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 detection system for tick-borne encephalitis virus infection, and can solve problems such as the instability of Renilla luciferase

Active Publication Date: 2017-06-06
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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  • System for detecting tick-borne encephalitis virus infection
  • System for detecting tick-borne encephalitis virus infection
  • 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 to detect TBEV infection serum

[0103] This example 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 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 position of SEQ ID No.1 is the sequence of tick-borne encephalitis virus EIII protein, the 1st-199th position of SEQ ID No.1 is the sequence of Nanoluc, and the 200th-207th position of SEQ ID No.1 is The sequence of the linker peptide.

[0104] 1. Preparation of Nanoluc-EIII

[0105] 1.1 Construction of recombinant vector

[0106] Replace the sequence between the NheI and XhoI recognition sites of the pcDNA3.1(+) vector with the encoding gene of the fusion protein of the tick-borne encephalitis virus EIII...

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

Claims

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Application Information

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Patent Type & Authority Applications(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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