Heparin binding protein detection test paper for immunomicrosphere chromatography detection

The technology of heparin-binding protein and immune microspheres is applied in the field of preparation of test strips for detecting heparin-binding protein by immunomicrosphere chromatography, which can solve the problems of no diagnostic reagent method, low accuracy and stability, long time consumption, etc. Achieve the effect of ensuring validity and quality control, clear and accurate test results, and strong repeatability

Inactive Publication Date: 2018-07-10
PRO MED BEIJING TECH
View PDF3 Cites 15 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, the HBP detection reagents used in western countries are mainly ELISA method, which is very popular in clinical research abroad, but due to the need for professional testing personnel, the operation is complicated and time-consuming, so it cannot be tested at the bedside
At the same time, Lufthansa Pharmaceuticals of Sweden patented the application of HBP as a biomarker for the diagnosis of sepsis, and patented HBP levels as a diagnostic method for urinary tract infection and a diagnostic method for bacterial meningitis, but the company did not have a clear diagnostic reagent method
Axis-Shield registered the heparin-binding protein ELISA method kit in CFDA, but because the ELISA method detects heparin-binding protein, the clinical operation is complicated, time-consuming, and clinically not popularized. CN204882574U discloses a heparin-binding protein quantitative detection kit, which The patent uses colloidal gold immunochromatography. Due to the limitations of accuracy and stability of colloidal gold immunochromatography, the market urgently needs a more accurate and sensitive detection method for bedside heparin-binding protein detection.
Hangzhou Zhonghan Shengtai Biology and Henan Shengsheng Medical applied for patents CN204882575U and CN105572386A for the detection of heparin-binding protein by immunofluorescence chromatography, but neither of the patents uses a solid phase for fluorescently labeled antibodies, and the clinical operation is cumbersome. At the same time, the patent CN204882575U adds an auxiliary detection line , there is a hidden danger of unstable test results

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Heparin binding protein detection test paper for immunomicrosphere chromatography detection
  • Heparin binding protein detection test paper for immunomicrosphere chromatography detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Such as figure 1 Shown, a kind of immune microsphere chromatography detects the heparin-binding protein detection test paper preparation method, comprises the following steps:

[0040] S1. Specific steps of calibration curve preparation:

[0041] Specific steps: use normal human standard serum (purchased from Jiangsu Enmoase) to prepare 5ug / ml heparin-binding protein mother solution, dilute the mother solution heparin-binding protein standard product, and the heparin-binding protein dilution concentration is 0.1ug / L, 0.5ug / L, 1ug / L, 5ug / L, 10ug / L, 50ug / L, 100ug / L, 150ug / L, 200ug / L, 250ug / L, the signal value and concentration curve were calibrated with heparin-binding protein detection test paper and POCT immunoanalyzer.

Embodiment 2

[0043] S2. Preparation of detection buffer containing heparin-binding protein antibody

[0044] Specific steps: prepare rabbit anti-human heparin binding protein antibody (biocare) with PBS buffer solution of pH 7.4 and 10mM, the preparation concentration is 2ng / ul, add 5% sucrose, 5% BSA, 0.1% Tween-20 , 0.05% sodium azide.

Embodiment 3

[0046] S3. Preparation of microsphere-labeled secondary antibody

[0047] Specific steps: Take 100ml of 10% 0.3um fluorescent microspheres, wash with PH4.5 50mM MES solution for 3 times, add 1ml of freshly prepared 10mg / ml EDAC solution (prepared in PH4.550mM MES), incubate at room temperature for 30min , and then centrifuged at 2000rpm for 5min, then washed with PH4.550mM MES solution, incubated overnight at room temperature with 1ml of 1ug / ul chicken anti-rabbit secondary antibody (PBS), incubated with 10% BSA, 100×Tris-EDTA for 1 hour at room temperature , washed with 1ml PBS three times after centrifugation, stored in 4% sucrose, 0.05% sodium azide solution.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
particle diameteraaaaaaaaaa
Sensitivityaaaaaaaaaa
Login to view more

Abstract

The invention relates to the technical field of clinical infectious disease marker detection, in particular to heparin binding protein detection test paper for immunomicrosphere chromatography detection. A method includes the following steps that a calibration curve is prepared; a detection buffer solution containing rabbit anti-human heparin binding protein antibody is prepared, fluorescent or colorful latex microsphere-marked chicken anti-rabbit antibody is prepared, a sample pad coats a solid phase, a detection line coated with a mouse anti-human heparin binding protein antibody and a reaction pad coated with a goat anti-rabbit antibody quality control line are prepared, a separation membrane is assembled, a test paper strip is assembled, a clinical sample is detected, and the dried test paper strip is placed in an aluminum foil bag. The heparin binding protein detection test paper for fluorescence chromatography detection has high cost effectiveness, and the level of heparin binding protein in body fluids of patients is quantitatively detected and evaluated. The method is used for a POCT detection system and screening or detection of changes of heparin binding protein in the body fluids of the patients to determine a detection method of infectious disease markers so that the heparin binding protein detection technology can become a clinical conventional detection project.

Description

technical field [0001] The invention relates to the technical field of detection of heparin-binding protein, a biomarker of infectious diseases, and in particular to a method for preparing a test paper for detecting heparin-binding protein by immune microsphere chromatography. Background technique [0002] As an inflammatory marker, HBP has higher sensitivity and specificity than procalcitonin. Because HBP is mainly released by PMN by external stimuli, the HBP content in normal human blood is very low, generally not exceeding 10ng / mL. When an infection occurs, some bacteria invade the blood vessels, and the bacteria themselves or the toxins released by the bacteria Other substances stimulate neutrophils to release HBP, which leads to an increase in HBP content in the blood. HBP can reach 20-30 ng / mL in general infection, and severe infection in ICU may exceed 100 ng / mL or even as high as 1000 ng / mL; when the HBP content exceeds 1000 ng / mL, the patient is already in extreme ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/531
Inventor 范秋苹潘志红张宁高巍巍陈美艳南洋汪廷枫郝存田茹
Owner PRO MED BEIJING TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap