A kind of immunocolloidal gold test paper for rapid detection of mycoplasma bovis and preparation method thereof

The technology of colloidal gold test paper and Mycoplasma bovis is applied in the field of immunological detection, which can solve the problems of inability to realize rapid detection on site, long detection time, complicated operation, etc., and achieves the effects of fast detection speed, low detection cost and improved detection efficiency.

Active Publication Date: 2022-02-11
CHONGQING UNIV OF TECH +1
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Problems solved by technology

[0005] For above-mentioned deficiencies in the prior art, the object of the present invention is to provide a kind of immunocolloidal gold test paper and preparation method thereof of quick detection Mycoplasma bovis, solve the existing Mycoplasma bovis detection method to have complicated operation, long detection time, can not realize on-the-spot rapid detection and high cost

Method used

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  • A kind of immunocolloidal gold test paper for rapid detection of mycoplasma bovis and preparation method thereof
  • A kind of immunocolloidal gold test paper for rapid detection of mycoplasma bovis and preparation method thereof
  • A kind of immunocolloidal gold test paper for rapid detection of mycoplasma bovis and preparation method thereof

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Embodiment

[0047] 1. Antigen screening

[0048] The cell membrane surface protein of Mycoplasma bovis plays an important role in the process of pathogenicity and infection. The P33 gene was screened out by predicting the signal peptide and epitope analysis of the cell membrane surface protein of Mycoplasma bovis, and the bioinformatics of the P33 gene and its encoded protein were further carried out. The results of biological and immunological analysis prove that the P33 protein is a specific adhesion protein on the cell membrane of Mycoplasma bovis.

[0049] 2. Prokaryotic expression and detection of Mycoplasma bovis P33 protein

[0050] 2.1 Construction of pET28a(+)-P33 recombinant vector

[0051] According to the gene sequence of P33 (accession number AIA33909.1) published in GenBank, without changing the amino acid sequence of P33 protein, the gene sequence was optimized according to the codon preference of Escherichia coli, and a pair was designed using Primer Premier5.0 software. ...

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Abstract

The invention discloses an immune colloidal gold test paper for rapid detection of Mycoplasma bovis and a preparation method thereof. The bottom layer of the test paper is a support plate, and closely connected sample pads, bonding pads, NC membranes and absorption pads are sequentially adhered to the support plate , colloidal gold-labeled Mycoplasma bovis P33 monoclonal antibody is adsorbed on the binding pad, rabbit anti-Mycoplasma bovis P33 polyclonal antibody is sprayed on the NC membrane as a detection line and goat anti-BALB / c mouse IgG antibody is sprayed as a quality control line, and the detection The line width of the line and the quality control line is 1mm, the distance between the detection line and the quality control line is 5mm, and they are located in the center of the NC membrane. The amino acid sequence of the Mycoplasma bovis P33 protein is shown in SEQ ID NO.1. The test paper can be used for rapid detection of Mycoplasma bovis antigen, the result can be obtained within 5-10 minutes, and the sensitivity can reach 5ng / mL. It has the advantages of less sample consumption, no cross-reaction and high sensitivity. detection efficiency. It will play an important role in the prevention and control of bovine mycoplasma disease, and has potential application value.

Description

technical field [0001] The invention relates to the technical field of immunological detection, in particular to an immune colloidal gold test paper for rapidly detecting mycoplasma bovis and a preparation method thereof. Background technique [0002] Mycoplasma bovis (Mycoplasma bovis, Mb) is a kind of simple and extremely small prokaryotic microorganisms without cell walls, between bacteria and viruses, capable of self-replication and independent living, which can cause bovine pneumonia, mastitis, arthritis and many other diseases. This disease has caused huge economic losses and seriously threatened the healthy development of my country's cattle industry. [0003] At present, the detection of Mycoplasma bovis in the prior art mainly includes methods such as pathogen isolation and identification, polymerase chain reaction (PCR), fluorescent quantitative PCR technology, enzyme-linked immunosorbent assay (ELISA), but these methods are all subject to clinical application. Lar...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/532G01N33/558G01N33/68
CPCG01N33/532G01N33/558G01N33/68
Inventor 吴胜昔梁望旺曾政李令臣李俊萱侯力嘉鲁友铭蔺露陈忠琼
Owner CHONGQING UNIV OF TECH
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