Colloidal gold test strip for detecting antibodies against akabane virus and preparation method thereof

The technology of colloidal gold test paper and colloidal gold is applied in the field of detection reagents for Akabane disease antibodies, which can solve the problems of complex preparation methods, and achieve the effects of simple preparation methods, fast detection and good specificity.

Inactive Publication Date: 2011-01-19
NINGBO ACAD OF SCI & TECH FOR INSPECTION & QUARANTINE
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Problems solved by technology

[0004] Akabane disease antibody is the immune response protein of Akabane disease virus. Those skilled in the art can obtain monoclonal antibody or polyclonal antibody with conventional purification techniques. For example, the publication number is CN101200709, and the t

Method used

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  • Colloidal gold test strip for detecting antibodies against akabane virus and preparation method thereof
  • Colloidal gold test strip for detecting antibodies against akabane virus and preparation method thereof
  • Colloidal gold test strip for detecting antibodies against akabane virus and preparation method thereof

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[0017] Example

[0018] Preparation process of colloidal gold test strip for detecting Akabane disease antibody

[0019] 1) RNA preparation: Use a viral RNA extraction kit to extract and isolate the envelope glycoprotein G1 gene from the Akabane disease virus. Use the G1 gene as a template to perform RT-PCR amplification reaction and sequence to obtain the G1 gene and the G1 gene. The nucleotide sequence is shown in SEQ ID NO.1 in the sequence table, the nucleotide sequence of the upstream primer of the RT-PCR amplification reaction is shown in SEQ ID NO.3 in the sequence table, and the nucleotide sequence of the downstream primer is shown in the sequence table. SEQ ID NO.4 is shown; the specific RT-PCR amplification reaction is a conventional method and will not be described in detail here; the primers are synthesized by Dalian Baosheng Biological Co., Ltd.

[0020] 2) Preparation of G1 recombinant protein: construct the above-mentioned G1 gene into the expression vector pET-28a(+)...

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Abstract

The invention discloses a colloidal gold test strip for detecting antibodies against akabane virus and a preparation method thereof. The test strip comprises a bottom lining, and an absorbent pad, a nitrocellulose membrane, a glass fiber membrane and a sample mat are arranged on the bottom lining; a detection belt and a quality-control belt are arranged on the nitrocellulose membrane; and G1 recombinant proteins are coated in the detection belt, the antibodies against the akabane virus are coated in the quality-control belt, and colloidal gold-G1 recombinant proteins are coated in the glass fiber membrane, wherein the amino acid sequence of the G1 recombinant protein is shown in a sequence table SEQ ID NO.2. The colloidal gold test strip of the invention has the advantages of simple, efficient, peculiar and sensitive detection, low cost, and the like. The method for preparing the colloidal gold test strip comprises the steps of preparing RNA; preparing the G1 recombinant proteins; preparing the colloidal gold-G1 recombinant proteins; and preparing the antibodies against akabane and preparing the colloidal gold test strip. By using the preparation method, the colloidal gold test strip with good specificity and high sensitivity can be obtained, and meanwhile, the preparation method is simple, and low in cost.

Description

technical field [0001] The invention relates to a detection reagent for Akabane disease antibody, in particular to a colloidal gold test strip for detecting Akabane disease antibody and a preparation method thereof. Background technique [0002] Akabane disease, also known as Akabane disease, is caused by Akabane virus (AKAV), which can lead to miscarriage, premature birth, stillbirth and birth of congenital arthrogryposis-hydrocephalus anencephaly syndrome fetus. It is the four major abortions in cattle my country's Ministry of Agriculture has also listed this disease as an important infectious disease that is prohibited from entering and leaving the country. The disease is distributed in tropical and temperate regions. In 2094, Li Changlin et al. tested 2639 serums of cattle and sheep across the country. The positive rate of cattle was 39.18%, and that of sheep was 12.6%. The positive rate in the south was higher than that in the north. In 1999, Wei Longxing and others con...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/558G01N33/531C12N15/40C12N15/70C07K16/10C07K16/06
Inventor 张吉红闻伟刚吴东来李如松黄素文
Owner NINGBO ACAD OF SCI & TECH FOR INSPECTION & QUARANTINE
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