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A system for detecting tick-borne encephalitis virus infection

A technology for tick-borne encephalitis virus and encephalitis virus, which is applied in the field of systems for detecting tick-borne encephalitis virus infection, and can solve problems such as instability of renilla luciferase

Active Publication Date: 2020-06-05
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Renilla luciferase (Renilla luciferase, RLuc) reporter gene, with a full length of 936bp, is expressed in a eukaryotic expression system to produce Renilla luciferase, which can be detected by interacting with the Renilla luciferase substrate to emit bioluminescence Renilla luciferase activity, its detection sensitivity is extremely high, but the currently used Renilla luciferase is unstable

Method used

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  • A system for detecting tick-borne encephalitis virus infection
  • A system for detecting tick-borne encephalitis virus infection
  • A system for detecting tick-borne encephalitis virus infection

Examples

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Embodiment 1

[0102] Embodiment 1, fusion protein Nanoluc-EIII can be used for detecting TBEV infection serum

[0103] This embodiment provides the application of the fusion protein of tick-borne encephalitis virus EIII protein and Nanoluc in detecting TBEV infected serum, the fusion protein of tick-borne encephalitis virus EIII protein and Nanoluc is named as Nanoluc-EIII, and Nanoluc-EIII is SEQ The coding sequence of the protein shown in ID No.1 is shown in SEQ ID No.2. Wherein the 208-300th of SEQ ID No.1 is the sequence of tick-borne encephalitis virus EIII protein, the 1-199th of SEQ ID No.1 is the sequence of Nanoluc, and the 200-207th of SEQ ID No.1 is Sequence of linker peptide.

[0104] 1. Preparation of Nanoluc-EIII

[0105] 1.1 Construction of recombinant vector

[0106] The sequence between the NheI of pcDNA3.1 (+) vector and the XhoI recognition site is replaced with the coding gene of the fusion protein of tick-borne encephalitis virus EIII protein and Nanoluc (the nucleos...

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Abstract

The invention discloses a system for detecting tick-borne encephalitis virus infection. The system for detecting tick-borne encephalitis virus infection consists of a fusion protein composed of a tick-borne encephalitis virus antigen and luciferase NanoLuc and other reagents and / or instruments utilizing luciferase immune co-precipitation technology to detect whether infection of the tick-borne encephalitis virus occurs. The fusion protein is the protein of following I1) or I2): I1) a protein shown as SEQ ID No.1; and I2) a I1) derived protein that is obtained by substitution and / or deletion and / or adding of one or more amino acid residues in the amino acid sequence of SEQ ID No.1 and has the same functions. Experiments prove that the Nanoluc-EIII and the system for detecting tick-borne encephalitis virus infection can detect whether human serum is infected with tick-borne encephalitis virus at 37DEG C and room temperature.

Description

technical field [0001] The invention relates to a system for detecting tick-borne encephalitis virus infection in the field of biotechnology. Background technique [0002] TBEV is a virus transmitted by tick bites, which can cause encephalitis and meningitis, and can cause severe neurological sequelae. The diagnosis of TBEV infection mainly relies on serology and pathogenic nucleic acid detection. The encoded proteins of TBEV include: C, PrM(M), E, ​​NS1, NS2a, NS2b, NS3, NS4a, NS4b, NS. [0003] The luciferase-based immunocapture system (Luciferase immunocapture, Luc-IC) recombinedly expresses the detection antigen and luciferase, and after incubating with the sample to be tested, determines the antibody content in the sample to be tested by detecting the intensity of luciferase , has the characteristics and advantages of being fast and sensitive. The advantage of the Luc-IC detection system is that it chooses to express antigens in mammalian cells. The protein expressed...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00G01N33/68G01N33/577G01N33/569C12N15/62
Inventor 杨银辉康晓平霍耐凡李裕昌吴晓燕张雨李靖户义
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
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