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Feline parvovirus antibody fluorescence detection test strip and preparation method and application thereof

A feline distemper virus, fluorescence detection technology, applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve the problems of high test conditions and operation levels, restricting the epidemic of feline distemper virus, and high operation requirements, and achieves improved combination. Efficiency, high practical value and promotion value, the effect of improving sensitivity

Active Publication Date: 2019-05-17
BEIJING KWINBON BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The hemagglutination inhibition test requires fresh and highly sensitive red blood cells, and the detection time takes 2 to 3 hours; the neutralization test is cumbersome to operate, requires high test conditions and operating levels, and the whole test takes 1 to 2 weeks to produce results; ELISA The method has high requirements for operation and needs the results of the microplate reader, and is not suitable for the detection of a single sample; although the immunocolloidal gold method is convenient for detection and does not require professional technicians and working environment, its detection sensitivity is low, and it can only be qualitative. Narrow detection range
These all limit their application in the clinical detection of feline distemper virus antibody. Therefore, the purpose of the patent of the present invention is to overcome the shortcomings of the above methods, and to establish a kind of feline distemper virus antibody that is easy to operate, sensitive, specific and fast. Quantitative detection method, applied to the monitoring of feline distemper virus immune effect, thereby limiting the prevalence of feline distemper virus

Method used

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  • Feline parvovirus antibody fluorescence detection test strip and preparation method and application thereof
  • Feline parvovirus antibody fluorescence detection test strip and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: the composition of feline distemper virus antibody fluorescent detection test strip ( figure 1 )

[0028] The test strip is composed of a bottom plate (8), a sample absorbent pad (1), a conjugate release pad a (2), a conjugate release pad b (3), a nitrocellulose membrane (4), an absorbent pad (5) composition;

[0029] The sample absorbent pad (1), conjugate release pad a (2), conjugate release pad b (3), nitrocellulose membrane (4), and water absorbent pad (5) are pasted on the PS bottom plate (8) in sequence Above; 1 / 4 area of ​​the conjugate release pad a (2) from the starting end is covered by the sample absorbent pad (1), and 1 / 4 area of ​​the conjugate release pad b (3) from the starting end is covered by the conjugate release pad a ( 2) Covering, the end of the conjugate release pad b (3) is connected to the beginning of the nitrocellulose membrane (4), the end of the nitrocellulose membrane (4) is connected to the beginning of the absorbent pad (5...

Embodiment 2

[0031] Embodiment 2: the preparation of test strip described in embodiment 1

[0032] The preparation method of this test strip mainly comprises the following steps:

[0033] 1) Prepare a nitrocellulose membrane with a detection line coated with feline distemper virus VP2 expression protein and a quality control line coated with goat anti-chicken anti-antibody;

[0034] 2) Prepare a conjugate release pad a sprayed with chicken anti-cat anti-antibody-biotin conjugate and a conjugate release pad b sprayed with streptavidin-fluorescent microsphere marker;

[0035] 3) Fix the sample absorbent pad, conjugate release pad a, conjugate release pad b, nitrocellulose membrane, and water absorbent pad on the base plate in sequence.

[0036] The following step-by-step detailed description:

[0037] (1) Preparation of each component

[0038] 1. Expression and purification of feline distemper virus VP2 protein

[0039] (1) VP2 protein partial gene sequence selection

[0040]①DNA序列:atga...

Embodiment 3

[0074] Embodiment 3: the use of test strip described in embodiment 1

[0075] 1. Sample Preparation

[0076] (1) Plasma samples: collect blood with an anticoagulant tube, or add EDTA anticoagulant to the blood collection tube first, add the collected blood and shake it slowly, and centrifuge at 3000-5000r / min for 10-30min at room temperature (adjust according to the amount of blood collected). The upper layer of yellow clear liquid is drawn as the plasma sample.

[0077] (2) Serum sample: Add the collected blood into a blood collection tube (without anticoagulant), place it at an incline at room temperature, and centrifuge at 3000-5000 r / min for 10-30 minutes at room temperature when a yellow clear liquid is precipitated on the upper layer (adjust according to the amount of blood collected). The upper layer of clear yellow liquid is drawn as the serum sample.

[0078] (3) Sample dilution: Take the above-mentioned sample that has been returned to room temperature, accurately ...

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PUM

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Abstract

The invention discloses a feline parvovirus antibody fluorescence detection test strip and preparation method and application thereof. The test strip provided by the invention comprises a sample absorbent pad, a conjugate release pad a, a conjugate release pad b, a nitrocellulose membrane, a water absorbent pad and a bottom plate; the chick anti-cat anti-antibody-biotin conjugate is sprayed-coatedon the conjugate release pad a, a streptavidin-fluorescent microsphere maker is sprayed-coated on the conjugate release pad b, a detection line and a quality control line are arranged on the nitrocellulose membrane, the feline parvovirus VP2 expression protein is coated on the detection line and a goat anti-chick anti-antibody is coated on the quality control line. The test strip disclosed by theinvention has the advantages of being high in sensitivity, strong in specificity, simple in operation, economic and practical, and the quick and quantitative detection of the feline parvovirus antibody field can be realized.

Description

technical field [0001] The invention relates to the detection of feline distemper virus antibody, in particular to a feline distemper virus antibody fluorescence detection test strip and its preparation method and application. technical background [0002] Feline distemper virus disease is an acute and highly contagious fatal infectious disease caused by feline distemper virus (Feline parvovirus, FPV). In the natural state, FPV can infect almost all cats, mustelids and raccoons, causing high mortality in cats, minks, raccoons and other animals, and poses a serious threat to wild animals and special economic animals, so the diagnosis It is of great significance to prevent and control the prevalence of FPV in our country. FPV has a 20-plane stereosymmetric structure with a diameter of 20-24nm. The viral capsid protein is composed of three structural proteins: VP1, VP2 and VP3. Among them, FPV VP2 protein is the most conserved antigen-specific protein in FPV, and has strong i...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569G01N33/531
Inventor 万宇平崔廷婷肖文雪鲁晓雄张坤何方洋龙光宗李斌王兆芹曹东山
Owner BEIJING KWINBON BIOTECH