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Kit suitable for rapidly detecting AMH and INHB by using double-tagging time resolution fluorescence immunoassay method and use method of kit

A time-resolved fluorescence and immunoassay technology, which is used in biological testing, fluorescence/phosphorescence, measurement devices, etc., and can solve the problems of large amount of blood samples, cumbersome detection steps, and long sample addition time.

Inactive Publication Date: 2015-06-24
GUANGZHOU FENGHUA BIOENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The purpose of the present invention is to solve the problems in the prior art that a kit can only detect AMH or INHB alone, the sensitivity is low, the detection steps are cumbersome, the sample addition time is long, and the amount of blood samples is large

Method used

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  • Kit suitable for rapidly detecting AMH and INHB by using double-tagging time resolution fluorescence immunoassay method and use method of kit
  • Kit suitable for rapidly detecting AMH and INHB by using double-tagging time resolution fluorescence immunoassay method and use method of kit
  • Kit suitable for rapidly detecting AMH and INHB by using double-tagging time resolution fluorescence immunoassay method and use method of kit

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0047] The preparation method of the kit in the above improvement scheme includes the following steps:

[0048] 1) Prepare a solid phase carrier coated with AMH Pcab and INHBβMcab; 2) Prepare a mixed calibrator of AMH and INHB; 3) Prepare Eu 3+ -INHαMcab; 4) Preparation of Sm 3+ -AMH Mcab; 5) Prepare immune response promoting solution, concentrated lotion and enhancement solution; 6) Dispense the mixed calibrator of AMH and INHB, Eu 3+ -INHαMcab, Sm 3+ -AMH Mcab, immune response promoting liquid, concentrated lotion and enhancement liquid; 7) labeling; 8) assembly into finished products.

[0049] Wherein, in step 1), the following steps are used to prepare the solid phase carrier coated with AMH Pcab and INHBβMcab:

[0050] Use AMH Pcab and INHBβMcab with coating buffer (50mmol / L, pH 9.6 carbonic acid buffer, 20mmol / L, pH 4.5 phosphate buffer, 50mmol / L, pH 7.8 Tris-HCl buffer or 50mmol / L, pH 4.5 citrate buffer, etc.) to the optimum concentration, coating on the solid-phase carrier, ...

Embodiment 1

[0082] Reagent kit suitable for detecting AMH and INHB using double-labeled rapid time-resolved fluorescence immunoassay and preparation thereof Method and usage

[0083] A kit suitable for the detection of AMH and INHB using double-labeled rapid time-resolved fluorescence immunoassay, including:

[0084] 1) Antibody solid phase carrier: a solid phase carrier coated with AMH Pcab and INHBβMcab;

[0085] 2) AMH and INHB mixed calibrators: calibrators B, C, D, E, F contain AMH standard raw materials and INHB standard raw materials;

[0086] 3) Lanthanide markers: marker 1 contains Eu 3+ -INHαMcab, marker 2 contains Sm 3+ -AMH Mcab;

[0087] 4) Immune response promoting solution: containing 100ml / L BSA, 0.05g / L eosin, 6ml / L Tween20, 0.01mg / L EDTA, 30mg / L Procline300, and pH7.8, 50mmol / L Tris containing 4% PEG 6000 -HCl buffer.

[0088] 5) Concentrated washing solution: Contains Tween20 and brilliant blue, Procline300, Tris-HCl buffer;

[0089] 6) Enhancement solution: contains Triton, gla...

Embodiment 2

[0125] The detection time of the double-labeled fast time-resolved fluorescence immunoassay and the traditional time-resolved fluorescence immunoassay of the present invention Compare

[0126] The detection time comparison between the double-labeled fast time-resolved fluorescence immunoassay of the present invention and the traditional time-resolved fluorescence immunoassay is shown in the following table.

[0127]

[0128]

[0129] It can be seen from the above table that the double-labeled fast time-resolved fluorescence immunoassay can simultaneously detect AMH and INHB in the same solid-phase carrier reaction site, and the double-labeled fast time-resolved fluorescence immunoassay only needs 1 / of the detection time of the traditional time-resolved fluorescence immunoassay. Within 4 hours, it saves nearly one and a half times compared to the single-label rapid method, which is shorter than the existing enzyme-linked immunoassay method (1~2h).

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Abstract

The invention provides a kit suitable for rapidly detecting AMH and INHB by using a double-tagging time resolution fluorescence immunoassay method. The kit comprises (1) a solid phase carrier coated by an AMH coating antibody and an INHB coating antibody; (2) a mixed calibration product of AMH and INHB; (3) an AMH detection antibody marked by using a lanthanide element 1; (4) an INHB detection antibody marked by a lanthanide element 2; (5) an immunoreaction accelerant liquid; (6) a concentrated washing liquid; and (7) a reinforcing liquid. The invention further provides a use method of the kit for rapidly detecting AMH and INHB by using the double-tagging time resolution fluorescence immunoassay method. By adopting a double-tagging time resolution fluorescence immunoassay technique, the defect that conventional AMH and INHB respectively need a single kit, that is, one kit can be only used for detecting AMH or INHB can be overcome, the detection steps can be reduced, the sampling time can be shortened, and the use amount of a blood sample can be reduced.

Description

Technical field [0001] The invention relates to a double-labeled rapid time-resolved fluorescence immunoassay kit, in particular to a reagent kit suitable for rapid detection of AMH and INHB by using the double-labeled time-resolved fluorescence immunoassay and a method of use thereof. Background technique [0002] According to the World Health Organization (WHO), fertility problems affect about 10% of couples, affecting as many as 80 million people worldwide, and this number is still rising. The ability of the normal female reproductive system to conceive is called fertility potential. Ovarian reserve (OR) reflects a woman's fertility. Decreased OR means that the number of remaining eggs in the ovary drops to a threshold, which affects the fertility potential and leads to low fertility. Because OR is closely related to the diagnosis and treatment of fertility and infertility, it has attracted increasing attention. For women who have not completed the fertility event and are ab...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N21/64
CPCG01N33/582G01N33/54393G01N33/74
Inventor 谭玉华
Owner GUANGZHOU FENGHUA BIOENG
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