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Detection kit for American trypanosomiasis and detection method thereof

A detection kit, the technology of American trypanosoma, applied in the field of medical detection, to achieve the effect of simplified process, convenient and fast detection, and cost reduction

Inactive Publication Date: 2018-11-23
李梅秀
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no effective and accurate detection method for American trypanosomiasis

Method used

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  • Detection kit for American trypanosomiasis and detection method thereof
  • Detection kit for American trypanosomiasis and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Horse peripheral blood samples came from domestic horses infected with American trypanosomiasis and non-infected domestic horses, and the number was 127 samples to be tested.

[0044] 10 mL of blood was collected from each sampled individual with a sodium heparin collection tube, and transported to the laboratory within 6 hours at room temperature.

[0045] The detection steps of each sample to be tested are as follows:

[0046] (1) Separate the PBMC with the separation medium, resuspend the separated cells in serum-free medium, and the cell concentration is 1.5×10 5 Cells / mL; Add the resuspended cells into a PVDF96-well plate, add 100 μL to each reaction well, add each sample to 10 wells, and make 10 repetitions;

[0047] (2) Add 50 μL of a mixture consisting of serum-free medium and 6 polypeptides with a final concentration of 5 μM to each reaction well, mix well and carry out co-incubation; set a positive control and a negative control, and add high Specific mouse an...

Embodiment 2

[0056] Horse peripheral blood samples came from domestic horses infected with American trypanosomiasis and non-infected domestic horses, and the number was 127 samples to be tested.

[0057] 10 mL of blood was collected from each sampled individual with a sodium heparin collection tube, and transported to the laboratory within 6 hours at room temperature.

[0058] The detection steps of each sample to be tested are as follows:

[0059] (1) Separate the PBMC with the separation medium, resuspend the separated cells in serum-free medium, and the cell concentration is 3.5×10 5 Cells / mL; Add the resuspended cells into a PVDF96-well plate, add 100 μL to each reaction well, add each sample to 10 wells, and make 10 repetitions;

[0060] (2) Add 50 μL of a mixture consisting of serum-free medium and 6 polypeptides with a final concentration of 5 μM to each reaction well, mix well and carry out co-incubation; set a positive control and a negative control, and add high Specific mouse ...

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Abstract

The invention provides a detection kit for American trypanosomiasis and a detection method thereof. The kit comprises 6 polypeptides with amino acid sequences of SEQ ID NO. 1 to SEQ ID NO. 6. The detection method comprises the following steps: (1) separating a sample to be detected by utilizing a separation solution and re-suspending separated cells in a serum free medium; (2) adding 6 polypeptides with the amino acid sequences of SEQ ID NO. 1 to SEQ ID NO. 6 into the re-suspended cells and carrying out common incubation; then adding a high-specificity mouse anti-human IgG antibody into a common incubation system and carrying out antigen and antibody reaction; (3) washing a system subjected to the antigen and antibody reaction; then adding a chromogenic substrate and carrying out chromogenic reaction; counting the quantity of spots generated by the reaction. The kit provided by the invention can be obtained by artificial complete sequence synthesis, is convenient and rapid to produce and has high stability and specificity; the kit is simple and convenient to operate and the cost is extremely reduced.

Description

technical field [0001] The invention belongs to the technical field of medical detection, and in particular relates to a detection kit for American trypanosomiasis and a detection method thereof. Background technique [0002] American trypanosomiasis disease is caused by Trypanosoma cruzi, and its main transmission medium is the blood-sucking bug. It was discovered by Dr. Chagas in 1908, so it is also called Chagas disease. It is mainly prevalent in Central and South America, so it is called American trypanosomiasis. In the acute stage of the disease, there are fever, facial edema, lymphadenitis, and anemia. In the chronic phase, myocarditis, heart failure, giant esophagitis, megacolon, pulmonary and cerebral embolism, and sudden death often occur. [0003] Trypanosoma cruzi parasitizes in the blood and various tissue cells of humans and mammals. There are two forms of tryomastigotes and amastigotes in its life history. Tryomastigotes invade human blood through skin woun...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/569
CPCG01N33/6893G01N33/56905G01N2410/00
Inventor 李梅秀
Owner 李梅秀
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