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Ekpoma virus detection kit and detection method thereof

A detection kit, Coupon heat technology, applied in the field of medical detection, can solve the problems of complex preparation methods, long time-consuming, expensive, etc., and achieve the effects of convenient and fast production, easy operation, high stability and specificity

Inactive Publication Date: 2018-07-24
李梅秀
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing detection kits for Kupo fever virus have problems such as complex preparation methods, long time-consuming, and high prices

Method used

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  • Ekpoma virus detection kit and detection method thereof
  • Ekpoma virus detection kit and detection method thereof
  • Ekpoma virus detection kit and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Human peripheral blood (PBMC) samples were derived from Kupo fever virus-infected patients and non-Kupuo virus-infected populations, and the number was 127 samples to be tested.

[0042] 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.

[0043] The detection steps of each sample to be tested are as follows:

[0044] (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;

[0045] (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 anti-hum...

Embodiment 2

[0054] Human peripheral blood (PBMC) samples were derived from Kupo fever virus-infected patients and non-Kupuo virus-infected populations, and the number was 127 samples to be tested.

[0055] 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.

[0056] The detection steps of each sample to be tested are as follows:

[0057] (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;

[0058] (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 anti-h...

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Abstract

The invention provides an Ekpoma virus detection kit and a detection method thereof. The kit comprises six polypeptides with amino acid sequences shown as SEQ ID NO.1 to SEQ ID NO.6. The detection method comprises the following steps: (1) separating a sample to be detected by separation liquid, and resuspending separated cells in a serum-free culture medium; (2) adding the six polypeptides with the amino acid sequences shown as SEQ ID NO.1 to SEQ ID NO.6 into the resuspended cells for co-incubation, then adding a high-specificity mouse anti-human IgG antibody into a co-incubation system for antigen and antibody reaction; (3) washing a system obtained after the antigen and antibody reaction, adding a developing substrate for developing reaction, and counting spots generated in the reaction.The kit provided by the invention can be artificially synthesized by full sequences and is convenient and quick for production, high in stability and specificity and easy and convenient to operate, so that 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 Kupo fever virus and a detection method thereof. Background technique [0002] Ekpoma virus (Ekpoma virus) is a virus newly discovered in Nigeria, West Africa in 2015. It is divided into two subtypes, type 1 (EKV1) and type 2 (EKV2). reports. At the same time, the virus is highly homologous to another Bas-congo virus that causes hemorrhagic fever and has a high fatality rate. Due to the small number of cases, the shape of the pathogenic organism is unknown. Fever manifests as normal body temperature in the morning, low fever in the afternoon, high fever to 40°C in the morning, and fluctuations in body temperature in the morning and evening. Culicoides are small blood-sucking insects that live in shrubs and are easy to spread zoonotic diseases. Because Culicoides spread and body temperature fluctuates, we named it Kupo fever in Chinese....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983G01N2333/005
Inventor 李梅秀
Owner 李梅秀
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