ABO血型反定型梯形微板法的建立

目的建立梯形微板ABO血型反定型检测方法。方法随机选用常规A、B、O、AB型血样各8份。运用正交设计原理选择血浆稀释度、红细胞浓度、反应温度、静止时间4个参数的最佳条件;采用混合血样倍比稀释,确定凝集与非凝集灰区值。根据已确定的最佳条件和灰区值,进行重复性试验、灵敏度试验、准确性试验、脂血干扰试验、溶血干扰试验,并检测6份ABO亚型血样AM、AX、A2B、A3B、AXB、BM和5份含不规则抗体血样抗-M、抗-N、抗-P1、抗-Leb、抗-HI,以评价对特殊血样的检出能力。结果重复性符合率100%。灵敏度可达到与试管法相同效价的检出能力。准确性达99.99%。47份不同脂血程度血样抗干扰试验一次判读正确率100%。溶血干扰试验在Hb≤20g/L时,对梯形微板法试验结果无明显影响,在严重溶血情况下(Hb>20g/L),对凝集端无影响,对非凝集端有影响。对11份特殊血样均有较好的检出能力。结论梯形微板法不仅能够很好地完成普通血样ABO反定型的检测,还具有提高对弱凝集血样检测灵敏度的优势,进而提高试验的准确性。

Establishing a trapezoid micro-plate method to detect ABO blood group in serum

Objective Establishing a trapezoid micro-plate method to detect ABO blood group in serum.Methods Select 32 blood samples containing A,B,O or AB types randomly.Use orthogonal design principle to select the optimum conditions of the blood plasma dilution,erythrocytes density,temperature and static time.The mix-blood samples are serially diluted.The agglutinating and nonagglutinating gray zones are also defined.Based on the optimum conditions and criterions,the reproducibility test,sensitiveness test,accuracy test,lipaemia interference test and haemolysis interference test are carried out.6 ABO subgroup(AM,AX,A2B,A3B,AXB,BM) and 5 irregular antibody(anti-M,anti-N,anti-P1,anti-Leb,anti-HI) are detected to evaluate the ability of the trapezoid micro-plate method to detect special samples.Results The trapezoid micro-plate method's repetitiveness tests coincidence is 100%.Its sensitivity is identical to the test tube method.The accuracy is 99.99%.In the 47 various kinds of lipaemia blood samples' anti-interference tests,the correct interpretation at one time is 100%.In the haemolysis interference tests,when hemoglobin content are below 20g/L,there are no obvious influences found.When sever haemolysis appears(blood plasma hemoglobin content are above 20g/L),there is no effect on the agglutination,but make a difference to the nonagglutinating.The ability to dectect 6 ABO blood subgroups and 5 irregular antibody blood samples is great.Conclusion The trapezoid micro-plate method not only has good ability to detect the ABO blood group both in the red cell typing and serum typing,but also enhances the accuracy in detecting the agglutinating blood samples and improves the exactitude of the tests.