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Monoclonal antibody blocking enzyme-linked immunosorbent assay (ELISA) kit and method for detecting nonstructural protein (NSP) antibody of foot-and-mouth disease virus (FMDV)

A non-structural protein, foot-and-mouth disease virus technology, applied in the field of monoclonal antibody blocking ELISA kit, can solve the problem that 3D protein cannot distinguish between infected animals and vaccine immunized animals.

Active Publication Date: 2015-05-27
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Among several non-structural proteins of FMDV, 3D was the first non-structural protein used for diagnosis, but 3D antibodies were also detected from vaccine-immunized animals later, indicating that 3D proteins cannot distinguish infected animals from vaccine-immunized animals

Method used

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  • Monoclonal antibody blocking enzyme-linked immunosorbent assay (ELISA) kit and method for detecting nonstructural protein (NSP) antibody of foot-and-mouth disease virus (FMDV)
  • Monoclonal antibody blocking enzyme-linked immunosorbent assay (ELISA) kit and method for detecting nonstructural protein (NSP) antibody of foot-and-mouth disease virus (FMDV)
  • Monoclonal antibody blocking enzyme-linked immunosorbent assay (ELISA) kit and method for detecting nonstructural protein (NSP) antibody of foot-and-mouth disease virus (FMDV)

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Effect test

Embodiment 2

[0105] Kit composition

[0106] The kit consists of 8 reagents including: enzyme-labeled reaction plate, serum dilution, 25-fold concentrated washing solution, substrate solution, 100× concentrated enzyme-labeled detection antibody, stop solution, positive control serum and negative control serum, and 4 Sheet of sealing film and instructions.

[0107] Preparation method:

[0108] 1. Antigen Coated Plates: Two 96-well high-affinity ELISA plates, first coated with 6×histidine monoclonal antibody or monospecific 2C polyclonal antibody, and passed through monoclonal antibody or 2C polyclonal antibody captures the prokaryotic expression of the histidine-labeled foot-and-mouth disease virus 3ABC or 2C3AB non-structural protein, and the enzyme-labeled plate is vacuum-sealed and stored after the treatment process such as adding a blocking stabilizer and drying. The shelf life can reach 1.5 years.

[0109] 2. Serum Dilution Buffer: a 0.01M phosphate (PBS) solution containing 10% horse serum (...

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Abstract

The invention discloses a monoclonal antibody blocking enzyme-linked immunosorbent assay (ELISA) kit and a method for detecting the nonstructural protein (NSP) antibody of a foot-and-mouth disease virus (FMDV) (FMD NSP B-ELISA); the kit comprises ELISA reaction plates, serum diluent, 25 times concentrated detergent, substrate solution, 100* concentrated ELISA detecting antibody, stop buffer, positive control serum and negative control serum; the ELISA reaction plates are two 96-pore high-affinity ELISA reaction plates, firstly 6* groups of amino acid monoclonal antibody or NSP 2C polyclonal antibody, and then FMDV 3ABC or 2C3AB NSP which is expressed by pronucleus and is provided with 6* groups of amino acid labels is captured through the monoclonal antibody or the polyclonal antibody; and compared with other similar kits, the method has higher coincidence rate and higher positive serum detection rate, and is applicable to detecting the serum of cattle, sheep, pigs and other susceptible animals.

Description

Technical field [0001] The invention relates to a monoclonal antibody blocking ELISA (FMDVNSP B-ELISA) kit and method for detecting foot-and-mouth disease virus non-structural protein antibodies, and belongs to the field of biotechnology. Background technique [0002] Foot-and-mouth disease (FMD) is a severe infectious disease co-infected by cloven-hoofed animals caused by foot-and-mouth disease virus (FMDV) infection. Because of its extremely high incidence, which seriously affects animal husbandry production and international trade, it is highly valued by all countries. Foot-and-mouth disease prevention and control mainly adopt comprehensive prevention and control measures of vaccine immunization and slaughter infection and suspicious animals. Vaccine-immunized animals and infected animals will produce structural protein antibodies. Therefore, the diagnostic method for detecting structural protein antibodies is difficult to distinguish between vaccine-immunized and infected ani...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
Inventor 卢曾军刘在新曹轶梅付元芳孙普李冬包慧芳李平花白兴文陈应理谢宝霞厍大亮
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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