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Avian H9 subtype avian influenza virus antibody colloidal gold test paper and test card

A technology of colloidal gold and avian influenza, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of not being able to target a variety of poultry at the same time, and cannot dynamically and quantitatively monitor the antibody level of poultry groups, so as to increase objectivity and detection time Short, the effect of reducing the error rate

Inactive Publication Date: 2019-07-26
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In the process of realizing the present invention, the inventors have found that there are at least the following problems in the prior art: the above-mentioned colloidal gold detection test paper can only perform qualitative or semi-quantitative detection on chicken-related diseases or antibody levels after vaccine immunization, but cannot simultaneously target multiple Poultry, and cannot dynamically and quantitatively monitor the antibody level of the poultry population

Method used

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  • Avian H9 subtype avian influenza virus antibody colloidal gold test paper and test card
  • Avian H9 subtype avian influenza virus antibody colloidal gold test paper and test card
  • Avian H9 subtype avian influenza virus antibody colloidal gold test paper and test card

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Experimental program
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Effect test

Embodiment 1

[0086] 1. AIV-HA1 9 Preparation of recombinant protein

[0087] 1. Cloning and sequencing of 1H9 subtype avian influenza HA1 gene and recombinant plasmid pGEX-KG-HA1 9 build

[0088] The sequence and cloning method and expression vector pGEX-KG-HA1 of the H9 subtype avian influenza HA1 gene involved in the present invention 9 The reference for the construction method is: Lei Ting, Cloning and expression of H9 subtype avian influenza virus HA1 gene and preliminary analysis of its biological activity [J]. Chinese Journal of Preventive Veterinary Medicine, 2006. The specific implementation method is as follows: use the AIV-H9N2 virus that is freeze-dried and preserved in the Microbiology and Immunity Laboratory of Huazhong Agricultural University, inoculate AIV-H9N2 in the allantoic cavity in 9-11 days old healthy SPF (specific pathogen-free) chicken embryos (purchased from Wuhan Institute of Biological Products), discard dead embryos 24 hours ago, collect chicken embryo allan...

Embodiment 2

[0141] Evaluation of the stability of the colloidal gold detection test paper for the avian H9 subtype avian influenza antibody of the present invention

[0142] The vacuum-packaged test papers of the present invention placed at 4°C and 37°C respectively were taken out on the 7th, 16th, 35th, 54th, 70th, and 90th day, and the H9 subtype avian influenza standard positive serum was diluted with pure water in multiples. detection. The results are shown in Table 4. The test paper of the present invention has a longer shelf life at 4° C., and the shelf life is at least 90 days.

[0143] The shelf life test result under the different storage conditions of table 4 test paper

[0144]

[0145] Note: "+" indicates that the detection line develops color, and "-" indicates that the detection line does not develop color.

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Abstract

The invention provides avian H9 subtype avian influenza virus antibody colloidal gold test paper and test card. The avian H9 subtype avian influenza virus antibody colloidal gold test paper and test card can detect the immune level of various birds and are wide in application range, high in specificity, high in sensitivity, easy to operate and short in consumed time. The test paper comprises a sample pad, a gold-labeled pad and a nitrocellulose membrane which are sequentially connected, wherein the gold-labeled pad is wrapped with a colloidal gold label of a monoclonal antibody, namely anti-mouse IgY Fc CH3-CH4; and a detection line close to one end of the gold-labeled pad and a quality control line away from one end of the gold-labeled pad are arranged on the nitrocellulose membrane, thedetection line is wrapped with an AIV-HA19 recombinant protein, the quality control line is wrapped with a second antibody, and the DNA sequence for encoding the AIV-HA19 recombinant protein is shownas SEQ ID NO.1.

Description

technical field [0001] The invention relates to the technical field of animal immunity application, in particular to a colloidal gold detection test paper and a test card for poultry H9 subtype avian influenza antibody. Background technique [0002] H9 subtype avian influenza virus (AIV) exists in poultry flocks for a long time, causing huge economic losses to the poultry industry. Since the first isolation in 1994, the H9N2 subtype avian influenza virus (H9N2AIV) has been widely distributed in my country and is one of the common pathogens in poultry industry in my country. In 1994, the H9N2 avian influenza virus was isolated for the first time in a chicken farm in Guangdong, my country. The clinical manifestations were a decrease in egg production rate and a certain mortality rate. After the occurrence of this AIV, H9N2AIV has been on the rise in my country, seriously restricting the development of the poultry industry. Waterfowl are known as the "natural gene repository"...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/531G01N33/543
CPCG01N33/531G01N33/54306G01N33/56983
Inventor 李自力邵雨胡思顺毕丁仁王喜亮肖运才金卉
Owner HUAZHONG AGRI UNIV
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