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Test paper strip and method for detecting canine coronavirus through immunofluorescence chromatography technique

A fluorescent immunochromatography, coronavirus technology, applied in the field of fluorescent immunochromatography to detect canine coronavirus, fluorescent immunochromatography technology to detect canine coronavirus test strips, can solve false negative and false positive, colloidal gold immunity The problems of low chromatographic sensitivity and insufficient stability can avoid serious injury.

Inactive Publication Date: 2016-07-06
杨挥
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional method for detecting canine coronavirus is colloidal gold immunochromatography, but the sensitivity of colloidal gold immunochromatography is low, the stability is not good enough, and false negatives and false positives are prone to occur

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  • Test paper strip and method for detecting canine coronavirus through immunofluorescence chromatography technique
  • Test paper strip and method for detecting canine coronavirus through immunofluorescence chromatography technique

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1, the preparation of the test strip that detects canine coronavirus antigen with fluorescent immunochromatography

[0031] figure 1 The preparation method of the test strip for detection of canine coronavirus by fluorescent immunochromatographic technique is as follows:

[0032] 1. Carboxyl-modified SiO containing europium complexes 2 The microspheres were provided by Changchun Zhiang Biotechnology Co., Ltd., and the canine coronavirus antibody and antigen were purchased from antibodies-online.

[0033] 2. Coupling of fluorescent microspheres and canine coronavirus antibody

[0034]Mix 200μl 20mMPBS, pH7.4, fluorescent microspheres with a solid content of 1% and 6 mg carbodiimide, shake at room temperature for 30 minutes, centrifuge at 12,000rpm for 10 minutes, wash twice with 0.1M borate buffer, and activate the Fluorescent microspheres were resuspended in 300 μl borate buffer. Add 200 μl of 1 mg / ml canine coronavirus antibody to the microsphere suspens...

Embodiment 2

[0039] Embodiment 2, detect canine coronavirus antigen with fluorescent immunochromatographic test strip

[0040] 100 μl of solutions containing 0ng / ml, 10ng / ml, 100ng / ml, 1000ng / ml, and 10000ng / ml canine coronavirus antigen were added dropwise to the fluorescent immunochromatography test strip sample pad prepared in Example 1, and the reaction was 15 Minutes later, use a fluorescence detector that emits 395nm excitation light (Shanghai Haiyue Plastic Mold Co., Ltd.) to detect the fluorescence signal, and at the same time, 100 μl containing 0ng / ml, 10ng / ml, 100ng / ml, 1000ng / ml, 10000ng / ml canine coronavirus The antigen solution was added dropwise to the colloidal gold immunochromatographic test strip, and the result was observed after reacting for 5 minutes. The results of the two methods were compared as figure 2

[0041] Depend on figure 2 It can be seen that when the concentration of canine coronavirus antigen is 0ng / ml, there is no signal on the T line (the position o...

Embodiment 3

[0042] Embodiment 3, detect canine coronavirus antigen with fluorescence immunochromatography kit

[0043] The fluorescent immunochromatography kit includes reagent bottles and test strips. The preparation method of the test strip is the same as in Example 1, except that the canine coronavirus detection antibody marked with fluorescent microspheres is not sprayed on the marker pad of the test strip. However, the vial of the kit contains a fluorescently labeled canine coronavirus antibody.

[0044] When testing, first add the sample solution containing canine coronavirus into the reagent bottle, mix it and add it dropwise on the above-mentioned test strip, and detect the fluorescent signal after reacting for 5 minutes. Compared with the results of colloidal gold immunochromatography figure 2 , with little difference from Example 2.

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Abstract

The invention relates to a test paper strip and method for detecting a canine coronavirus through immunofluorescence chromatography technique, and specifically discloses the test paper strip for detecting the canine coronavirus through immunofluorescence chromatography technique. The test paper strip includes a marker pad containing a fluorescent-microsphere-labeled canine coronavirus antibody, and a capture membrane containing another canine coronavirus antibody. The invention further discloses the method for detecting canine coronavirus through immunofluorescence chromatography technique. The method comprises: dropwise adding a canine-coronavirus-containing solution to the sample pad of the test paper strip, and detecting a signal through a strip reader after reaction for a while.

Description

technical field [0001] The invention relates to the fields of nanometer material technology, immunology technology, lateral flow chromatography technology and the like. More specifically, the present invention relates to a test strip for detecting canine coronavirus by fluorescent immunochromatography; in addition, the present invention also relates to a method for detecting canine coronavirus by fluorescent immunochromatography. Background technique [0002] Canine coronavirus can infect dogs of all breeds and ages, especially puppies. Infected dogs will develop symptoms of gastroenteritis, frequent vomiting, diarrhea, anorexia, depression, accompanied by leukopenia, rapid dehydration, and even death. Sometimes it will recur 14-21 days after the symptoms disappear, which seriously endangers the health of dogs. [0003] If a dog is infected with canine coronavirus, it needs to be detected and treated early. The traditional method for detecting canine coronavirus is colloi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
Inventor 杨挥
Owner 杨挥
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