 above;

[0098] Mix the product obtained in the above two steps according to the weight ratio of the magnetic ball suspension and the chagas antigen at 200:3, put it in a constant temperature shaking water bath at 37°C for 24 hours, wash it several times and resuspend it to obtain an indirect coating Magnetic sphere suspension with chagas antigen.

[0099] The chagas secondary antibody labeled with ABEI was prepared according to the above preparation 4;

[0100] Prepare the low-point calibrator and high-point calibrator for the chagas antibody as described in Preparation 5 above.

[0101] SA and biotin in this example c...

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Abstract

The invention provides a kit for detecting a trypanosoma cruzi antibody and a preparation method of the kit. The kit comprises a component A and a component B, wherein the component A is a chagas antigen which is marked by a tracking marker or coated with a magnetic ball; the component B is a chagas second antibody which is coated with the magnetic ball or marked by the tracking marker; and any one of the component A and the component B is marked by the tracking marker and the other one is coated with the magnetic ball. The invention further provides a method for detecting the chagas antibody. By utilizing the kit provided by the invention, the concentration of chagas in a sample can be accurately and flexibly determined. On the basis, a pre-treatment solution is used for carrying out pre-treatment optimization so that the background interference can be reduced and the detection flexibility is improved. Meanwhile, detection of the full-automatic chemiluminiscence method can be realized with the help of a chemiluminiscence immunoassay analyzer, so that the operation time is shortened and manual operation errors are reduced.

Description

technical field [0001] The invention relates to the technical field of biological substance detection, in particular to a kit for detecting trypanosoma cruzi antibodies and its preparation, and a method for detecting trypanosoma cruzi antibodies using the kit. Background technique [0002] Trypanosomiasis, also known as American trypanosomiasis / Chagas, is a zoonotic parasitic protozoan disease caused by Trypanosomacruzi. The pathogen is Trypanosoma cruzi, which is a single-celled protozoan that can grow in different forms in the blood, cells, and digestive tracts of mammals (including humans), and the epidemic area is mainly passed through "patient or storage host → trypanosoma". Stinkbug → human" route of transmission. Pathogens in non-endemic areas can be transmitted through blood products, organ transplants, vertical transmission or eating contaminated food. [0003] The clinical symptoms of the disease vary with different stages of infection. About 95% of patients in t...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N21/76
CPCG01N21/76G01N33/68
Inventor 饶微刘坤方中刚刘晓森李婷华
Owner SHENZHEN NEW INDS BIOMEDICAL ENG
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