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1. WO2022001090 - HOOK EFFECT DETERMINATION METHOD FOR HOMOGENEOUS TIME-RESOLVED FLUORESCENCE IMMUNOASSAY

Publication Number WO/2022/001090
Publication Date 06.01.2022
International Application No. PCT/CN2021/073019
International Filing Date 21.01.2021
IPC
G01N 21/64 2006.1
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using infra-red, visible or ultra-violet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
G01N 33/53 2006.1
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48Biological material, e.g. blood, urine; Haemocytometers
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
G01N 1/38 2006.1
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
1Sampling; Preparing specimens for investigation
28Preparing specimens for investigation
38Diluting, dispersing or mixing samples
CPC
G01N 1/38
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
1Sampling; Preparing specimens for investigation
28Preparing specimens for investigation ; including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
38Diluting, dispersing or mixing samples
G01N 21/6402
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
6402Atomic fluorescence; Laser induced fluorescence
G01N 21/6408
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
21Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
63optically excited
64Fluorescence; Phosphorescence
6408with measurement of decay time, time resolved fluorescence
G01N 33/53
GPHYSICS
01MEASURING; TESTING
NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
48Biological material, e.g. blood, urine
50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53Immunoassay; Biospecific binding assay; Materials therefor
Applicants
  • 中国科学院苏州生物医学工程技术研究所 SUZHOU INSTITUTE OF BIOMEDICAL ENGINEERING AND TECHNOLOGY, CHINESE ACADEMY OF SCIENCES [CN]/[CN]
Inventors
  • 刘涛 LIU, Tao
Agents
  • 北京三聚阳光知识产权代理有限公司 SUNSHINE INTELLECTUAL PROPERTY INTERNATIONAL CO., LTD.
Priority Data
202010603405.X29.06.2020CN
Publication Language Chinese (zh)
Filing Language Chinese (ZH)
Designated States
Title
(EN) HOOK EFFECT DETERMINATION METHOD FOR HOMOGENEOUS TIME-RESOLVED FLUORESCENCE IMMUNOASSAY
(FR) PROCÉDÉ DE DÉTERMINATION D'EFFET CROCHET DE DOSAGE IMMUNOLOGIQUE PAR FLUORESCENCE À RÉSOLUTION TEMPORELLE HOMOGÈNE
(ZH) 一种判断均相时间分辨荧光免疫分析钩状效应的方法
Abstract
(EN) The present application relates to a hook effect determination method for a homogeneous time-resolved fluorescence immunoassay. The method comprises the steps of: (1) measuring, under the same excitation wavelength, fluorescence intensities of a fluorescence donor and acceptor at two detection wavelengths, and using a fluorescence intensity ratio between the two wavelengths as a signal value; (2) respectively establishing, by means of measuring signal values of at least two time instances in a reaction initiation phase and performing linear fitting and four-parameter fitting, reference curves of an absolute signal intensity value, a signal change rate value, and a sample concentration; and (3) detecting a signal value of an unknown sample, and obtaining, by means of the reference curves, a concentration of the unknown sample according to a fluorescence intensity of the unknown sample, so as to avoid the hook effect. The invention enables calculation of a relative signal intensity and a relative signal change rate value by means of ratios of the absolute signal intensity value and signal change rate value to background signals, thereby achieving consistency of results across different instruments. The invention increases the number of sample concentration readings which can be completed within a given period of time, thereby improving the detection throughput of systems.
(FR) La présente demande se rapporte à un procédé de détermination d'effet crochet d'un dosage immunologique par fluorescence à résolution temporelle homogène. Le procédé consiste : (1) à mesurer, sous la même longueur d'onde d'excitation, des intensités de fluorescence d'un donneur et d'un accepteur de fluorescence à deux longueurs d'onde de détection, et à utiliser un rapport d'intensité de fluorescence entre les deux longueurs d'onde en tant que valeur de signal; (2) établir respectivement, au moyen de valeurs de signal de mesure d'au moins deux instances temporelles dans une phase de lancement de réaction et de la réalisation d'un ajustement linéaire et d'un ajustement à quatre paramètres, des courbes de référence d'une valeur d'intensité de signal absolue, d'une valeur de taux de changement de signal et d'une concentration d'échantillon; et (3) à détecter une valeur de signal d'un échantillon inconnu, et à obtenir, au moyen des courbes de référence, une concentration de l'échantillon inconnu en fonction d'une intensité de fluorescence de l'échantillon inconnu, de manière à éviter l'effet crochet. L'invention permet de calculer une intensité de signal relative et une valeur de taux de changement de signal relative au moyen de rapports de la valeur d'intensité de signal absolue et de la valeur de taux de changement de signal aux signaux d'arrière-plan, ce qui permet d'obtenir une cohérence de résultats sur différents instruments. L'invention augmente le nombre de relevés de concentration d'échantillon pouvant être réalisées dans une période donnée, ce qui permet d'améliorer le rendement de détection de systèmes.
(ZH) 本申请涉及一种判断均相时间分辨荧光免疫分析钩状效应的方法,步骤是:(1)测定荧光供体和受体在同一激发波长下、两个检测波长处的荧光强度,然后以两个波长的荧光强度比值作为信号值;(2)通过测量反应起始阶段至少两个时刻的信号值,通过线性拟合、四参数拟合,分别建立信号强度的绝对值、信号变化的速率值与样品浓度的标准曲线;(3)检测未知样本的信号值,通过标准曲线,根据未知样本的荧光强度,获得未知样本的浓度,从而避免hook效应。利用信号强度绝对值和信号变化速率值与背景信号的比值,计算相对信号强度和信号变化速率相对值,在不同仪器上实现一致的结果。可以在既定时间内完成更多样品的浓度判读,从而使系统具有更高检测通量。
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