Joint detection device and preparation method of helicobacter pylori urease antibodies IgM and IgG
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A combined detection technology for Helicobacter pylori, applied in measuring devices, instruments, disease diagnosis, etc., can solve problems such as inability to classify antibodies, and achieve the effects of simple operation, cost saving, and improved sensitivity
Inactive Publication Date: 2015-07-01
吉林双正医疗科技有限公司
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The traditional detection method of Helicobacter pylori urease antibody can only det
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[0033] The preparation method of the Helicobacter pylori urease antibody IgM, IgG combined detection device of the present invention, comprises solid phase highly specific mouse anti-human IgM, IgG antibody (detection line T1, T2) and goat anti-mouse IgG multiplex with purification The nitrocellulose membrane of the cloned antibody (control line), the glass fiber adsorbed with colloidal gold-labeled Helicobacter pylori urease antigen (HP Ure-Ag) and mouse IgG, sample pad, absorbent paper and other auxiliary materials were pasted together. The specific steps are as follows:
[0034] (a) Preparation of colloidal gold by trisodium citrate reduction method;
[0035] (b) using the colloidal gold prepared in step (a) to label Helicobacter pylori urease B antigen to obtain immune colloidal gold;
[0036] (c) Diluting the immune colloidal gold in step (b) with a gold spraying buffer to obtain an immune colloidal gold solution, and spraying the immune colloidal gold solution on the gl...
Embodiment
[0044] Preparation of Helicobacter Pylori Urease Colloidal Gold
[0045] 1. Preparation of colloidal gold solution
[0046] Quickly add gold chloride into heated 100ml purified water, and after the solution boils again, quickly add trisodium citrate, gold chloride: trisodium citrate 1: (0.5-2), continue to boil, and observe that the color of the solution changes from yellow to After the black turns purple again, and finally turns into a stable wine red, continue heating for 10 minutes by timing. The colloidal gold particles are 20-60nm, and 30nm colloidal gold is selected for the next experiment.
[0047] 2. Preparation of immunocolloidal gold
[0048] 1) Take 100ml of 30nm colloidal gold solution, add 140μl of pH regulator, mix well; let stand for 5min;
[0049] 2) In the 30nm colloidal gold solution, add Helicobacter pylori urease at a ratio of 14 μg per milliliter of colloidal gold solution, a total of 1.4 mg, mix well; let stand for 5 minutes;
[0050] 3) Add 0.4 ml...
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Abstract
The invention relates to a joint detection device and a preparation method of helicobacter pylori urease antibodies IgM and IgG. The helicobacter pylori urease antibodies are prepared from a nitrocellulose membrane, a glass fiber adsorbing a colloidal gold labeled helicobacter pylori urease antigen and a mouse IgG, a sample pad, absorbent paper and other auxiliary materials, wherein the materials are adhered together, and the nitrocellulose membrane contains a purified high-specificity mouse-anti-human IgM and IgG antibodies, and a goat-anti-mouse antibody in a solid phase manner. The joint detection device has the advantages that the structure is simple, the conception is novel, the nitrocellulose membrane are coated with the anti-human IgM and IgG antibodies, so that the specificity is strong, and the helicobacter pylori urease antibodies IgM and IgG in a specimen are simultaneously detected without increasing the production operation complexity. Proper gold spray buffer and sample pad treating fluid are matched to effectively improve the reaction sensitivity on the basis of guaranteeing complete release of immunocolloidal gold, and under the same threshold, the use amount of the immunocolloidal gold can be reduced to save the cost. The detection device is high in sensitivity, strong in specificity, simple, convenient and strong in practicability and can realize the time-saving aim during operation.
Description
technical field [0001] The invention relates to the field of medical detection equipment, in particular to a combined detection device and preparation method for Helicobacter pylori urease antibody IgM and IgG, which can realize sensitive, specific and rapid detection of Helicobacter pylori urease antibody IgM and IgG. Background technique [0002] Helicobacter pylori (Hp) infection plays an important role in the pathogenesis of gastrointestinal diseases such as chronic gastritis, peptic ulcer, and gastric cancer. Epidemiological surveys have shown that the infection rate of HP among adults in my country is above 50%. In the detection of Helicobacter pylori infection, bacterial culture, histopathological examination, urease test, breath test and other methods may cause invasive pain to patients, or require certain conditions and techniques, or because special instruments are required, Or because of high cost, etc., its clinical promotion and repeated follow-up review are li...
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