Triple rapid detection kit of three avian respiratory diseases, and application thereof

A test card, avian influenza virus technology, applied in measuring devices, antiviral immunoglobulins, instruments, etc., can solve the problems of poor case diagnosis, waste of time and resources, and inability to accurately reflect whether chickens are infected with epidemic diseases. , to achieve the effect of short detection time, intuitive judgment and high sensitivity

Active Publication Date: 2014-08-27
HUAZHONG AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the above kits are for the detection of antibody levels after immunization of related diseases or vaccines, and cannot accurately reflect whether the chickens are infected with the disease
Although there are also some immunocolloidal gold detection methods for a single pathogen at present, such as the "Avian Influenza Virus Antigen Detection Method and Its Gold Standard Rapid Diagnosis Kit and Preparation Method" with the patent application number 200510042631. "Influenza immune colloidal gold diagnostic test strips and test cards" discloses several preparation methods for detecting avian influenza virus antigens, but the diagnostic effect on mixed infection cases is not good, and multiple single-detection colloidal gold test strips are used for detection waste of time and resources

Method used

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  • Triple rapid detection kit of three avian respiratory diseases, and application thereof
  • Triple rapid detection kit of three avian respiratory diseases, and application thereof
  • Triple rapid detection kit of three avian respiratory diseases, and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] 1. Preparation of AIV-HA1 protein, NDV-HN protein and IBV-NP protein

[0048] The preparation methods of AIV-HA1 protein, NDV-HN protein and IBV-NP protein mainly refer to the literature: Sambrook J, Fritsch E F, Manny Artis T (translated by Jin Dongyan, Li Mengfeng, etc.). Molecular Cloning Experiment Guide .Second Edition, Science Press, 1992 provided the method.

[0049] 1.1 Cloning and sequencing of avian influenza virus HA1 gene and construction of expression vector pKG-NS1

[0050] The construction method and construction map of the avian influenza virus HA1 gene and its recombinant expression plasmid pKG-HA1 involved in the present invention have been published on CNKI in 2006, refer to the literature: CNKI Dissertation Library: Wu Renwei, Recombinant Protein of Avian Influenza Virus Mucosal immunity research and establishment of N-ELISA antibody detection method [D] Huazhong Agricultural University, 2006; see website: http: / / epub3.cnki.net / KCMS / detail / detail.as...

Embodiment 2

[0089] Embodiment 2 (application embodiment)

[0090] Method of using triple rapid detection kit for poultry major respiratory diseases

[0091] 1. The composition of the kit, including:

[0092] ① 25 test cards

[0093] ②One bottle of sample diluent (10mL / bottle)

[0094] 2. Sample Preparation

[0095] 2.1 Sample diluent: The sample diluent is 0.85% sodium chloride solution. Preparation method: 8.5g NaCl, add distilled water to make up to

[0096] 1000mL.

[0097] 2.2 Sample preparation

[0098] The detailed operation method of sample collection and processing is as follows:

[0099] (1) Sample of trachea and pharynx: First, remove the chicken's howl by hand, then pull out the chicken's tongue by hand or tweezers to expose the pharynx, insert a cotton swab into the trachea of ​​the pharynx, stir it a few times, take it out, and then Put it into the sample tube that has been added with 500 μL of sample diluent in advance, stir and squeeze vigorously to elute the sample...

Embodiment 3

[0105] 1 Example of the application of the triple rapid detection kit for major poultry respiratory diseases

[0106] 1.1 Detection of Chicken Standard Antigen

[0107] ①Specificity test: Commercially purchased avian influenza H5 subtype HI antigen, avian influenza H9 subtype HI antigen, chicken Newcastle disease HI antigen, infectious bronchitis HI antigen (the above biological materials were purchased from Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences) , Egg Drop Syndrome (EDSV-76) HI Antigen, Infectious Bursal of Fabricius (IBD) Agar-Extended Antigen, Mycoplasma Gallisepticum Antigen, Chicken Infectious Laryngotracheitis (AILT) Virus Antigen (the above biological materials were purchased from Beijing . China Veterinary Drug Administration), 0.85% sodium chloride solution and negative allantoic fluid were tested according to the method described in Example 2 of the present invention (GICA). When the detection test card in the kit of the pres...

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Abstract

The invention belongs to the field of infectious disease detection of animals, and concretely relates to a triple rapid detection kit of three avian respiratory diseases, and an application thereof. The kit comprises a test paper strip, a test card shell and a sample dilution, and the test paper strip is formed by sequentially pasting an absorbing pad, a cellulose nitrate membrane, a colloidal-gold pad and a sample pad on a non-absorbent support slice; a combination pad is coated with an anti-avian influenza virus monoclonal antibody-colloidal gold marker, an anti-Newcastle disease virus monoclonal antibody-colloidal gold marker and an anti-chicken infectious bronchitis virus monoclonal antibody-colloidal gold marker mixture, detection lines T1, T2 and T3 are coated with the anti-avian influenza monoclonal antibody, the anti-Newcastle disease virus monoclonal antibody and the chicken infectious bronchitis monoclonal antibody, and the cellulose nitrate membrane is coated with a rabbit anti-rat IgG quality control line. The kit can simultaneously complete the disease cause detection of bird flu H5 type, H9 subtype, Newcastle diseases and chicken infectious bronchitis, and has strong specificity and high sensitivity.

Description

technical field [0001] The invention belongs to the technical field of animal immunity application. It specifically relates to a triple rapid detection kit and its application for the detection of pathogens of three poultry respiratory diseases (avian influenza H5 subtype and H9 subtype, Newcastle disease and chicken infectious bronchitis). The kit shell prepared by the invention simultaneously completes rapid detection of three poultry respiratory diseases such as avian influenza H5 subtype and H9 subtype, Newcastle disease and chicken infectious bronchitis pathogens at one time. Background technique [0002] Poultry respiratory infection is a very common disease in poultry production practice, especially the occurrence of this kind of disease is complex and changeable, which has brought huge economic losses to the poultry industry. The pathogens that cause respiratory infections in poultry mainly include viruses, bacteria and mycoplasma, among which are three diseases cau...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/571
CPCC07K16/10C07K16/1018C07K16/1027C07K2317/14G01N33/56983G01N2333/08G01N2333/11G01N2333/115
Inventor 肖运才王喜亮石德时刘锡玲苏金玲都跃毕丁仁周祖涛李自力刘梅
Owner HUAZHONG AGRI UNIV
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