Zika virus detection kit based on photonic crystal enhanced electrochemiluminescence and preparation method thereof

A detection kit and photonic crystal technology, applied in the field of biochemical analysis and detection, can solve problems such as low selectivity and poor sensitivity, and achieve the effects of improving sensitivity, improving detection evaluation, and reducing detection concentration and detection limit.

Active Publication Date: 2022-04-12
SOUTHEAST UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Purpose of the invention: In view of the poor sensitivity and low selectivity of Zika virus detection in the prior art, the present invention provides a Zika virus detection kit based on photonic crystal enhanced electrochemiluminescence, and provides a The preparation method of card virus detection kit

Method used

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  • Zika virus detection kit based on photonic crystal enhanced electrochemiluminescence and preparation method thereof
  • Zika virus detection kit based on photonic crystal enhanced electrochemiluminescence and preparation method thereof
  • Zika virus detection kit based on photonic crystal enhanced electrochemiluminescence and preparation method thereof

Examples

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Effect test

Embodiment 1

[0055] The ZIKV electrochemiluminescence immunosensor is prepared, and the immunosensor is used to detect ZIKV to verify the feasibility of the technical solution of the present invention.

[0056] (1) Preparation of photonic crystal thin film

[0057] ① SiO 2 Synthesis of microspheres

[0058] a. Mix 7mL TEOS with 42mL absolute ethanol, and ultrasonically obtain a homogeneous TEOS solution for later use;

[0059] b. Mix 6.3mL ammonia water, 42mL absolute ethanol and 14mL water, stir at 25°C, and rotate at 1800r / min;

[0060] c. Add the TEOS solution obtained in step a to the b solution, and after 1min, the rotation speed is reduced to 1000r / min, and stirred for 2h;

[0061] d. Wash by centrifugation, wash twice with water and ethanol respectively, and dry at 60°C to obtain SiO 2 Microspheres.

[0062] ② Synthesis of opal photonic crystal films by vertical deposition self-assembly method

[0063] First, clean the ITO with acetone, absolute ethanol, and deionized water, t...

Embodiment 2

[0082] This example evaluates the reproducibility of the ZIKV electrochemiluminescence immunosensor, and the results can only be reproduced to show that the obtained data have certain reliability.

[0083] A number of ZIKV electrochemiluminescent immunosensors were prepared by referring to the method of Example 1: first soak the ITO of the prepared surface-modified photonic crystal film in an acetonitrile solution containing 1% APTES for 20 minutes to obtain an aminated photonic crystal film; then 10 μL 10 μg / mL ZiKa MAb 0095 was incubated on the aminated photonic crystal at 37°C for 1 h, washed with PBS buffer (0.10mol / L, pH 7.4) to remove excess ZIKV primary antibody, and washed with 30 μL PBS solution containing 1.0% BSA (0.10mol / L, pH 7.4) for 30min to block non-specific binding sites; add dropwise 10μL of PBS solution containing 100fg / mL ZIKV nucleoprotein antigen (NS), incubate at 37°C for 1h, and wash with PBS buffer; Add 10 μL of prepared Ru(bpy) dropwise 3 2+ -COOH-...

Embodiment 3

[0086] This example evaluates the specificity of the ZIKV electrochemiluminescent immunosensor. Only when the immunosensor responds to a specific antigen but does not respond to other interfering substances, the constructed immunosensor has application value.

[0087] (1) Construction of ZIKV electrochemiluminescence immunosensor

[0088] First soak the ITO surface-modified photonic crystal film in an acetonitrile solution containing 1% APTES for 20 minutes to obtain an aminated photonic crystal film; then incubate 10 μL of 10 μg / mL ZIKV primary antibody on the aminated photonic crystal at 35°C for 2 hours , wash with PBS buffer (0.10mol / L, pH 7.4) to remove excess ZIKV primary antibody, and incubate with 30 μL of 1.0% BSA-containing PBS solution (0.10mol / L, pH 7.4) for 30 min to block non-specific binding sites ; Add 10 μL 100fg / mL ZIKV nucleoprotein antigen (NS) dropwise, incubate at 35°C for 2 hours, wash with PBS buffer; add 10 μL Ru(bpy) dropwise 3 2+ -COOH-labeled ZIKV...

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Abstract

The invention discloses a Zika virus detection kit based on photonic crystal enhanced electrochemiluminescence and a preparation method thereof. The detection kit includes Ru(bpy) based on photonic crystal and ZIKV as antigen 3 2+ ‑COOH-labeled sandwich ZIKV electrochemiluminescence immunosensor. The preparation method comprises the following steps: preparing a photonic crystal film with ZIKV primary antibody connected on the surface, and sealing its non-specific binding site; preparing Ru(bpy) 3 2+ ‑COOH-labeled ZIKV secondary antibody; the photonic crystal film shows the linked ZIKV primary antibody and Ru(bpy) 3 2+ ‑COOH-labeled ZIKV secondary antibodies were linked to ZIKV antigens, respectively. The Zika virus detection kit of the present invention adopts the ZIKV electrochemiluminescent immunosensor, which provides a new idea for the detection of Zika virus, and can detect the concentration of Zika virus easily and quickly, and the electrochemiluminescence is enhanced by photonic crystals. As a result, the detection signal of the electrochemiluminescence probe is enhanced, the detection sensitivity of the electrochemiluminescence immunosensor is improved, and the minimum detection concentration and detection limit are reduced.

Description

technical field [0001] The invention relates to a Zika virus detection kit based on photonic crystal enhanced electrochemiluminescence and a preparation method thereof, belonging to the technical field of biochemical analysis and detection. Background technique [0002] Zika virus disease (ZIKV Virus Disease) is a self-limiting acute disease caused by Zika virus (ZIKV Virus) and transmitted by mosquitoes, mainly through the bite of Aedes aegypti. The clinical features are mainly fever, rash, arthralgia or conjunctivitis, and rarely cause death. Aedes mosquitoes, which can transmit Zika virus, exist in some parts of southern my country. In recent years, imported dengue epidemics with a similar transmission mode have continued to increase, and have caused large-scale outbreaks in some southern provinces. With the increasingly close personnel exchanges with countries or regions related to the epidemic, there is a risk of Zika virus importation in my country. [0003] Currentl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/76G01N33/68
CPCG01N21/76G01N33/6854
Inventor 丁收年朱凯迪陆天赵春芹徐来弟王晓燕杜凤峦张慧君李真真
Owner SOUTHEAST UNIV
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