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Feline calicivirus fluorescent quantitative detection card and detection method

A fluorescent quantitative detection, feline calicivirus technology, applied in fluorescence/phosphorescence, measuring device, material analysis by optical means, etc. Wide sample source, easy operation, simple operation effect

Pending Publication Date: 2019-07-09
江苏雷森生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

These detection methods require professional knowledge and professional instruments and equipment, and have high requirements for testing personnel, and the obtained results need to be sorted out, which is time-consuming and labor-intensive

Method used

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  • Feline calicivirus fluorescent quantitative detection card and detection method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The preparation method of embodiment 1. fluorescent latex microspheres, comprises the following steps:

[0034] 1. Activation of fluorescent latex microspheres

[0035] (A) Add a quantitative fluorescent latex microsphere solution to the phosphate buffer solution, mix well and centrifuge at 16000rpm for 20min, then remove the supernatant, add phosphate buffer solution, and ultrasonicate for 60s to obtain fluorescent latex microsphere dispersion liquid.

[0036] (B) Quickly add EDC&NHS activator to the fluorescent latex microsphere dispersion in step (A), mix evenly on a vortex shaker, and shake and activate at room temperature for 30 minutes.

[0037] 2. Coupling of fluorescent latex microspheres

[0038] (C) Centrifuge the fluorescent latex microsphere dispersion liquid activated in step (B) at 16000rpm for 20min, remove the supernatant, wash with the buffer solution in step (A), centrifuge again under the same conditions and discard Supernatant, reconstitute and re...

Embodiment 2

[0046] Embodiment 2. a kind of preparation method of feline calicivirus fluorescent quantitative detection card, comprises the following steps:

[0047] (1) Soak the bonding pad in the phosphate treatment solution in which BSA is dissolved, and after drying, spray the prepared fluorescent latex microsphere dispersion evenly on the bonding pad, and dry in an oven.

[0048] (2) Spray the anti-feline calicivirus monoclonal antibody diluted to 2 mg / mL on the detection line of the reaction membrane at a speed of 1.6 μL / cm, and dry it.

[0049] (3) Spray the goat anti-mouse polyclonal antibody diluted to 2.6 mg / mL on the quality control line of the reaction membrane at a speed of 1.2 μL / cm, and dry it.

[0050] (4) Cut off the bonding pad sprayed with fluorescent latex microspheres, and paste and cover the 2-3 mm of the front end on the reaction film.

[0051] (5) Soak the sample pad in the phosphate treatment solution, dry it, and cover the rear end of the reaction membrane with a...

Embodiment 3

[0055] Embodiment 3. A method for detecting feline calicivirus, comprising the following steps:

[0056] (1) Use a swab to pick up the feces, nasal cavity, oral cavity, eye secretions or serum of the animal to be tested, and dissolve the pick-up in the diluent;

[0057](2) Add the diluent in step (1) to the sample tank, make it gradually infiltrate the sample pad and the binding pad, and finally infiltrate the fluorescent latex microspheres coupled with the anti-feline calicivirus monoclonal antibody into the reaction chamber. Membrane, so that the detection line and quality control line of the reaction membrane can capture fluorescent microspheres;

[0058] (3) Insert the detection card into the supporting quantitative detection instrument, the instrument automatically scans the reaction film under the scanning window, and finally obtains the specific content of feline calicivirus in the diluent;

[0059] (4) Analyze the test results, generate a test report, judge the virus-...

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Abstract

The invention discloses a feline calicivirus fluorescent quantitative detection card and detection method. The fluorescent quantitative detection card is composed of a sample adding groove, a scanningwindow and a handheld device; the sample adding groove covers a sample pad, a bonding pad and part of a reaction membrane; the scanning window covers the reaction membrane; the bonding pad is coatedwith a fluorescent latex microsphere-coupled anti-feline calicivirus monoclonal antibody; the reaction membrane contains a detection line and a quality control line; the detection line is coated withthe anti-feline calicivirus monoclonal antibody; and the quality control line is coated with a goat anti-mouse polyclonal antibody. A quantitative detection result can be provided. In addition, the invention discloses a feline calicivirus disease condition and prognosis judging method based on the quantitative detection result. The method can be used for quantitative detection of feline calicivirus, has the advantages of being high in sensitivity, easy to operate, high in stability and the like, and can realize tracking and monitoring of clinical samples.

Description

technical field [0001] The invention relates to a cat calicivirus fluorescent quantitative detection card and a detection method, belonging to the field of biological detection. Background technique [0002] Feline calicivirus (FCV) belongs to the family Caliciviridae and belongs to the genus Herpesvirus. It is a single-stranded RNA virus with icosahedral symmetry. It can cause oral and respiratory infectious diseases of cats and feline animals, and can also cause conjunctivitis. Feline calicivirus is also named feline infectious rhino-conjunctivitis virus. After infection with the virus, sick animals may develop symptoms such as oral ulcers, increased secretions from the eyes and nose, shame, tracheitis, and bronchitis. Fastier isolated the virus for the first time in 1957, and domestic scientists have also isolated the virus from sick cats since then. The full-length FCV genome is 7690bp, encoding three open reading frames (ORFs), ORF1 encodes a non-structural protein, O...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N21/64
CPCG01N33/56983G01N21/6486
Inventor 秦雷陈晓青朱丹丹张吉顺左楠楠彭雪婷
Owner 江苏雷森生物科技有限公司
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