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Novel Bunia virus detection kit and detection method thereof

A bunya virus, detection kit technology, applied in the field of medical detection, can solve the problem of high mortality, achieve the effect of reducing cost, high stability and specificity, controllability and accuracy

Inactive Publication Date: 2018-07-27
李梅秀
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] People infected by the new bunyavirus often have acute onset and high mortality, but the current treatment for the new bunyavirus infection is mainly symptomatic and supportive treatment

Method used

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  • Novel Bunia virus detection kit and detection method thereof
  • Novel Bunia virus detection kit and detection method thereof
  • Novel Bunia virus detection kit and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040]Human peripheral blood (PBMC) samples came from patients infected with the new bunyavirus and people who were not infected with the new bunyavirus, and the number was 127 samples to be tested.

[0041] 10 mL of blood was collected from each individual with heparin sodium collection tubes, and transported to the laboratory within 6 hours at room temperature.

[0042] The detection steps of each sample to be tested are as follows:

[0043] (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;

[0044] (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 Specifi...

Embodiment 2

[0053] Human peripheral blood (PBMC) samples came from patients infected with the new bunyavirus and people who were not infected with the new bunyavirus, and the number was 127 samples to be tested.

[0054] 10 mL of blood was collected from each individual with heparin sodium collection tubes, and transported to the laboratory within 6 hours at room temperature.

[0055] The detection steps of each sample to be tested are as follows:

[0056] (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;

[0057] (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 Specif...

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Abstract

The invention provides a novel Bunia virus detection kit and a detection method thereof. The novel Bunia virus detection kit comprises six polypeptides with amino acid sequences shown in SEQ ID NO.1 to SEQ ID NO.6. The detection method comprises the following steps of (1) separating a to-be-detected sample by a separating liquid, and re-suspending the separated cells in a serum-free culture medium; (2) adding the six polypeptides with amino acid sequences shown in SEQ ID NO.1 to SEQ ID NO.6 into the re-suspended cells, hatching together, and adding a mouse anti-human IgG antibody with high specificity into the co-hatched system to perform antigen and antibody reaction; (3) washing the system after antigen and antibody reaction, adding a color developing primer to develop color, and counting the number of spots which are generated by reaction. The novel Bunia virus detection kit has the advantages that the artificial whole-sequence synthesizing effect is realized; the production is convenient and rapid; the stability and specificity are high; the operation is simple and convenient; the cost is greatly reduced.

Description

technical field [0001] The invention belongs to the technical field of medical detection, and in particular relates to a detection kit for a novel bunyavirus and a detection method thereof. Background technique [0002] The new bunyavirus is a virus isolated by the health department from tick poisoning patients. Fever with thrombocytopenia syndrome caused by a new type of bunyavirus infection in humans is a new infectious disease with fever accompanied by leukopenia, thrombocytopenia and multiple organ dysfunction as the main clinical manifestations. After infection with the new bunyavirus, it can rapidly cause damage to the liver and kidney function of the patient, causing the patient to die due to disseminated intravascular coagulation, shock, and respiratory failure, with a mortality rate as high as 30%. Epidemiological studies have shown that the distribution of new bunyaviruses is extremely wide: since the first successful isolation and identification of new bunyavirus...

Claims

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Application Information

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IPC IPC(8): G01N33/569
CPCG01N33/56983G01N2333/175G01N2469/10
Inventor 李梅秀
Owner 李梅秀
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