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Haemophilus parasuis test paper strip and preparation method thereof

A technology of Haemophilus suis and test strips, which is applied in measurement devices, instruments, scientific instruments, etc., can solve the problems of high antigen requirements, complicated preparation, affecting field epidemiology, etc., and achieves strong sensitivity and good specificity. Effect

Active Publication Date: 2014-12-31
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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Problems solved by technology

[0002] Due to the large number of serotypes of Haemophilus parasuis, 15 serotypes are currently known, and there are many methods for detecting the pathogen and antibodies, but they are all species-specific. The detection of Haemophilus parasuis disease Methods include PCR method for detecting antigens, indirect hemagglutination method and ELISA method for detecting serum antibodies. Only the agar amplification test developed by the applicant can be used for stereotype detection. The preparation of agar expansion test is complicated, and it takes 2-3 days for the detection to produce results. In addition, the tested serum needs to be concentrated, and the requirements for antigens are high
Haemophilus parasuis type 5 is one of the popular strains of Haemophilus parasuis disease in my country at present, but the existing technology lacks antibodies capable of typing and detecting type 5 serotypes, especially rapid detection methods, thus affecting field epidemics Scientific investigation and accurate prevention and control work

Method used

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  • Haemophilus parasuis test paper strip and preparation method thereof
  • Haemophilus parasuis test paper strip and preparation method thereof
  • Haemophilus parasuis test paper strip and preparation method thereof

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Embodiment Construction

[0020] The present invention is illustrated below in conjunction with examples.

[0021] The antigen coated on the gold standard pad used in the test strip of the present invention is Haemophilus parasuis OPPA fusion protein. The antigen coated by the gold standard pad of the present invention is Haemophilus parasuis OPPA fusion protein, which is a specific protein shared by 15 serotypes of Haemophilus parasuis. The sequence was obtained in vitro, and the sequence was optimized. The fusion tag was obtained, and then the protein was cloned and expressed. After expression, it went through a series of purifications, and then applied to the test strip, the purpose of which was to improve the specificity of the detection test strip. Its preparation method is as follows:

[0022] 1 Primer design and selection of enzyme cutting sites

[0023] According to the HPS OppA gene sequence in GenBank (SH0165 strain accession number: NC_011852.1), a pair of specific primers were designed us...

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Abstract

The invention discloses a haemophilus parasuis type 5 colloidal gold formed kit and a preparation method thereof. The test paper strip comprises a sample pad, a colloidal gold pad, a nitrocellulose membrane and an absorbent pad which are orderly connected and fixed on a PVC (polyvinyl chloride) plate, and also comprises a detection line and a quality control line, the sample pad comprises a water absorbent material, the colloidal gold pad is coated with antigen, the detection line and the quality control line are located on the nitrocellulose membrane and are separated from each other, the antigen coating the colloidal gold pad is haemophilus parasuis OPPA (oligopeptide binding protein A) fusion protein, the detection line is haemophilus parasuis type 5 antigen, and the quality control line is haemophilus parasuis whole bacterial antigen polyclonal antibody IgG. The test paper strip has the advantages of good specificity, strong sensitivity and repeatability.

Description

technical field [0001] The invention relates to a disease detection reagent used in animal husbandry and a preparation method thereof, in particular to a Haemophilus parasuis type 5 colloidal gold typing kit and a preparation method of the kit. Background technique [0002] Due to the large number of serotypes of Haemophilus parasuis, 15 serotypes are currently known, and there are many methods for detecting the pathogen and antibodies, but they are all species-specific. The detection of Haemophilus parasuis disease Methods include PCR method for detecting antigens, indirect hemagglutination method and ELISA method for detecting serum antibodies. Only the agar amplification test developed by the applicant can be used for stereotype detection. The preparation of agar expansion test is complicated, and it takes 2-3 days for the detection to produce results. In addition, the serum to be tested needs to be concentrated, and the requirements for antigens are high. Haemophilus pa...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/531
CPCG01N33/531G01N33/569
Inventor 贺英逯忠新赵萍储岳峰陈胜利张建军
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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