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Detection method of hemorrhagic fever with renal syndrome IgM antibodies and reagent kit

A technology for hemorrhagic fever virus and renal syndrome, applied in the field of biological detection, can solve the problems of susceptibility to external temperature, short storage period of gold standard solution, precipitation phenomenon, etc., to improve sensitivity, avoid gold standard substance aggregation and cross-reaction little effect

Active Publication Date: 2015-02-11
山东康华生物医疗科技股份有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide a method for detecting IgM antibody of hemorrhagic fever with renal syndrome virus and a test kit suitable for clinical auxiliary diagnosis and differentiation, so as to solve the problem that the gold standard solution in the prior art has a short shelf life and is easy to detect. Affected by external temperature and other factors, precipitation and other problems occur

Method used

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  • Detection method of hemorrhagic fever with renal syndrome IgM antibodies and reagent kit
  • Detection method of hemorrhagic fever with renal syndrome IgM antibodies and reagent kit
  • Detection method of hemorrhagic fever with renal syndrome IgM antibodies and reagent kit

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Effect test

Embodiment 1

[0026] Preparation of gold standard working solution

[0027] 1. Raw materials: chloroauric acid, trisodium citrate, mouse anti-human IgM monoclonal antibody, hydrochloric acid, potassium carbonate, tris, trehalose, sodium chloride, glycerin, bovine serum albumin, PROCLIN300, salicylic acid Isooctyl Acetate, Xanthan Gum, PEG20000, Carboxymethyl Chitosan.

[0028] 2. Instruments and equipment: volumetric flask; magnetic stirrer; 1 / 10,000 electronic balance; refrigerator; centrifuge.

[0029] 3. Preparation steps:

[0030] (1) Preparation of Gold Standard Concentrate: Take 100ml of 1wt% chloroauric acid aqueous solution, heat to boiling, add 0.8ml of 0.5wt% trisodium citrate solution, and mix quickly until the color of the solution changes from light yellow to blue to purple To completely transparent wine red, continue to boil for 10 minutes, stop heating, and cool to room temperature;

[0031] Add 0.3 ml of mouse anti-human IgM monoclonal antibody of 1 mg / ml to the obtained ...

Embodiment 2

[0049] Preparation of washing solution

[0050] 1. Raw materials: Tris, sodium chloride, sodium hydroxide, propanol, sodium dihydrogen phosphate, disodium hydrogen phosphate, bovine serum albumin, PROCLIN300, dichloroacetic acid, TritonX-100, ethylenediamine Tetraacetic acid EDTA, PEG20000.

[0051] 2. Instruments and equipment: volumetric flask; magnetic stirrer; 1 / 10,000 electronic balance; refrigerator.

[0052] 3. Preparation steps:

[0053] Put 121.1 mg of Tris, 0.1 mg of sodium chloride and 0.1 mg of bovine serum albumin in a volumetric flask, add double distilled water, shake gently to dissolve completely, then continue to add double distilled water until 100mL, add 0.15% EDTA, then add propanol with a volume ratio of 1.5%, add 1.2% dichloroacetic acid and 0.3% TritonX-100 after mixing, and finally adjust its pH with 0.1M dilute sodium hydroxide Adjust to 8.6 to obtain the washing liquid, put it in the washing liquid bottle, and store it at 2-8°C;

[0054] Comparati...

Embodiment 3

[0062] A detection method for the IgM antibody of the hemorrhagic fever with renal syndrome virus. The antigen of the hemorrhagic fever with renal syndrome virus is fixed on the nitrocellulose membrane, and the serum to be tested is added. Antigen binding on the membrane, then add gold standard working solution containing mouse anti-human IgM monoclonal antibody, mouse anti-human IgM monoclonal antibody combined with renal syndrome hemorrhagic fever virus IgM antibody in the serum to be tested, showing red color, and finally add washing solution to wash away excess gold standard working solution and other impurities;

[0063] Gold standard working solution is prepared by the following method:

[0064] ⑴Preparation of Gold Standard Concentrate: Take 100ml of 0.5wt% chloroauric acid aqueous solution, heat to boiling, add 0.8ml of 0.4wt% trisodium citrate solution, mix well, and wait until the color of the solution changes from light yellow to blue to purple to complete Transpar...

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Abstract

The invention relates to the technical field of biological detection, in particular to a detection method of hemorrhagic fever with renal syndrome IgM (immunoglobulin M) antibodies and a reagent kit. The detection method comprises the steps that hemorrhagic fever with renal syndrome antigens are fixed on a membrane; to-be-detected serum is added; the hemorrhagic fever with renal syndrome IgM antibodies in the to-be-detected serum are combined with the antigens fixed on the membrane; gold labeled operating fluid containing mouse anti-human IgM monoclonal antibodies is added; the mouse anti-human IgM monoclonal antibodies are combinec with the hemorrhagic fever with renal syndrome IgM antibodies in the to-be-detected serum; a red color is shown; scrubbing liquid is added finally; and the redundant gold labeled operating fluid and other impurities are washed out. The reagent kit has the characteristics of longer quality guarantee period, less cross reaction, better color rendering performance and the like.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to a detection method and kit for hemorrhagic fever with renal syndrome virus IgM antibody. Background technique [0002] Hemorrhagic fever with renal syndrome (Hemorrhagic fever with renal syndrome, hereinafter referred to as HFRS), commonly known as Epidemic hemorrhagic fever (EHF) in China, is caused by Hantavirus (HV). Damage is the main clinical feature of natural foci of infectious diseases, rats are the main source of infection, parasitic mites participate in the transmission, mainly through contact with host animal excreta through wounds, aerosols, and diet. Its clinical manifestations are complicated and its prognosis is poor. [0003] The laboratory diagnostic methods for HFRS mainly include routine examination, ultrasonography, genetic testing, virus isolation and identification, and serological testing. At present, the clinical diagnosis is mainly based on...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/569G01N33/532
Inventor 杨致亭杨金红王秀利武新清孙洪芝刘迎军袁忠群王德亚
Owner 山东康华生物医疗科技股份有限公司
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