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Norovirus GI and GII type quantum dot joint inspection test strip as well as preparation method and application thereof

A technology of quantum dots and test strips, which is applied in the field of virus GI and GII quantum dot joint detection test strips and its preparation, can solve the problems of joint detection of norovirus GI and GII antigens that have not yet been seen, and achieve protection stability , high sensitivity, the effect of improving accuracy and sensitivity

Active Publication Date: 2020-04-03
AFFILIATED HOSPITAL OF GUANGDONG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no related product based on the joint detection of norovirus GI and GII antigens based on quantum dot labeling

Method used

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  • Norovirus GI and GII type quantum dot joint inspection test strip as well as preparation method and application thereof
  • Norovirus GI and GII type quantum dot joint inspection test strip as well as preparation method and application thereof
  • Norovirus GI and GII type quantum dot joint inspection test strip as well as preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0042] Embodiment 1, a kind of norovirus GI and GII type antigen quantum dot joint detection test strip

[0043] The test strip structure of this embodiment is as follows: figure 1 As shown, it includes a bottom plate 1, a sample pad 2, a binding pad 3, a nitrocellulose membrane 4, and a water-absorbing pad 5 overlapped successively on the bottom plate from left to right, and the binding pad is coated with quantum dot-labeled Nutraceuticals. Such as virus GI type detection antibody and quantum dot-labeled Norovirus GII type detection antibody; the nitrocellulose membrane is provided with 401 detection line 1, 402 detection line 2 and quality control line 403, and the detection line 1 is coated with There is norovirus type GI capture monoclonal antibody, and the detection line 2 is coated with norovirus type GII capture monoclonal antibody; the quality control line is coated with rabbit anti-mouse IgG polyclonal antibody.

[0044] Wherein, in this embodiment, the sample pad an...

Embodiment 2

[0046] Embodiment two, the preparation method of test strip of the present invention

[0047] (1) Preparation of sample pad

[0048] Cut the sample pad into an appropriate size and length that matches the binding pad, spray the sample pad treatment solution on the sample pad at an amount of 40 μL / cm, soak it at room temperature for 2 hours, and dry it in a 37°C thermostat for no less than 16 hours; Drying for use; the sample treatment solution is a phosphate buffer (0.01M, pH 7.2 ).

[0049] (2) Preparation of binding pads

[0050] ①Take 1 mg of carboxyl water-soluble quantum dots, and then suspend the quantum dots in 800 μL of MES buffer solution with a concentration of 0.05M and a pH of 6.0;

[0051] ②Use 0.05M pH6.0 MES buffer to prepare NHS (50mg / ml) and EDC (50mg / ml), take 20μl each and add to quantum dots, the final concentration is NHS (1mg / ml) and EDC (1mg / ml) ;

[0052] ③After activation for about 20 minutes, centrifuge and wash, 16000g at 4°C, 10min×2 times, dis...

Embodiment 3

[0064] Embodiment 3, the screening experiment of quantum dot complex solution

[0065] Adopt different quantum dot complex solutions to resuspend quantum dot-labeled Norovirus GI type detection antibodies and quantum dot-labeled Norovirus GII type detection antibodies, process 1: quantum dot complex solutions contain 6 wt% of Turanose, Tween -40 0.08wt%, glycine 0.9wt% and ProClin950 0.06v% phosphate buffer (0.01M, pH 7.2); treatment 2: quantum dot complex solution containing Tween-40 0.08wt%, glycine 0.9wt% and ProClin950 0.06v% phosphate buffer (0.01M, pH 7.2); treatment: 3: Quantum dot recombination solution is phosphate buffer containing 6wt% turose, Tween-40 0.08wt% and ProClin950 0.06v% (0.01M, pH 7.2). The above-mentioned different quantum dot complex solutions were used to prepare each group of test strip samples according to the method described in Example 2, and each group of test strip samples prepared above was added dropwise to the negative control group for dete...

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Abstract

The invention discloses a Norovirus GI and GII type quantum dot joint inspection test strip as well as a preparation method and application of the Norovirus GI and GII type quantum dot joint inspection test strip. The test strip comprises a bottom plate, and a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad which are sequentially overlapped on the bottom plate from left to right, and the combination pad is coated with a quantum dot labeled Norovirus GI type detection antibody and a quantum dot labeled Norovirus GII type detection antibody. The test strip ofthe invention uses the quantum dots as markers, can be coupled with biomolecules easily, and is a novel rapid diagnosis and detection marker and has high fluorescence intensity and strong stability, so that the detection is higher in sensitivity, and the early screening of enterovirus infection and other diseases is convenient. The test strip is combined with an immunochromatography technology tobe prepared into a strip-shaped structure, is light and easy to carry, does not need the complex equipment, and is very suitable for the field screening and bedside detection.

Description

technical field [0001] The invention belongs to the technical field of medical testing, and in particular relates to a norovirus GI and GII type quantum dot joint detection test strip and a preparation method and application thereof. Background technique [0002] Norovirus (Norovirus, Novs) was first discovered and named as Norwalk virus (norwalkvirus, NV) by American scholar Kapikian in the feces of patients with diarrhea in Norwalk Town in 1972, and the virus was subsequently named Norovirus. Norovirus is a small non-enveloped virus with a diameter between 27-40nm and a single-stranded RNA genome of approximately 6.5-7.5kb, classified in the Norovirus genus of the family Caliciviridae. Based on the complete amino acid sequence analysis of the capsid protein, it can be subdivided into at least 33 genotypes (9 GI, 22 GII and 2 GIV genotypes). The genotypes of noroviruses that infect humans are mainly GI, GII, and GIV, which can cause acute gastroenteritis, and the main clin...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/569G01N33/531G01N33/558
CPCG01N33/531G01N33/558G01N33/56983G01N33/577G01N2333/08
Inventor 潘庆军吴平张莉芳蒋娜王思捷李新新吴晗
Owner AFFILIATED HOSPITAL OF GUANGDONG MEDICAL UNIV
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