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HIV p24 high-sensitivity detection method based on quantum dot selective cation exchange reaction

A technology of cation exchange and detection method, which is applied in the direction of immunoassay, measuring device, instrument, etc., can solve the problems of high cost, low analytical sensitivity of ultraviolet absorption spectrometer, difficulty in realizing portable visual rapid analysis, etc., achieve high long-term stability, improve Effects of analytical sensitivity and high luminous efficiency

Pending Publication Date: 2021-01-29
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above electrochemiluminescence strategies have high instrument costs, low analytical sensitivity of ultraviolet absorption spectrometers, and are easily interfered by the detection medium; the most important thing is that they all need to rely on existing detection instruments, and it is difficult to achieve portable visual rapid analysis

Method used

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  • HIV p24 high-sensitivity detection method based on quantum dot selective cation exchange reaction
  • HIV p24 high-sensitivity detection method based on quantum dot selective cation exchange reaction
  • HIV p24 high-sensitivity detection method based on quantum dot selective cation exchange reaction

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Experimental program
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Effect test

Embodiment 1

[0052](1) Add 10μL of 20μg / mL recognition antibody (Ab1) Was mixed with 30μL of analysis buffer and added to a 96-well plate, and incubated overnight at 4°C. Wash the 96-well plate 3 times with 200 μL of washing buffer to remove unbound recognition antibody. Subsequently, 100 μL of blocking buffer was added and incubated at 37°C for 1 hour to seal the remaining vacancies in the 96-well plate, and washed 3 times. Subsequently, 40 μL of the serum sample to be tested and 60 μL of analysis buffer were added to the 96-well plate, incubated at 37° C. for 1 hour, and washed three times. 40μL of 10μg / mL biotin-labeled detection antibody (Ab2) Was added to a 96-well plate together with 60 μL of assay buffer, and incubated at 37°C for 1 hour to form a primary antibody-p24-detection antibody-biotin sandwich complex, and washed three times.

[0053](2) Add 40 μL of 5 μg / mL streptavidin-alkaline phosphatase (SA-ALP) and 60 μL of analysis buffer to the 96-well plate obtained in step (1), incubate at...

Embodiment 2

[0057](1) Add 10μL of 30μg / mL recognition antibody (Ab1) Was mixed with 10μL of analysis buffer and added to a 96-well plate, and incubated overnight at 2°C. Wash the 96-well plate twice with 200 μL of washing buffer to remove unbound recognition antibody. Subsequently, 100 μL of blocking buffer was added and incubated at 37° C. for 1 hour to seal the remaining vacancies in the 96-well plate, and washed twice. Subsequently, 40 μL of the serum sample to be tested and 40 μL of analysis buffer were added to the 96-well plate, incubated at 35° C. for 80 min, and washed three times. 40μL of 5μg / mL biotin-labeled detection antibody (Ab2) Was added to a 96-well plate together with 40 μL of assay buffer, and incubated at 35°C for 80 min to form a primary antibody-p24-detection antibody-biotin sandwich complex, and washed twice.

[0058](2) Add 40 μL of 2 μg / mL streptavidin-alkaline phosphatase (SA-ALP) and 40 μL of assay buffer to the 96-well plate obtained in step (1), incubate at 20°C for 20...

Embodiment 3

[0061](1) Add 10μL of 10μg / mL recognition antibody (Ab1) Was mixed with 50μL of analysis buffer and added to a 96-well plate, and incubated overnight at 6°C. Wash the 96-well plate 4 times with 200 μL of washing buffer to remove unbound recognition antibody. Subsequently, 100 μL of blocking buffer was added and incubated at 37° C. for 1 hour to seal the remaining vacancies in the 96-well plate, and washed 4 times. Subsequently, 40 μL of the serum sample to be tested and 80 μL of analysis buffer were added to the 96-well plate, incubated at 39° C. for 40 min, and washed three times. 40μL of 15μg / mL biotin-labeled detection antibody (Ab2) Add 80μL of assay buffer to a 96-well plate and incubate at 39°C for 40min to form a primary antibody-p24-detection antibody-biotin sandwich complex, and wash 4 times.

[0062](2) Add 40 μL of 8 μg / mL streptavidin-alkaline phosphatase (SA-ALP) and 80 μL of analysis buffer to the 96-well plate obtained in step (1), incubate at 30°C for 10 minutes and was...

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Abstract

The invention relates to an HIV p24 high-sensitivity detection method based on a quantum dot selective cation exchange reaction. The HIV p24 high-sensitivity detection method comprises the following steps: preparing an anti-p24-detection antibody-biotin sandwich compound by adopting a recognition antibody, a to-be-detected sample and a detection antibody; connecting the anti-p24-detection antibody-biotin sandwich compound with streptavidin-alkaline phosphatase to obtain an anti-p24-detection antibody- ALP compound; and adding vitamin C-2-phosphate into the anti-p24-detection antibody-ALP compound for incubation, then adding a silver nitrate solution for a reaction, adding a quantum dot stock solution for the reaction, analyzing fluorescence signals, and acquiring a detection result. A light-emitting quantum dot selective cation exchange reaction signal amplification technology is introduced so that analysis sensitivity is improved; the color of the light-emitting quantum dots is in a visible light range so that visual analysis of human eyes is easy to realize; and the fluorescence analysis sensitivity is high, a sample demand is low, an anti-interference capability is high, an analysis speed is high, and the miniaturized fluorophotometer is convenient to carry.

Description

Technical field[0001]The invention belongs to the field of biomedical diagnosis, and specifically relates to a high-sensitivity HIV p24 detection method, kit and application thereof based on the selective cation exchange reaction of quantum dots.Background technique[0002]AIDS (AIDS) is a major infectious disease caused by HIV that seriously threatens human health. Its death rate and mortality rate rank first among legal infectious diseases in my country. According to the World Health Organization, as of 2018, there were more than 37.9 million people living with HIV (PLWH), and only 75% of them knew their infection status. HIV infection can be divided into three stages: acute HIV infection (AHI), asymptomatic infection and AIDS. Among them, AHI is the most dangerous stage of infection, and the risk of infection is 26 times that of the asymptomatic infection period. The 90-90-90 goals proposed by the UNAIDS / World Health Organization and the "Knowyour status" theme of World AIDS Day em...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/535G01N21/64
CPCG01N33/56988G01N33/535G01N21/6428G01N2333/161G01N2469/10G01N2021/6432Y02A50/30
Inventor 陈飘飘陶传敏应斌武唐卓芸耿佳
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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