Kit for detecting serum amyloid A content and preparation method and detection method thereof

A technology for amyloid and serum amyloid, applied in the field of medical testing, can solve the problem of large sample demand and achieve the effect of less sample consumption, low detection limit and wide linear range

Inactive Publication Date: 2018-11-16
DIRUI MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods all use serum or plasma as the test sample, and all blood needs to be pre-separated from r

Method used

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  • Kit for detecting serum amyloid A content and preparation method and detection method thereof
  • Kit for detecting serum amyloid A content and preparation method and detection method thereof
  • Kit for detecting serum amyloid A content and preparation method and detection method thereof

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preparation example Construction

[0048] The present invention also provides a preparation method of a kit for measuring serum amyloid A content, comprising:

[0049] Step 1: Prepare the hemolytic agent R1 according to the group;

[0050] Step 2: Preparation of latex reagent R2: Dilute polystyrene microspheres with a particle size of 60nm to 130nm and 190nm to 220nm with the second buffer solution to a solution with a mass concentration of 0.5%-2%, and add 2.5mg / mL Carbodiimide hydrochloride and 5 mg / mL N-hydroxysuccinimide were reacted at room temperature for 2 hours, the supernatant was removed by centrifugation, and the precipitate was diluted with the second buffer to a mass concentration of polystyrene microspheres of 0.5 %, repeat the centrifugation above, remove the supernatant, and dilute the precipitate with the second MES buffer until the mass concentration of polystyrene microspheres is 1%-2%, then add anti-human serum amyloid A1 antibody and anti-human Serum amyloid A2 antibody until the mass conc...

Embodiment 1

[0059] Prepare hemolytic agent R1 according to the following component contents:

[0060]

[0061] Take 1L of purified water, add the above-mentioned raw materials in sequence, stir to dissolve, filter and pack to obtain the hemolytic agent R1.

[0062] Preparation of latex reagent R2: Dilute polystyrene microspheres with particle diameters of 100nm and 220nm with 50mmol / L MES buffer respectively to a solution with a mass concentration of 1%, add 2.5mg / mL carbodiimide hydrochloride Salt, 5mg / mL N-hydroxysuccinimide, react at room temperature for 2 hours, use a high-speed centrifuge, centrifuge 220nm latex at 9000rpm / min for 1 hour, and 100nm latex at 12000rpm / min Centrifuge for 2 hours, remove the supernatant, dilute the precipitate with 50mmol / L MES buffer to a mass concentration of polystyrene microspheres of 0.5%, repeat the above centrifugation, remove the supernatant, and buffer the precipitate with 50mmol / L MES solution to dilute the mass concentration of polystyrene...

Embodiment 2

[0064] Prepare hemolytic agent R1 according to the following component contents:

[0065]

[0066] Take 1L of purified water, add the above-mentioned raw materials in sequence, stir to dissolve, filter and pack to obtain the hemolytic agent R1.

[0067] Preparation of latex reagent R2: Dilute polystyrene microspheres with a particle size of 90nm and 220nm with 50mmol / L MOPS buffer respectively to a solution with a mass concentration of 2%, and add 2.5mg / mL carbodiimide hydrochloride Salt, 5mg / mL N-hydroxysuccinimide, react at room temperature for 2 hours, use a high-speed centrifuge, centrifuge 220nm latex at 9000rpm / min for 1 hour, and 90nm latex at 12000rpm / min Centrifuge for 2 hours, remove the supernatant, dilute the precipitate with 50mmol / L MOPS buffer to a polystyrene microsphere mass concentration of 0.5%, repeat the above centrifugation, remove the supernatant, and then dilute the precipitate with 50mmol / L MOPS buffer until the mass concentration of polystyrene mi...

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Abstract

The invention provides a kit for detecting serum amyloid A content and a preparation method and detection method thereof and belongs to the field of medical detection. The kit comprises a hemolytic agent R1 and a latex reagent R2 which are independent with each other. The latex reagent R2 comprises latex coated with a serum amyloid A antibody. The latex coated with the serum amyloid A antibody isprepared from latex having particle sizes of 60-130nm and coated with the anti-serum amyloid A1 antibody and latex having particle sizes of 190-220nm and coated with the anti-serum amyloid A2 antibodythrough mixing. The invention also provides a preparation method and a detection method of the kit for detecting serum amyloid A content. The kit can detect whole blood, serum, a cerebrospinal fluidand a pleuroperitoneal fluid, utilizes a small amount of samples, is free of separation or pretreatment, can be directly used for detection and has good sample compatibility.

Description

technical field [0001] The invention belongs to the field of medical detection, and relates to a kit, a preparation method and a detection method for measuring serum amyloid A content. Background technique [0002] Serum amyloid A protein (SAA) is an acute phase response protein secreted by the liver. It comes from the apolipoprotein family and is the precursor of tissue amyloid A. Its plasma concentration increases significantly under conditions such as inflammation, trauma, myocardial infarction, infection, and tumor, and can be used as a chemokine for immune cells such as lymphocytes; it can also up-regulate various inflammatory factors (such as TNF, IL-6, IL-8, etc. )expression. SAA increases significantly during virus infection, and increases 4-6 hours after infection, and disappears faster during the recovery period. The increase in patients with acute infection reflects the severity of the disease. For some diseases, such as viral infection, transplant rejection, c...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N21/31G01N21/82
CPCG01N33/68G01N21/31G01N21/82G01N2333/47
Inventor 蔡清华何浩会赵亚荣孙成艳
Owner DIRUI MEDICAL TECH CO LTD
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