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A kit for detecting human fever with thrombocytopenia syndrome virus antibody igg and its application

A platelet and syndrome technology, which is used in the field of kits for detecting human fever with thrombocytopenia syndrome virus antibody IgG, can solve the problem of inability to evaluate the protective effect of the vaccine on the level of previous infection in the population

Active Publication Date: 2015-11-18
ZHENGZHOU IMMUNO BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the virus sequence has been elucidated, and the detection standard based on RT-PCR has been established, but the immunological detection reagents for SFTS have not yet been perfected, so that the evaluation of the virus infection lacks a confirmation method for PCR results, and it is impossible to evaluate the past infection of the population level and vaccine protection

Method used

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Comparison scheme
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Embodiment Construction

[0015] The concrete preparation method of kit of the present invention is as follows:

[0016] 1. Expression and purification of rSFTSN protein

[0017] Referring to the sequence in NCBI (4J4R_A), the reading frame of its N protein was synthesized and the restriction site NdeI was added at the 5', and the restriction site XhoI was added at the 3'. Using conventional molecular biology methods, the reading frame was inserted between the bacterial expression plasmid PET30A (novagen) and the bacterial expression plasmid PET30A-rSFTSN. The plasmid was routinely transformed into E.ColiBL21DE3 (novagen), and rSFTSN recombinant protein was induced to express according to the method recommended by the manufacturer, and the rSFTSN recombinant protein was routinely purified with Hitrap (novagen). The molecular weight and purity of the protein were detected by SDS-PAGE, and routine protein quantification was used for future use.

[0018] 2. Expression and purification of rRVFN protein...

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Abstract

The invention discloses a kit for detecting human severe fever with thrombocytopenia syndrome (SFTS) virus antibody IgG. The kit comprises a reaction titration microplate and a detection reagent containing a negative control, a positive control, a sample diluent, HRP mouse anti-human IgG, a substrate solution, a color-developing reagent and a stop solution, wherein the reaction titration microplate is coated with rSFTSN proteins with the concentration 0.2-0.4 microgram / ml; the sample diluent contains rRVFN recombinant proteins with the concentration of 5-10 microgram / ml; an HRP mouse anti-human IgG enzyme contains rRVFN recombinant proteins with the final concentration of 5-10 microgram / ml. The invention further discloses application of the kit to preparation of a diagnostic reagent for observing epidemic diseases infecting human beings and evaluating antibody levels after vaccine immunity. When the kit is used for detecting the human SFTS virus antibody IgG, the sensitivity can be up to 99.8 percent, the in-analysis accuracy is 2.5 percent (n=10), the inter-analysis accuracy is 4.5 percent (n=10), and the specificity is 99.5 percent.

Description

Technical field [0001] The present invention involves the field of biotechnology, especially a kit that involves a test of a detection person's heating with platelets to reduce the syndrome of the syndrome.Application in the diagnostic reagent of the evaluation. Background technique [0002] Bunyaviride is a type of negative chain RNA virus with envelope. At present, there are at least 350 viruses. They are the most virus in the insecticadia virus, including cloth that can infect people and animals.Orthobunyavirus, PHLEBOVIRUS, Nairovirus, Hantavirus, and TOSPOVIRUS (Hantavirus).Among them, the White Burdock virus is currently recognized as two major groups internationally internationally, one group is the pathogenesis of the pathogenesis (SandflyfeverGroup), such as RiftVallyfevervirus, and the other group.The UUKU-NIEMIGROUP generally does not cause disease to humans, such as UUKUNIE-MIVIRUS.In recent years, Henan, Jiangsu, Hubei, Anhui, Shandong, Liaoning, Zhejiang and other p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/535
CPCG01N33/535G01N33/56983G01N33/6893G01N2333/175
Inventor 赵巧辉黄学勇许汴利杜燕华李桂林付光宇吴学炜
Owner ZHENGZHOU IMMUNO BIOTECH