Novel Miltenberger blood group antibody detecting method

Abstract

The invention discloses a non-blood red cell dependent type Miltenberger blood group antibody detecting method. The detecting method comprises the following steps of: coupling sheep red blood cell subjected to hydroformylation and magnetization with polypeptide antigen containing a connecting chain to form an artificial screening cell; and uniformly mixing the artificial screening cell with a detecting sample for cultivating and washing, and measuring the result after adding fluorescence-labeled secondary antibody for cultivating and washing. By virtue of the way, the non-blood red cell dependent type Miltenberger blood group antibody detecting method provided by the invention is free of centrifuging for the washing due to the application of magnetic force, simple and convenient to operate, capable of realizing simultaneously detecting complete antibody and incomplete antibody, short in detecting process time consumption, easy to automate, accurate in detected result and capable of realizing semi-quantitative or quantitative detection by combining with a detecting apparatus, and the result is easy to judge. Besides, the detecting method can guarantee safe, efficient and scientific blood transfusion in clinic.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a new method for detecting Miltenberger blood group antibodies, that is, a method for detecting red blood cell-independent Miltenberger blood group antibodies. Background technique [0002] The MNS blood group system is the second blood group system discovered after ABO, and the number of polymorphisms in its antigens is second only to the Rh blood group system. In the MNS blood group system, there is a special Miltenberger subblood group system, whose blood group antigens are erythrocyte phenotypes combined with a series of low-frequency antigens identified by human serum immune antibodies. The occurrence frequency of several antigens such as Mur and Mil in this subsystem is less than 1 / 1000 in European Caucasians, but it is 3% to 10% in Chinese and other Asian populations. A survey of 844 Dong people in Guangxi showed that the positive rate of the Mur rare blood group antigen was ...

AI Technical Summary

Problems solved by technology

Due to the lack of relevant reagents, the antibody screening cells used by most medical institutions do not contain Miltenberger blood group antigen, and routine testing cannot identify the specificity of the antibody, resulting in missed detection of patients with this antibody

Due to the short storage time of fresh red blood cells, up to 3 months, the sources of Miltenberger antigen-positive red blood cells are very limited, and the detection of Miltenberger blood group screening and its antibodies is greatly limited

At present, the main problems faced in the detection of Miltenberger blood group antibodies are: 1) Because the corresponding screened red blood cells are not used in the clinical test spectrum cells, irregular antibodies are often missed; 2) The stable supply of screened red blood cells containing rare antigens cannot be obtained for a long time Guarantee; 3) The storage time of fresh red blood cells is extremely short

[0004] Since there is no commercially available anti-Miltenberger blood group antigen monoclonal antibody, the inherent shortcomings of human blood group reagents (human erythrocyte reagents and human serum antibody reagents) and the lack of suitable test methods, it is currently not possible to routinely conduct clinical trials in various countries around the world. Carry out Miltenberger blood type compatibility test, only in a small number of large blood centers and blood type reference laboratories for some difficult blood type incompatibility disease patients

So far, the specific obstacles and problems that cannot routinely carry out Miltenberger blood group compatibility test in clinical practice are: the shortage of both Miltenberger antigen reagents and antibody reagents, the inherent shortcomings of red blood cell reagents and human serum antibody reagents, and the currently applied immunoserological test technology disadvantages of the method, etc.

But CSL companies use Kode TM The blood group antigen reagents prepared by the technology are still based on surviving red blood cells. The above-mentioned fundamental problems have not been solved. The blood group antigens on the membranes of surviving red blood cells are easily hemolyzed, and the antigenicity also disappears.

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