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Chemiluminescence kit for detecting EB virus Rta/IgG antibody and use thereof

A chemiluminescence reagent, EB virus technology, applied in the fields of biotechnology, medical immunology and in vitro serological diagnosis, can solve the problems of enhanced background luminescence, limited application and development of sensitivity, elevated measurement background, etc., to simplify steps and shorten the Detection time, easy operation effect

Active Publication Date: 2015-11-11
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

Luminol, isoluminol and their derivatives are the earliest chemiluminescent substances used, but their application in chemiluminescence immunoassay requires the use of catalysts and enhancers, which will lead to enhanced background luminescence and increase the measurement background. High, which limits the sensitivity of this technique and its application and development

Method used

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  • Chemiluminescence kit for detecting EB virus Rta/IgG antibody and use thereof
  • Chemiluminescence kit for detecting EB virus Rta/IgG antibody and use thereof
  • Chemiluminescence kit for detecting EB virus Rta/IgG antibody and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1. Preparation of kit of the present invention

[0059] 1. Preparation of EB virus Rta protein and polypeptide

[0060] Preparation: inoculate the CHO / RTA cell line with the preservation number: CGMCC6955 into the cell culture dish, the complete culture solution of the cells must be supplemented with 0.1mmol / L hypoxanthine and 0.016mmol / L thymine, 10% fetal bovine serum (Hangzhou Sijiqing Bioengineering Co., Ltd.), after the cells reach a density of about 90%, replace with a serum-free medium without aminomethylpterate (MTX), collect the cell culture medium after 1-2 days, and use trichloro Proteins in cell culture fluid were extracted by acetic acid precipitation.

[0061] Purification: Use GE's streptavidinsepharose highperformance to purify the target protein in the cell supernatant. According to the operation manual of BCA-100 protein quantification kit (Beijing Saichi Biology, product number 300001-B), the test is carried out to quantitatively measure ...

Embodiment 2

[0079] Embodiment 2. Utilize the kit detection sample of the present invention

[0080] Detect 45 parts of nasopharyngeal carcinoma samples and 45 parts of healthy check-up samples with kit of the present invention to check: the steps are as follows:

[0081] (1) Obtain standard curve and curve equation,

[0082] The abscissa is the gradient concentration value of the Rta / IgG calibrator, and the ordinate is the fluorescence value obtained by performing the following steps with the gradient concentration of the Rta / IgG calibrator;

[0083] (2) Add working solution 1 and working solution 2 successively on the slide, incubate and dry;

[0084] (3) Add the sample to be tested diluted by the sample diluent, incubate and dry;

[0085] (4) Add working solution 3, incubate and dry;

[0086] (5) After washing with washing solution, place the glass slide under a fluorescence microscope to observe the reading;

[0087] (6) Substitute the fluorescence value into the standard curve to ...

Embodiment 3

[0101] Embodiment 3. Use the test kit prepared by Rta protein as the reaction antigen to detect the comparative experimental data of the same sample

[0102] Using the kit prepared with Rta protein as the reactive antigen to detect 45 nasopharyngeal carcinoma samples and 45 healthy physical examination samples, the test results are as follows:

[0103] Table 4. Detection results of normal human EB virus Rta-IgG content

[0104]

[0105] Table 5. Detection results of EB virus Rta-IgG content in patients with nasopharyngeal carcinoma

[0106]

[0107]

[0108] It can be known from the experimental results that the sensitivity and specificity of the kit prepared using Rta protein as the reactive antigen are 91.2% and 93.2%, both of which are lower than the sensitivity and specificity of the kit of the present invention.

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Abstract

The invention provides a chemiluminescence kit for detecting EB virus Rta / IgG antibody and a use thereof and belongs to the fields of biotechnology, medical immunology and external serology diagnosis. The chemiluminescence kit comprises magnetic particles coated with an anti-His label monoclonal antibody, a Rta protein with a His label, a specific synthetic polypeptide with a His label and an acridinium ester-labeled anti-human IgG antibody. The specific synthetic polypeptide has an amino acid sequence shown in the formula of SEQ ID NO: 1. The invention also provides a method for detecting an EB virus Rta / IgG antibody by the chemiluminescence kit. The chemiluminescence kit and the detection method can realize fast, accurate, efficient, sensitive and specific detection of an EB virus Rta protein antibody in a sample to be detected, realize multiple sample treatment, shorten detection time and improve detection efficiency.

Description

technical field [0001] The invention relates to the technical fields of biotechnology, medical immunology and in vitro serological diagnosis, in particular to a chemiluminescent reagent kit for detecting Epstein-Barr virus Rta / IgG antibodies and its application. Background technique [0002] Nasopharyngeal carcinoma refers to malignant tumors that occur in the nasopharynx between the nasal cavity and pharynx, accounting for 78.08% of head and neck malignant tumors and 92.99% of upper respiratory tract cancers. Nasopharyngeal carcinoma is most likely to occur in the prime of life between 35 and 50 years old, accounting for 60% of the number of cases. It is easy to cause a major impact on society, economy, labor and family, and is very harmful. [0003] Years of research have found that nasopharyngeal carcinoma is most closely related to Epstein-Barr virus. Epstein-Barr virus is activated to infect nasopharyngeal cells, and when it enters a cleavage and replication state from ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569G01N33/532
CPCG01N33/532G01N33/56994G01N33/68
Inventor 吴凡卢晓希焦守恕李全
Owner 同昕生物技术(北京)有限公司
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