Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Platelet anti-human globulin experimental immune zone distribution detection method

A technology for detecting the distribution of anti-human globulin, applied in the field of immunology and medical testing, can solve the problems of complicated experimental procedures, easy to be activated and aggregated, and long operation time, so as to improve accuracy and sensitivity, avoid loss of antigenicity, The effect of high detection accuracy

Pending Publication Date: 2021-02-19
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI +1
View PDF10 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the gold standard method of platelet antibody analysis in the world, "The monoclonal antibody immobilization of platelet antigen assay (MAIPA)" has complicated experimental procedures, and the operation time is as long as 5 hours or more. High, cannot be used in clinical routine testing applications
[0005] The anti-human globulin test is a classic method for detecting incomplete antibodies of red blood cells, but it has not been effectively applied to the detection of platelet antibodies. The main reason is that the red blood cells are visible red to the naked eye, regular in shape, relatively uniform in size, and easy to determine agglutination and non-agglutination
However, the platelet itself has no color, irregular shape, uneven size distribution, and is easy to be activated and aggregated, so it is difficult to detect antibodies by agglutination method

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
  • Platelet anti-human globulin experimental immune zone distribution detection method
  • Platelet anti-human globulin experimental immune zone distribution detection method
  • Platelet anti-human globulin experimental immune zone distribution detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1: Preparation of quantum dot immunolabeled anti-human globulin

[0064] (1) Preparation of anti-human globulin

[0065] Extract gamma globulin from human plasma, emulsify with adjuvant, immunize sheep (1 mg / rat) or rabbit (0.5 mg / rat), etc., immunize at multiple points each time, and draw venous blood after 5 times of immunization. The whole blood was centrifuged at 2000rpm for 30min to separate the plasma, and the protein was precipitated with 50% ammonium sulfate, and purified with DEAE-52 ion exchange chromatography column, the purified protein was collected, and the protein concentration was determined by SDS-PAGE electrophoresis purity test and test tube anti-globulin After the titer is detected by the protein experiment, it is ready for use. Test results such as figure 2 Shown, wherein, 1, Marker; 2, 0.05M NaCl solution eluent; 3, 0.1M NaCl solution eluent. The test results showed that the purity of the antihuman globulin in the eluate of 0.05M NaCl s...

Embodiment 2

[0071] Example 2: Platelet Antibody Detection

[0072] (1) Preparation of platelet suspension

[0073] Three portions of fresh O-type EDTA anticoagulated whole blood were collected and centrifuged at 900 rpm for 10 minutes to obtain platelet-rich plasma, which was diluted 2-fold with PBS solution containing 1% BSA to form a platelet suspension. If platelets are collected by machine, the platelet suspension is diluted 10 times with PBS solution containing 1% BSA.

[0074] (2) Preparation of zone distribution detection card

[0075] Add 100 μL of immune complex separation enhancement medium into the reaction chamber of the zone distribution detection card. The immune complex separation enhancement medium has a specific gravity of 1.05 to 1.10, an osmotic pressure of 280 to 350 mmol / L, and contains 0.1 to 1 mg / mL of secondary antibody. The solution was centrifuged at 1000rpm for 1min before use.

[0076] (3) Detection steps

[0077] Add 50 μL platelet suspension, 50 μL sample...

Embodiment 3

[0081] Embodiment 3: platelet cross-matching type

[0082] (1) Preparation of platelet suspension

[0083] Collect EDTA anticoagulated whole blood from blood donors with the same ABO blood type as the patient, centrifuge at 900 rpm for 10 minutes, and take the upper layer of platelet-rich plasma (PRP); dilute it with PBS solution containing 1% BSA twice to form a platelet suspension. If platelets are collected by machine, the platelet suspension is diluted 10 times with PBS solution containing 1% BSA.

[0084] (2) Preparation of test card

[0085]Add 100 μL of immune complex separation enhancement medium to the reaction chamber of the zone distribution detection card. The immune complex separation enhancement medium has a specific gravity of 1.05-1.10, an osmotic pressure of 280-350 mmol / L, and contains 0.1-1 mg / mL secondary antibody solution , centrifuged at 1000rpm for 1min and then used for later use.

[0086] (3) Cross-matching step

[0087] Add 50 μL donor platelet su...

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
Osmotic pressureaaaaaaaaaa
Thicknessaaaaaaaaaa
Login to View More

Abstract

The invention discloses a platelet anti-human globulin experiment immune zone distribution detection method, which adopts an immunolabelled anti-human globulin experiment, and uses immunolabellers including but not limited to radioactive isotopes, colloidal gold, fluorescein, quantum dots, chemical (or biological) luminescent agent indications or organic dyes to label anti-human globulin reagents.Reaction and combination with a platelet-specific antibody are then carried out, and centrifugally entering the zone distribution detection card to present different zone distributions is conducted,thereby judging the detection result. The method disclosed by the invention is simple to operate, intuitive and reliable in result, low in cost and accurate in result, can meet the requirements of clinical conventional platelet antibody detection, assists the diagnosis of clinical platelet-related immune diseases and the compatibility experiment detection before blood transfusion, reduces the occurrence of platelet infusion invalidation and bleeding and death cases caused by the platelet infusion invalidation, and clinical safe, effective and scientific platelet blood transfusion is guaranteed, and meanwhile precious platelet resources can be saved.

Description

technical field [0001] The invention relates to the fields of immunology and medical examination, in particular to a method for detecting the distribution of platelet antihuman globulin experimental immune zones. Background technique [0002] There are complex blood group antigens on the surface of platelets, including ABO antigens, HLA-I antigens, platelet-specific HPA antigens, and CD36 antigens, all of which can stimulate the body to produce platelet antibodies. Platelet antibodies can cause platelet immune damage in patients, resulting in immune thrombocytopenia, such as ineffective platelet transfusion, post-transfusion purpura, autoimmune thrombocytopenia, and neonatal thrombocytopenia. In particular, anti-platelet CD36-specific antibodies are of great significance in the occurrence of clinical platelet immune diseases in my country. [0003] Platelet transfusion is currently one of the most important treatments for thrombocytopenia and various diseases. A large numb...

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
IPC IPC(8): G01N33/68
CPCG01N33/6893G01N2800/222Y02A50/30
Inventor 段生宝李勇陈维佳冯立岗丁少华王红梅谢劲松王玉珏陈晔洲魏双施田晶晶刘杰王泽龙刘永茂隋金晶冷向武张妍
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com