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EⅢ-indirect elisa antibody detection kit and application for detecting porcine Japanese encephalitis virus

A swine Japanese encephalitis and antibody detection technology, applied in the biological field, can solve the problems of many influencing factors, time-consuming and labor-intensive, etc., and achieve a highly specific effect

Active Publication Date: 2015-09-30
广州易安生物技术有限公司
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  • Summary
  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

The isolation and identification of pathogens is the most traditional detection method, and its results are accurate and reliable, but there are many influencing factors, and the actual operation is quite time-consuming and laborious, so it has certain limitations in clinical application; RT-PCR technology requires special equipment ( Such as PCR instrument and gel imaging system, etc.) and professionals to carry out relevant operations; enzyme immunoassay is one of the most commonly used methods in serological methods, and enzyme immunoassay combines the amplification effect of enzyme-catalyzed reaction with antigen-antibody affinity Combined with the high specificity and specificity of the reaction, the immunoassay with enzyme-labeled antigen or antibody as the main reagent has high sensitivity and specificity. However, the sensitivity and specificity of the immunoassay depend on the specificity of the antigen. choose

Method used

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  • EⅢ-indirect elisa antibody detection kit and application for detecting porcine Japanese encephalitis virus
  • EⅢ-indirect elisa antibody detection kit and application for detecting porcine Japanese encephalitis virus
  • EⅢ-indirect elisa antibody detection kit and application for detecting porcine Japanese encephalitis virus

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Embodiment 1

[0058] I. Materials

[0059] KOD enzyme used in the following examples is TOYOBO company product;

[0060] The JEV-EⅢ protein is a protein expressed and purified in BL21 (DE3) Escherichia coli at a concentration of 4.7 μg / mL;

[0061] The enzyme plate is the product of JET company;

[0062] The washing solution was prepared by the following steps: 3.579g Na 2 HPO 4 12H 2 O, 1.56gNaH 2 PO 4 2H 2 O, 29.215g NaCl and 600mL ddH 2 O is fully dissolved, add 0.5mL Tween-20, adjust the pH value to 7.4 with 10mol / L HCl solution, ddH 2 O was fixed to 1000mL;

[0063] Coating buffer (pH9.60.05mol / L carbonate buffer): 1.5g sodium carbonate, 2.93g sodium bicarbonate dissolved in 600mL ddH 2 O, adjust pH to 9.6, ddH 2 O was fixed to 1000mL;

[0064] Serum diluent is soybean lecithin solution with a mass volume ratio (g / mL) of 0.5%, and the solvent is washing liquid;

[0065] The blocking solution is a soybean lecithin solution with a mass volume ratio (g / mL) of 0.5%, and the so...

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Abstract

The invention discloses an EIII-indirect ELISA (Enzyme-Linked Immuno Sorbent Assay) antibody detection kit for detecting a swine Japanese encephalitis virus and an application thereof. The kit disclosed by the invention consists of an elisa plate which takes a purified JEV-EIII protein as an envelope antigen, a cleaning liquid, a serum diluent, rabbit anti-pig elisa secondary antibody, a primer developing liquid, a stop buffer, a JEV positive serum and a JEV negative serum. The envelope antigen used by the invention is easy to be obtained stably and massively, the purifying method is simple and easy to realize, and concentration of recombinant proteins is easy to test and control, thereby facilitating industrialized production on a large scale. The kit disclosed by the invention is used for detecting the swine Japanese encephalitis virus antibody, and the detection coincidence rate with the ELISA kit in the prior art is 90%. The kit disclosed by the invention is convenient to operate, high in sensitivity, good in specificity, low in using cost, good in repeatability and suitable for popularization and application, and provides a reliable means for clinical rapid detection of the JEV antibody.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to an EⅢ-indirect ELISA antibody detection kit for detecting porcine Japanese encephalitis virus and its application. Background technique [0002] Japanese encephalitis virus (JEV) belongs to the Flavivirus genus of the family Flaviviridae, and the Japanese encephalitis caused by it is a central nervous system zoonotic acute infectious disease. It is estimated that about 50,000 people in the world are infected with JEV every year, about 15,000 cases die, and the mortality rate is about 30%. At the same time, JEV is one of the important viruses that seriously harm the pig industry, causing huge economic losses to the pig industry The export of meat products is restricted. In 2008, the Ministry of Agriculture of my country classified swine Japanese encephalitis disease as a second-class animal disease. [0003] The disease was first reported in Japan in the 1870s, and Japan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/09
CPCG01N33/531G01N33/56983G01N2333/185
Inventor 陈金顶姚俊庸郑仲华邓洁汝刘翠翠勾红潮裴晶晶
Owner 广州易安生物技术有限公司
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