Portable lactating mastitis pathogenic bacteria MRSA detection method

A detection method and lactation technology, applied in the field of medical detection, can solve the problems of activity, affinity and titer reduction, and achieve the effects of reducing medical disputes, strengthening transformation ability, and reducing economic burden

Inactive Publication Date: 2020-01-07
WUHAN UNIV
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Problems solved by technology

[0013] 4. Insufficient
[0014] The decrease in activity, affinity and titer of antibodies after coupling to magnetic beads will often affect the effect of downstream technology applications and become a key factor restricting the application of this enrichment technology

Method used

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  • Portable lactating mastitis pathogenic bacteria MRSA detection method
  • Portable lactating mastitis pathogenic bacteria MRSA detection method
  • Portable lactating mastitis pathogenic bacteria MRSA detection method

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Embodiment Construction

[0046] The present invention will be further described in detail below in conjunction with specific embodiments and accompanying drawings.

[0047] (1) The magnetic nanoparticles used in the embodiments of the present invention are magnetic nanoparticles with a particle size of 800nm ​​modified by the base, purchased from Wuhan Jiayuan Quantum Dot Technology Development Co., Ltd. PBP2a aptamer, by Sangon Bioengineering (Shanghai ) Co., Ltd., address: No. 698, Xiangmin Road, Songjiang District, Shanghai, deposit number: M23.

[0048] (2) the structure of the immune colloidal gold immunochromatography test paper used in the embodiment of the present invention, see figure 2 , the reaction support is a 6.2cmx0.4cmPCV plate; the absorbent pad is 2cmx0.4cm oil filter paper; the nitrocellulose membrane of 1.8cmx0.4cm is coated with mouse anti-PBP2a protein (Qac protein) antibody (purchased from Dingguo Biotechnology Company, 2mg / ml), goat anti-mouse IgG1mg / ml (containing 1% BSA blo...

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Abstract

The present invention discloses a portable lactating mastitis pathogenic bacteria MRSA (Methicillin-resistant Staphylococcus aureus) detection method. Rapid detection of the MRSA is performed throughthe combined use of a nano-immunomagnetic bead technology and a colloidal gold immunochromatographic technology. The method specifically comprises the following steps that: nano-magnetic beads are coupled with a monoclonal antibody obtained by using the hybridoma technology to form nano-immunomagnetic beads; the other paired monoclonal antibody labeled with the colloidal gold is taken as a labeledantibody, and the polyclonal serum and the goat anti-mouse IgG are used as the detection line T and the quality control line C, respectively; a binding pad containing the colloidal gold-labeled antibody, a chromatography membrane containing the detection line T and the quality control line C, and a water-absorbing pad constitute a colloidal gold immunochromatographic test paper; and when detecting a sample, the immunomagnetic beads and the bacteria in the sample are specifically captured and enriched, the enriched liquid is added to the lysate for lysis, then the lysate is dropped on the sample pad of the test paper, and a visible coloration band is formed at the detection line T and the quality control line C according to the color of the colloidal gold, so that rapid and semi-quantitative detection of the MRSA is realized.

Description

technical field [0001] The invention belongs to the technical field of medical detection, and specifically refers to a portable method for detecting MRSA, a pathogenic bacterium of lactating mastitis. Background technique [0002] MRSA is a stubborn pathogen of lactating mastitis. The purpose of this product is to diagnose and treat early, improve the cure rate of the disease, and reduce the damage to the breast during the treatment. At present, there is no portable technology product for rapid detection of MRSA in breast milk clinically. [0003] The immunomagnetic bead enrichment technology uses magnetic microspheres as the solid phase surface, combined with immunological methods to establish a sample concentration method with important application prospects. Magnetic beads form immunomagnetic beads by binding antibodies, which can specifically bind to corresponding antigens in the liquid phase, and rely on the force of the magnetic field to rapidly concentrate the requir...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/543G01N33/532
CPCG01N33/532G01N33/54306G01N33/54326G01N33/54346G01N33/56938
Inventor 陈创刘辉凡吴淑娟谭细容刘长江张智辉
Owner WUHAN UNIV
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