ELISA kit for detecting IgG antibody of feline calicivirus and detecting method thereof

A feline calicivirus and kit technology, which is applied to the ELISA kit for detecting feline calicivirus IgG antibody and its detection field, can solve problems such as delayed treatment, time-consuming detection, virus diffusion and transmission, and achieve detection The conclusion is clear and clear, the operation time is short, and the effect of high repetition stability

Inactive Publication Date: 2016-10-26
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] Diagnosis based on medical history, clinical symptoms and epidemic characteristics is often misdiagnosed, delays treatment and leads to the spread and spread of the virus
At present, the detection of feline calicivirus antibody uses methods such as serum neutralization test and indirect immunofluorescence, which are time-consuming, poor in repeatability, and mostly used in laboratories, lacking uniform standards

Method used

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  • ELISA kit for detecting IgG antibody of feline calicivirus and detecting method thereof
  • ELISA kit for detecting IgG antibody of feline calicivirus and detecting method thereof
  • ELISA kit for detecting IgG antibody of feline calicivirus and detecting method thereof

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specific Embodiment approach 1

[0028] Specific embodiment one: the ELISA kit that this embodiment is used to detect feline calicivirus IgG antibody comprises ELISA microwell plate, enzyme-labeled secondary antibody and substrate chromogenic solution; Wherein, ELISA microwell plate contains coated Soluble expression of feline calicivirus ORF2 recombinant protein F.

[0029] The preparation method of the FCV gene ORF2 recombinant protein F of soluble expression in the present embodiment:

[0030] According to the feline calicivirus (FCV) VP1 gene sequence (Genbank: X99445.1) registered in Genbank, the primers for amplifying the F region RNA fragment of feline calicivirus ORF2 were designed, and the PCR method was used to amplify, and then the amplified The gene fragment was cloned into the vector pET-32a to construct a recombinant plasmid, and after the sequence verification was correct, it was transferred into the expression host E.coli BL21 (DE3) to induce expression with IPTG; then optimize the induction c...

specific Embodiment approach 2

[0034] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the ELISA kit for detecting IgG antibodies to feline calicivirus also includes a stop solution, which is a sulfuric acid solution. Other steps and parameters are the same as those in Embodiment 1.

[0035] In this embodiment, the stop solution is a sulfuric acid solution with a concentration of 2 mol / L.

specific Embodiment approach 3

[0036]Embodiment 3: The difference between this embodiment and Embodiment 1 or 2 is that the substrate chromogenic solution is TMB chromogenic solution. Other steps and parameters are the same as those in Embodiment 1 or 2.

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Abstract

The invention relates to a kit for detecting feline calicivirus and a detecting method thereof, in particular to an ELISA kit for detecting an IgG antibody of feline calicivirus and a detecting method thereof. The ELISA kit and the detecting method solve the problem that current feline calicivirus diagnosis and detection results are not ideal. The kit comprises an ELISA microwell plate, RaMIgG-HRP and substrate developing liquid. The method comprises the steps that firstly, serum is diluted and added into the ELISA microwell plate to be incubated; secondly, the RaMIgG-HRP is added for incubation; thirdly, the substrate developing liquid is added, and reaction is carried out in a shading mode at the room temperature; fourthly, a stop solution is added to terminate the reaction, an ELIASA is used for detecting 450 nm light absorption value, the 450 nm light absorption value and a Cut Off value of the ELISA kit are compared, and then a detection result is obtained. The ELISA kit detection result is high in repeatability and stability, and the speed is high.

Description

technical field [0001] The invention relates to a kit for detecting feline calicivirus and a detection method thereof. Background technique [0002] Feline calicivirus (Feline calicivirus, FCV) is a single-stranded RNA virus belonging to the family Caliciviridae and belonging to the genus Lysovirus. FCV is a viral respiratory infectious disease in cats. The main manifestations are oral ulcers, increased eye and nose secretions, salivation, conjunctivitis, shame, stomatitis, tracheitis, and bronchitis, accompanied by biphasic fever. It is a frequently-occurring disease in cats with a high incidence rate. Infection with virulent strains can cause pneumonia, dyspnea, and even death. The main sources of infection are sick cats and virus-infected cats. In the acute stage, sick cats can excrete a large amount of virus with secretions and excretions, directly infecting susceptible cats; after treatment, the symptoms of virus-infected cats can disappear, and they still shed the vir...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56983G01N2333/145
Inventor 刘家森曲连东田进刘大飞刘春国郭东春李志杰刘明姜骞胡晓亮康洪涛吴红霞
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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