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Laterally-flowing immunoassay method using time resolution up-converting phosphor technology

A technology of lateral flow chromatography and measuring device, which is applied in the detection field and can solve problems such as reducing the accuracy of results

Active Publication Date: 2013-09-18
何爱民
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a method for detecting the presence and content of an analyte in a sample, which solves the problem that the excitation light in the prior art is absorbed by the analyte itself and the liquid sample matrix in the near ultraviolet region, resulting in a significant decrease the accuracy of the results, etc.

Method used

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  • Laterally-flowing immunoassay method using time resolution up-converting phosphor technology
  • Laterally-flowing immunoassay method using time resolution up-converting phosphor technology
  • Laterally-flowing immunoassay method using time resolution up-converting phosphor technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Example 1: Preparation of up-conversion luminescent probes

[0083] Add a certain amount of ethanol to an appropriate amount (200 μl) of an aqueous solution of carboxylic acid functionalized polymethacrylic acid latex particles PP02N (0.33 μm in diameter) under stirring, to reach 65% of the total amount of solvent. An appropriate amount (eg, 1% by weight of the latex particles) of vinyl chloride containing the proprietary europium chelate (eg, 1% by weight of the total solvent) is slowly added to the particle suspension with stirring. The mixture was stirred for half an hour. An appropriate amount of water (eg three times the total volume of the initial solvent) is then slowly added to the stirred mixture over a certain period of time (eg 2 hours). After the addition of water was complete, most of the ethanol was removed from the mixture by rotary evaporator. The particles were then washed twice by centrifugation using 90% ethanol. The particles were then washed twic...

Embodiment 2

[0084] Example 2: Ligation of antibodies to upconverting luminescent probes to form conjugate probes

[0085] The carboxylic acid surface groups of the probe prepared in Example 1 were first activated by standard methods using carbodiimide. The transbiotin antibody was mixed with the activated probe for four hours. Probes were washed four times with Hepes buffer and suspended in Hepes buffer containing 10 mg / ml BSA and 0.5% Tween20 for storage.

Embodiment 3

[0086] Example 3: Time-resolved upconversion excitation and emission spectra of the conjugates prepared in Example 2

[0087] Suspend an appropriate amount of the conjugate prepared in Example 2 in water to prepare a particle suspension in cells (for example, a concentration of 10 ng / cell). Time-resolved upconversion excitation and fluorescence spectra were obtained using the following measurement parameters and are displayed in Figure 5 middle. For time-resolved upconversion fluorescence spectroscopy: excitation at 870 nm, sampling window 50 μs, each flash time 100 μs, initial delay 0.01 μs, number of flashes 10 times, number of scans 10 times, fluorescence collection from 500 nm to 800 nm. For time-resolved upconverted excitation spectra: emission at 615 nm, sampling window 50 μs, each flash time 100 μs, initial delay 0.01 μs, number of flashes 10, number of scans 10, excitation collected as Figure 5 Shown from 700nm to 900nm.

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Abstract

The invention discloses an immunoassay device, which can be used for detecting and measuring analyte, and can be used for detecting the time resolution phosphor signal by using the up-converting phosphor pin. The laterally-flowing immunoassay method using time resolution up-converting phosphor technology is more attractive than the traditional fluorescent laterally-flowing detecting method. A number reading device having advantages of simple structure and low costs can be designed, and can be used for the up-converting phosphor having the long service lifetime, and no expansive optical fiber and reflector are required. The number reading device can be used to generate the pulse IR photon by using the IR LED laser, and the pin captured by the laterally-flowing chromatography device can be triggered from one side; and the phosphor signal having the long service lifetime can be detected in the visible light area from the other side after the delay of the certain time period by using the silicon photoelectric diode.

Description

technical field [0001] The invention belongs to the technical field of detection, and specifically relates to a method and device for lateral flow immunochromatography determination using time-resolved up-conversion luminescence detection technology, and more specifically relates to a method for detecting whether an analyte exists in a sample and method for the determination of its content. Background technique [0002] Membrane-based lateral flow immunoassays (LFA) have been commercialized for the measurement of a large number of analytes. Examples of medical diagnostic products based on lateral flow immunoassay technology platforms include pregnancy tests and drug of abuse tests. Currently, these tests are mainly used in point-of-care (POC) and over-the-counter (OTC) testing markets due to their low cost and ease of use. Most existing commodities using the platform provide qualitative and semi-quantitative results. Recently, several lateral flow chromatographic assay te...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/63G01N33/558
CPCG01N33/558G01N33/54388
Inventor 何爱民
Owner 何爱民
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