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Human prolactin (PRL) detection kit, as well as preparation method and application thereof

A technology for detecting kits and prolactin, which is applied in biological testing, material inspection products, etc., can solve the problems of poor repeatability of enzyme immunoassay, short shelf life, environmental pollution, etc., and achieve good amplification effect, high accuracy, and good sensitivity Effect

Inactive Publication Date: 2018-06-15
CHEMCLIN DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The problem to be solved by the present invention is to provide a serum prolactin (Prolactin) homogeneous luminescent immunoassay kit and its preparation and use method, to solve the cumbersome washing steps of the existing heterogeneous immunoassay kit; Environmental pollution, short shelf life defects, overcome the poor reproducibility of enzyme immunoassay, prone to "hook effect" defects

Method used

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  • Human prolactin (PRL) detection kit, as well as preparation method and application thereof
  • Human prolactin (PRL) detection kit, as well as preparation method and application thereof
  • Human prolactin (PRL) detection kit, as well as preparation method and application thereof

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preparation example Construction

[0085] The first aspect of the present invention provides a method for preparing a human prolactin homogeneous detection kit, the method comprising:

[0086] preparing a first component, the first component comprising a receptor capable of reacting with singlet oxygen to generate a detectable signal, and a first binding unit bound thereto, the first binding unit capable of binding human lactation The first epitope of the element;

[0087] preparing a second component, the second component comprising the first label and a second binding unit bound thereto, the second binding unit is capable of binding a second epitope of human prolactin, the second epitope The first epitope is an epitope with different binding characteristics of human prolactin or an epitope with the same binding characteristics at a different position;

[0088] A third component is prepared comprising a donor capable of generating singlet oxygen in an excited state and bound thereto a specific binder of the f...

Embodiment 1

[0163] Embodiment 1-basic kit

[0164] The core components of the kit are:

[0165] The PRL quantitative assay detection kit (light-induced chemiluminescence method) of the present embodiment consists of a composition (reagent 1, R1) containing luminescent microspheres (FG-Ab1) coated with the first anti-PRL monoclonal antibody, containing biotin The composition (reagent 2, R2) of the labeled second anti-PRL monoclonal antibody (Bio-Ab2); also includes the sample to be tested. In addition, it includes a composition (reagent 3, R3) comprising photosensitive microspheres (SA-GG) coated with streptavidin, which is prepared according to the method of the aforementioned materials and equipment, including light-induced chemiluminescence analysis instrument and so on.

[0166] The using method of the test kit of the present embodiment comprises:

[0167] The detection process is fully automatically completed by the automatic light-induced chemiluminescence analysis system and the de...

Embodiment 2

[0176] Embodiment 2-establishment of standard function and quantitative kit

[0177] Preparation of kit materials:

[0178] The preparation methods of specific materials and corresponding reagents are prepared according to the materials and equipment section.

[0179] The using method of the test kit of the present embodiment comprises:

[0180] Reagents are added in parallel to different reaction wells, and the reaction wells are irradiated with excitation light with a wavelength of 680nm, and emission light with a wavelength of 612nm is detected. See Table 1 for specific concentrations and signal values ​​of the substances to be detected.

[0181] The specific detection steps are:

[0182] a. Add 25 μl samples to different reaction wells;

[0183] b. Add 25 μl R1 and 25 μl R2 in sequence to each reaction well;

[0184] c. Incubate at 37°C for 15 minutes;

[0185] d. Add 175 μl of R3 to each reaction well;

[0186] e. Incubate at 37°C for 15 minutes;

[0187] f. The l...

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Abstract

The invention discloses a preparation method of a human prolactin (PRL) homogeneous-phase detection kit. The method comprises the steps of preparing a component containing an acceptor and a first combination unit combined with the acceptor, wherein the acceptor can be reacted with singlet oxygen to generate a detectable signal, and the first combination unit can combine a first epitope of the PRL;preparing a second component containing a first marker and a second combination unit combined with the first marker, wherein the second combination unit can combine a second epitope of the PRL, and the second epitope and the first epitope are the epitopes with different combining characteristics of the PRL or the epitopes with same combining characteristics at different positions of the PRL; preparing a third component containing a donor and a specific conjugate combined with the first marker, wherein the donor can produce singlet oxygen at an excited state. According to the kit prepared through the method, the PRL can be high-efficiently detected.

Description

technical field [0001] The invention belongs to the technical field of biomolecular detection and relates to a homogeneous immunoassay kit without matrix effect, in particular to a homogeneous immunoassay kit for detecting human prolactin and its preparation method and application. Background technique [0002] Prolactin, also known as prolactin (PRL), is secreted by the anterior pituitary gland cells and is an essential condition for maintaining normal breast development and women's lactation. PRL has a molecular weight of about 22,500 Da and is a single-chain polypeptide composed of 198 amino acids containing 3 interchain disulfide bonds. The secretion of prolactin is mainly controlled by prolactin-inhibiting factor (dopamine) and prolactin-releasing factor (serotonin) released from the hypothalamus. Hypothalamic thyrotropin-releasing hormone (TRH) also stimulates the secretion of PRL, which can be used to detect the PRL stock and the challenge test of the abnormal secret...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/74
Inventor 赵宏强中华李会强刘宇卉李临
Owner CHEMCLIN DIAGNOSTICS CO LTD
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