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Artificial antigen and kit for combined detection of eb virus rta protein antibody and eb virus early antigen ea antibody

A technology of Epstein-Barr virus and early antigen, applied in the field of medical immunology, in vitro serological diagnosis, and biotechnology, can solve the problems of poor sensitivity, poor specificity, and high missed detection rate of nasopharyngeal carcinoma, and achieve low cost, easy operation, and shortened time. The effect of detection time

Active Publication Date: 2017-03-08
同昕生物技术(北京)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A large number of clinical experiments have confirmed that the EB virus VCA-IgA antibody for the diagnosis of nasopharyngeal carcinoma has high sensitivity, but the specificity is poor, and it has a high detection rate in healthy people; the EB virus EA-IgA antibody is specific for the diagnosis of nasopharyngeal carcinoma Good sensitivity, but poor sensitivity, high missed detection rate of nasopharyngeal carcinoma

Method used

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  • Artificial antigen and kit for combined detection of eb virus rta protein antibody and eb virus early antigen ea antibody
  • Artificial antigen and kit for combined detection of eb virus rta protein antibody and eb virus early antigen ea antibody
  • Artificial antigen and kit for combined detection of eb virus rta protein antibody and eb virus early antigen ea antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Tandem expression of Epstein-Barr virus Rta protein and EA antigen in Escherichia coli

[0047] Look up the amino acid sequence of Epstein-Barr virus Rta protein, EA antigen (see sequence table SEQ ID NO:7 and SEQ ID NO:8) on NCBI website, adopt http: / / www.imtech.res.in / raghava / bcepred / bcepred_submission. html analyzes its epitope, selects its epitope, obtains the optimized Epstein-Barr virus Rta protein fragment (Seq ID No: 1) and the optimized Epstein-Barr virus EA antigen (Seq ID No: 2), and codes sub for optimization. The optimized Rta and EA antigen gene sequences were combined and synthesized, and primers were designed with Primer5.0 primer design software.

[0048] Upstream primer P1: 5'-CCTCGAGATGGCTTCTTCTAACCGT-3',

[0049] Downstream primer P2: 5'-CTGGATCCTTTGGTTTTGAAACGCAG-3'.

[0050] P1 and P2 primers respectively contain Xho I and BarmH I restriction sites.

[0051] The above synthetic gene was amplified by PCR technique.

[0052] PCR reacti...

Embodiment 2

[0057] Example 2 Preparation of a kit for joint detection of Epstein-Barr virus Rta protein antibody and early antigen EA antibody

[0058] (1) Composition of the kit

[0059] 1. ELISA plate coated with Rta-EA antigen: the coating protein is high-purity recombinant Rta-EA protein, and the coating concentration can be 0.05-0.25 μg / ml;

[0060] 2. The working dilution of HRP-labeled goat anti-human IgA can be 1:5000—1:80000;

[0061] 3. Calibrator solution, positive control solution, negative control solution, 1 bottle each, 1ml / bottle;

[0062] 4. Sample diluent: 50mM phosphate buffer containing 0.3M NaCl, 25ml / bottle, 2 bottles;

[0063] 5. Substrate chromogenic solution: composed of A liquid and B liquid, the substrate chromogenic liquid A is carbamide peroxide, 7ml / bottle, 1 bottle; the substrate chromogenic liquid B is tetramethylbenzidine, 7ml / bottle, 1 bottle;

[0064] 6. Termination solution: 2mol / l sulfuric acid;

[0065] 7. Concentrated washing solution: pH valu...

Embodiment 3

[0081] Embodiment 3 Clinical trial evaluation of kit of the present invention

[0082] 55 normal human samples and 58 patient samples diagnosed with nasopharyngeal carcinoma were detected with the kit prepared by the present invention. The detection procedure and result judgment were carried out in strict accordance with the usage method of the kit provided in Example 2, and VCA antibody was selected for detection at the same time. The kit and the EA antibody detection kit (both commercially purchased from the Chinese Institute of Preventive Medicine and Virology) were tested in parallel, and the specific operations were performed according to the instructions of the kits. The test results are shown in Table 1.

[0083] Table 1 Comparison of test results

[0084]

[0085] It can be seen from the above results that the kit provided by the present invention has excellent performance in the diagnosis and screening of nasopharyngeal carcinoma, with a sensitivity of 91.4% and a...

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Abstract

The invention relates to the technical fields of biotechnology, medical immunology and in vitro serological diagnosis, and particularly relates to an artificial antigen and a kit for joint detection of an Rta protein antibody of an epstein-barr (EB) virus and an early antigen ethyl acrylate (EA) antibody of the EB virus. The Rta protein antibody of the EB virus and the early antigen EA antibody existing in a to-be-detected sample can be accurately, efficiently, sensitively and specifically detected by the artificial antigen provided by the invention. By adopting the kit prepared from the artificial antigen, compared with the traditional detection method, the sensitivity and the specificity of nasopharynx cancer diagnosis are improved, the detection time is shortened, a patient can be timely treated, and the pain is relieved.

Description

technical field [0001] The invention relates to the technical fields of biotechnology, medical immunology and in vitro serological diagnosis, in particular to an artificial antigen and a kit for joint detection of Epstein-Barr virus Rta protein antibody and EB virus early antigen EA antibody. Background technique [0002] Epstein-Barr virus is the first time that Epstein and Barr successfully established Burkitt African children's lymphoma cells through in vitro suspension culture in 1964, and herpes virus particles were observed by electron microscope in the cell smears of the established strain. The virus is one of the causes of various malignant tumors (such as nasopharyngeal carcinoma), and it mainly infects epithelial cells and B lymphocytes in the human oropharynx. Most of the patients with nasopharyngeal carcinoma in southern China have detected the presence of Epstein-Barr virus genome. Nasopharyngeal carcinoma (nasopharygeal carcinoma, NPC) refers to malignant tumo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/70G01N33/68C12R1/93
Inventor 焦守恕李全贾凤芹
Owner 同昕生物技术(北京)有限公司
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