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Protein suspension chip for synchronously detecting various antibodies in serum sample and preparation method and using method thereof

A technology of simultaneous detection and levitating chips, applied in the fields of resisting vector-borne diseases, measuring devices, instruments, etc., can solve the problems of lack of models and evaluation, and achieve good detection results.

Inactive Publication Date: 2011-01-05
CHINESE ACAD OF INSPECTION & QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, whether the suspension chip method can simultaneously detect West Nile antibody, Tula antibody, dengue fever antibody, and avian influenza antibody in human serum, and its quantitative detection ability, there is still a lack of models and evaluations.

Method used

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  • Protein suspension chip for synchronously detecting various antibodies in serum sample and preparation method and using method thereof
  • Protein suspension chip for synchronously detecting various antibodies in serum sample and preparation method and using method thereof
  • Protein suspension chip for synchronously detecting various antibodies in serum sample and preparation method and using method thereof

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Experimental program
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Effect test

preparation example Construction

[0049] 2. Preparation of samples to be tested

[0050] 1. Preparation of target analyte samples

[0051] The target analytes were rabbit anti-Tura IgG and rabbit anti-dengue type 2 antibody. The stock solution concentration of rabbit anti-Tura IgG was 44.2 μg / mL, and the stock solution concentration of rabbit anti-dengue type 2 antibody was 1.06 mg / mL.

[0052] Dilute the rabbit anti-Tura IgG, rabbit anti-Dengue type 2 antibody to be analyzed with the sample diluent in a 4-fold ratio serially to samples of different concentrations, so as to draw a standard curve of sample detection dose-response, and several of the samples have low concentrations For detection sensitivity, high concentration samples should saturate the binding sites of the encoded microspheres.

[0053] 2. Processing of human serum samples

[0054] The human serum sample is diluted 1:10 with the sample dilution, for example, 5 μL of the human serum sample plus 50 μL of the sample diluent is mixed and then u...

Embodiment 1

[0055] Embodiment 1, the preparation of the protein suspension chip that detects several virus antibodies

[0056] 1. Capturing antigen-coated encoded microspheres

[0057] The No. 025, No. 028, No. 031, and No. 032 coded microspheres used in the present invention were purchased from Bio-Rad Company of the United States. The No. 025 coded microspheres are used to label the West Nile E protein antigen that can capture West Nile antibodies. West Nile E protein-coated microspheres; coded microspheres No. 028 are used to label Tula fopA protein antigens that can capture Tula antibodies, that is, microspheres coated with Tula fopA protein; coded microspheres No. 031 are used to label Tura fopA protein antigens that can capture Tula antibodies The dengue E2 protein antigen that captures the dengue antibody, that is, the dengue E2 protein is used to coat the microsphere; the 032 coded microsphere is used to label the avian influenza H5 protein antigen that can capture the avian influ...

Embodiment 2

[0069] Embodiment 2, optimization of suspension chip preparation method conditions

[0070] 1. Selection of the amount of antigen coating on microspheres

[0071] 100 μL of encoded microspheres were coated with 1 μg, 2.5 μg, 5 μg, 7.5 μg, 10 μg, 12.5 μg, 15 μg, 20 μg, 25 μg, 30 μg, 40 μg, 50 μg, respectively. After testing the effect comparison, with 1~50μg / 1.25×10 6 Microspheres, that is, 4-200ng / 5000 microspheres / test, the coating effect is the best, counted under the microscope and stored in dark and refrigerated until use. Such as figure 1 As shown, No. 025 microspheres coated with West Nile E protein, No. 028 microspheres coated with Tula fopA protein antigen, No. 031 encoded microspheres of dengue E2 protein, and No. 032 encoded microspheres of avian influenza H5 protein all fell on the It is in the correct detection area and obtains high signal-to-noise ratio results (MFI value is much greater than 2000), indicating that the optimized suspension chip detection system...

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Abstract

The invention discloses a protein suspension chip for synchronously detecting various antibodies in serum sample and a preparation method and a using method thereof. The method has the advantages of good detection capacity, high sensitivity, strong specificity and wide dynamic range, and an open detection modular platform of the protein suspension chip for detecting microbial antibodies represented by West Nile antibody, Francisella tularensis antibody, dengue fever antibody and influenza antibody is established.

Description

technical field [0001] The invention relates to a protein suspension chip for synchronous detection of West Nile antibody, Tula antibody, dengue fever antibody and avian influenza antibody in serum samples, a preparation method and a use method thereof. Background technique [0002] West Nile fever is an acute infectious disease caused by West Nile virus infection. The majority of human infections with West Nile virus manifest as recessive infection, and there is 1 clinical case in about 140 to 300 infected persons. Symptoms include sudden fever, headache, back pain, and muscle pain. Fever can be manifested as double-wave fever, and about half of the patients can see a rash. The rash appears during the febrile period or at the end of the febrile period. The rash can be roseate or maculopapular, mainly on the back of the neck and upper limbs, and lasts for about 1 week. Visible pharyngitis and nausea, vomiting, diarrhea, abdominal pain and other gastrointestinal symptoms....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N21/64
CPCY02A50/30
Inventor 王静杨永莉杨宇孙肖红曹晓梅张晓龙
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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