抗P1抗体检测的血清学分析和方法探讨

Serological analysis and method discussion of anti-P1 antibody detection

目的:分析P1(-)表型个体产生抗P1抗体的血型、抗体鉴定的血清学检测结果、分子机制以及交叉配血策略,并回顾对比分析既往2例血清学检测方法和结果,对优化检测方法提高抗P1抗体检出率进行探讨。方法:采用盐水法、聚凝胺和微柱凝胶法进行血型鉴定、抗体筛选和交叉配血,采用PCR扩增α-1,4-半乳糖基转移酶基因(α-1,4-galactosyltransferase,A4GALT)和β-1,3-N-乙酰半乳糖氨基转移酶基因(β-13-N-acetylgalactosaminyltransferase,B3GALNT1)外显子编码区,分析DNA序列。结果:患者1经过盐水法、微柱凝胶法、抗人球蛋白法血清学分析鉴定为IgM抗P1抗体,经过PCR核酸检测确定为患者表型为P1(-);P型,验证与血清学的检测结果一致。患者2仅通过血清学检测鉴定为抗P1抗体。结论:抗P1抗体多为IgM型抗体,在进行抗体检测时需考虑到检测方法的适用范围和局限性,选择合适的方法,并进行多种方法联合检测相互验证的方式,以提高抗体的检出率。核酸检测基因分型技术的应用可以从分子生物学角度提供重要佐证,有助于提高抗体鉴定的准确率。

Objective:To review the blood type of anti-P1 antibody produced by individuals with P1(-)phenotype,the serological test results of antibody identification,molecular mechanism and cross-matching strategy,compare the previous two serological test methods and results,and discuss the optimization of test methods to improve the detection rate of anti-P1 antibody.Methods:Blood group identification,antibody screening and cross matching were performed by normal saline method,polybrene and microcolumn gel method,and PCR amplification was usedα-1,4-galactosyltransferase gene(A4GALT)andβ-1,3-N-acetylgalactosamine aminotransferase gene(B3GALNT1)exon coding region,and DNA sequence was analyzed.Results:Patient 1 was identified as IgM anti-P1 antibody by normal saline method,microcolumn gel method and antiglobulin method,and the phenotype of patient 1 was P1(-)by PCR nucleic acid detection.P type,was verified to be consistent with the results of serological detection.Patient 2 was identified with anti-P1 antibodies only by serologic testing.Conclusion:Most of the anti-P1 antibodies were IgM.When performing antibody detection,it is necessary to consider the scope and limitations of the detection methods,select the appropriate method,and conduct a variety of combined detection methods to verify each other,so as to improve the detection rate of antibody.The application of nucleic acid detection genotyping technology can provide important corroboration from a molecular biology perspective and help to improve the accuracy rate of antibody identification.