Development of a rapid and precise method of identifications for blood groups with weak antigens

• Main Authors: Nain-En Lu, 呂念恩
• Other Authors: 楊台鴻
• Format: Others
• Language: en_US
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/88280144952937761275

碩士 === 國立臺灣大學 === 醫學工程學研究所 === 98 === There is a problem that the false negatives of red blood cells(RBCs) with Rh(Del), causing hemolytic disease during blood transfusion and hemolytic disease of newborn (HDN), is sometimes happened in our blood centers where medical technologists identify them in general blood typing cards (Ortho BioVue System Microtyping card). Adsorption and elution method, the standard procedure of identifying Del, spends more times (at least an hour), quantities of sample (1c.c. RBCs), and costs (1c.c. reagent) than the method of microtyping card so that they do not use this method at pretransfusion testing. To solve this problem, we suppose that the appearance of false negative is due to less D antigens per RBC membrane than D positive persons. RBCs have no probability to agglutinate by monoclonal anti-(RhD) antibody. If we amplify D antigens from RBCs with Del, they could have been agglutinated because of the probability of agglutination arising. Base on this conception, we test four different kinds of antigen-antibody amplification techniques: the first is that a water soluble polymer, poly(acrylic acid), conjugated with secondary antibody, anti-(mouse IgG) which binds on anti-(RhD) antibody, would be used as an cross-linker that let RBCs with Del agglutinate; the second is that a new polymer (Ac-pAAm -Biotin), made from a water soluble polymer, poly(allylamine) which was partially acetylation and biotinylation, would be interact with anti-(RhD), anti-(mouse IgG)-biotin conjugates, and avidin to agglutinate RBCs with Del. The third and fourth are that alternatives of the new polymer (pAAm-Ac-Biotin), commercial bovine serum albumin, biotin conjugates (BSA-biotin) and poly(acrylic acid) beads, biotin conjugates (pAA beads-biotin), would be applied to agglutination of RBCs with Del. Our result revealed that only the third method, BSA-biotin, can prevent false negative efficaciously and this method is more rapid (about 10 mins) and less quantities of sample (10λ, 3% RBCs) and costs (20λ reagent) than adsorption and elution method.