Long-optical-path enzyme-linked immunoassay for testing thyroid stimulating hormone, and kit

A thyroid-stimulating hormone and long optical path technology, which is applied in the direction of analytical materials, biological testing, measuring devices, etc., can solve problems such as insufficient sensitivity, achieve the effects of expanding the measurement range, increasing the amount of coated antibodies, and improving the performance of immunoassays

Active Publication Date: 2011-09-21
HTA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to overcome the shortcomings of insufficient sensitivity of the existing enzyme-linked immunoassay method, the present invention provides a long optical path optical fiber spectroscopy enzyme-linked immunoassay method, based on this method provides a long optical path optical fiber spectroscopy enzyme-linked immunoassay method for measuring TSH, And provide a TSH long optical path optical fiber spectroscopic enzyme-linked immunoassay kit for implementing the above method, which is used to meet the requirements for detecting human serum samples

Method used

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  • Long-optical-path enzyme-linked immunoassay for testing thyroid stimulating hormone, and kit
  • Long-optical-path enzyme-linked immunoassay for testing thyroid stimulating hormone, and kit
  • Long-optical-path enzyme-linked immunoassay for testing thyroid stimulating hormone, and kit

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preparation example Construction

[0097] The preparation method of above-mentioned kit comprises the following steps:

[0098] (1) Preparation of antibody-coated tubes

[0099] Dilute the TSH antibody to 0.6 mg / L with 0.03 M, pH 7.2 phosphate buffer, add 200 μL to each polystyrene tube, and keep overnight at 4 °C; discard the reaction solution, and add 1% bovine serum white to each tube Protein 0.03M pH 7.2 phosphate buffer 300ml, block at 37°C for 2 hours, discard the blocking solution, freeze or dry at room temperature, store at 4°C for use;

[0100] (2) Preparation of TSH series standard products

[0101] Dilute the pure TSH product with calf serum containing 0.2% thimerosal and inactivate at 56°C for 1.0 hour to a series of standard products with concentrations of 0, 0.05, 0.3, 4.0, 10, and 20 mIU / L, and pack in 1.0 ml per bottle , stored at -20°C for use;

[0102] (3) Preparation of enzyme markers

[0103] Weigh 5mg of horseradish peroxidase, dissolve in 1ml of distilled water, fully dissolve; add 1ml...

Embodiment 1

[0130] The TSH long-path ELISA method provided by the present invention is compared with the conventional TSH ELISA method: the conventional TSH ELISA method is established by using the same antibody as in the present invention, and the sensitivity is 0.06mIU / L after methodological optimization. , the linear range is 0.3~20mIU / L; the sensitivity of the analytical method of the present invention is 0.015mIU / L, and the linear range is 0.05~20mIU / L; the sensitivity of the analytical method of the present invention is improved by 4 times compared with the conventional ELISA method, and has obvious Sensitivity, linear range advantage (see Table 2 below). The linear range in the table refers to the range between the standard point where the first concentration is not zero and the standard point of the highest concentration; the signal-to-noise ratio refers to the ratio between the standard point where the first concentration is not zero and the zero standard point; Linear correlatio...

Embodiment 2

[0134] According to the TSH analysis method and operation steps of the present invention, a 0.5m fused silica tube was used as a flow sample cell to carry out an immunoassay experiment to obtain a better standard curve, sensitivity and signal-to-noise ratio (see Table 3).

[0135] Table 3 Experimental results of 0.5m fused silica tube flow sample cell

[0136]

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Abstract

The invention discloses long-optical-path enzyme-linked immunoassay for testing thyroid stimulating hormone and provides a kit of the method. The long-optical-path enzyme-linked immunoassay comprises the following test steps of: performing immune reaction, separating, developing, measuring and processing data. A long-optical-path flowing sample pool and an optical fiber spectrometer are adopted for measurement; the detection sensitivity is 0.015mIU / L by increasing a measuring optical path, adopting a low-background substrate solution as well as optimizing a component preparation method; and compared with the sensitivity of the conventional optimized enzyme-linked immunoassay in which the same antibody is used, the sensitivity is enhanced by four times. The test linear range is 0.05 to 20mIU / L. The immune reaction is performed in a wrapped tube, and the reaction mode is a dual antibody sandwich one-step method. The kit comprises an antibody coated tube, a serial calibrator, an enzyme marker, a condensed cleaning solution, the low-background substrate solution, a terminating solution and an end-product diluent. The method is high in sensitivity and proper in test range, and the requirement of clinical detection can be met; meanwhile, the kit has low manufacturing cost and is favorable to popularization and application.

Description

technical field [0001] The invention relates to the technical field of in vitro immunoassay, in particular to a long-light-path enzyme-linked immunoassay method and a kit for measuring Thyroid Stimulating Hormone (TSH). Background technique [0002] TSH is a basic glycoprotein hormone secreted by the anterior pituitary gland to promote the growth and function of the thyroid gland. Contains 211 amino acids. Sugars account for about 15% of the whole molecule, which is composed of two peptide chains - α chain and β chain, with a molecular weight of 25000-28000D. The amino acid sequence of the α-subunit of the TSH molecule is substantially identical to that of other glycoprotein hormones such as luteinizing hormone (LH), follicle-stimulating hormone (FSH) and human chorionic gonadotropin (HCG). The β subunit is unique to TSH and determines the specificity of TSH. In addition to the biologically active whole molecule TSH, there are biologically inactive α and β subunits in ser...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/31G01N33/78G01N33/543
Inventor 燕强奋官国英潘玉婷
Owner HTA CO LTD
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