Plate-type fluorometric enzyme immunoassay for peroxidase antibody and use thereof

A peroxidase and luciferase technology, applied in the field of medical ultra-trace detection, can solve the problems of staying, low specificity and sensitivity, and achieve the effects of convenient operation, high sensitivity and specificity, and easy clinical application.

Inactive Publication Date: 2009-05-13
GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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Problems solved by technology

[0004] The determination of hTPOAb in my country began in the 1980s, but it mostly stayed at the level of qualitative determination. The radioimmunoassay of thyroid microsomal antibody (TMAb) is still a relatively common detection method in China. However, since this method measures TMAb, It is not pure hTPOAb, so the specificity and sensitivity of TMAb detection are lower than hTPOAb detection

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  • Plate-type fluorometric enzyme immunoassay for peroxidase antibody and use thereof
  • Plate-type fluorometric enzyme immunoassay for peroxidase antibody and use thereof
  • Plate-type fluorometric enzyme immunoassay for peroxidase antibody and use thereof

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[0019] The peroxidase antibody plate-type fluorescent enzyme immunoassay method of the present invention uses human thyroid peroxidase (hTPO) protein as an antigen-coated porous plate, and seals it with PBST containing 1-5% BSA; hTPO primary antibody, enzyme-labeled secondary antibody, fluorescent substrate and stop solution, and finally measure the fluorescence intensity on a fluorescence / chemiluminescence analyzer.

[0020] The antigen coating of the multi-well plate is as follows: add 100ng-2μg / 100μl hTPO to each well of the multi-well plate, overnight at 4°C, then rinse, then block with PBST blocking solution containing 1-5% BSA, add 100μl to each well, 25 Incubate at -37°C for 1 hour, then rinse and complete.

[0021] The porous plate is a porous plastic plate with different numbers of holes.

[0022] The addition of the first anti-hTPO antibody is: add 100 μl to each well of a multi-well plate with a human serum sample diluted 1:50-1:200 with a diluent or a standard sub...

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Abstract

The invention relates to a peroxydase antibody plate type fluorescence enzyme lmmunoassay and the application thereof. The method is characterized in that human thyroid peroxydase (hTPO) protein is taken as antigens to cover a perforated plate, and PBST containing 1-5 percent BSA is used for sealing the perforated plate; the perforated plate is then taken as a carrier and is added with an anti-hTPO first antibody or a standard, an enzyme-labeled second antibody, fluorogenic substrate and stop solution in sequence; and finally, the fluorogenic strength is detected on a fluorogenic/chemical luminescent analyzer. The method can be used to clinically distinguish autoimmunity thyropathy from non-autoimmunity thyropathy and diagnosis lymphocytic thyroiditis. The invention has the advantages that the perforated plastic plate is taken as a solidoid carrier to carry out the fluorescence enzyme lmmunoassay, the plate material has easy sources, low cost and easy popularization; the method is special, sensitive and stable; the detection value is calibrated according to national standard with favorable accuracy and the method is applicable to clinically requirement and popularization.

Description

【Technical field】 [0001] The invention belongs to the technical field of medical ultra-trace detection, and uses a porous plate as a carrier to prepare a solid-phase antigen microplate plate for the establishment and clinical application of a fluorescent enzyme immunoassay (FEIA) for quantitative detection of human thyroid peroxidase (hTPO) antibody . 【Background technique】 [0002] Autoimmune thyroid disease (AITD) is a group of frequently-occurring diseases, mainly including Hashimoto's thyroiditis (HT), Graves' disease (GD), postpartum thyroiditis (PPTD) and so on. Its pathogenesis is affected by various factors such as genetic background, environmental conditions, and immune system, and is manifested as abnormal thyroid function, accompanied by the production of various thyroid autoantibodies, such as thyroid peroxidase antibody (hTPOAb), thyrotropin-affected antibody (TRAb), thyroglobulin antibody (TgAb), etc. [0003] hTPO and hTPOAb play an important role in the pat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N21/64G01N33/544G01N33/564
Inventor 方佩华张艳丽许静李宁吕枚
Owner GENERAL HOSPITAL OF TIANJIN MEDICAL UNIV
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