Fluorescence microballoon immunochromatography testing card for testing HIV and preparation method
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A technology of HIV and fluorescent microspheres, applied in the field of medical detection, to achieve the effect of shortening the window period, easy operation and low price
Inactive Publication Date: 2009-11-11
WUXI ZODOLABS BIOTECH
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Problems solved by technology
At present, many kinds of HIV detection reagents have been developed at home and abroad, but there are few reagents that can simultaneously take into account sensitivity, specificity, and easy and fast operation.
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Embodiment 1
[0050] 1. Preparation of fluorescein isothiocyanate-silica-shell-core dual-structure fluorescent microspheres
[0051] Add 4ml of absolute ethanol to 1mg of FITC and APS at a molar ratio of 1:10, and stir at 200rpm in the dark for 8 hours, then add 60ml of absolute ethanol, 2ml of ammonia water, 3ml of ethyl orthosilicate and 1ml of ultrapure water and mix for 8 hours. Then add Na 2 SiO 3 Closed microspheres, Na 2 SiO 3 The final concentration was 3%, blocked for 8 hours, centrifuged to wash the microspheres three times, and then ultrasonically dispersed the microspheres with 10ml of absolute ethanol. Add 60ml of absolute ethanol, 1ml of ammonia water, 1ml of ultrapure water, 50ul of MPS into a three-necked flask, add the microspheres after ultrasonic treatment, and stir at 25°C and 300rpm for 8 hours. Mix 0.07g SDBS, 0.05g NaHCO 3 , dissolved in 100ml of ultrapure water, added microspheres, passed nitrogen, 70°C, stirred at 300rpm, then added 500uL of styrene and 100uL o...
Embodiment 2
[0071] The preparation method of the present embodiment is basically the same as that of the first embodiment, the difference is that:
[0072] The fluorescent microspheres prepared in step 1 are dichlorotris(1,10-phenanthrene) ruthenium fluorescein-silicon dioxide shell-core dual-structure microspheres, and the preparation method is as follows:
[0073] Mix 3mg of dichlorotris(1,10-phenanthrene) ruthenium fluorescein, 60ml of absolute ethanol, 4ml of ammonia water, 2ml of ethyl orthosilicate and 3ml of ultrapure water for 8 hours, then add Na 2 SiO 3 Closed microspheres, Na 2 SiO 3 The final concentration was 3%, blocked for 8 hours, centrifuged to wash the microspheres three times, and then ultrasonically dispersed the microspheres with 10ml of absolute ethanol. Add 60ml absolute ethanol, 1ml ammonia water, 1ml ultrapure water, 50ul MPS and ultrasonically treated microspheres into a three-necked flask, stir at 25°C and 300rpm for 8 hours. Dissolve 0.07g SDBS in 100ml pH7...
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Abstract
The invention discloses a fluorescence microballoon immunochromatography testing card for quantitatively testing HIV and preparation method. The testing card comprises A and B test paper, a sample cushion, a glass fibrous membrane, a nitrocellulose membrane and drinking paper, wherein, the nitrocellulose membrane is thereon fixed with a test line and a quality control line for realizing the simultaneous test of HIV antibody and HIV p24 antigen. The invention takes nucleocapsid dual-structural light-emitting nano-particles compounded by silicon dioxide and fluorescent substance as marks and adopts immunochromatographic technique to realize quantitative immunoassay of HIV. In the testing process, fluorescence microballoon excitation light source is adopted to carry out excitation, after the emitted fluorescence passes through an optical filter device, all emission spectra are collected, aggregated and multiplied by CCD scanning technique or optical fiber technique, and are then converted into numerical signals, the concentration of the substance to be measured is automatically calculated by using the built-in analysis software in a fluorescence analyzer. The invention has the advantages of high sensitivity, precise quota, fast detection, convenient operation, and economy and practicality.
Description
technical field [0001] The invention belongs to the field of medical detection, and in particular relates to a detection card for quantitative detection of AIDS virus by using fluorescent microsphere immunochromatography technology and a preparation method thereof. Background technique [0002] AIDS is the English transliteration of Acquired Immunodeficiency Syndrome (AIDS). It is a malignant infectious disease caused by human immunodeficiency virus (human immunodeficiency virus, referred to as HIV, commonly known as AIDS virus). HIV is a retrovirus, and two types of HIV-1 and HIV-2 have been discovered. The main routes of transmission of AIDS are blood transmission, sexual transmission between heterosexuals and homosexuals, and vertical transmission between mother and child. [0003] Since the first case of AIDS was discovered in Los Angeles, USA in 1981, AIDS has spread rapidly all over the world. So far, more than 60 million people around the world have been infected w...
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