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目的对周口地区115例非亲缘关系的汉族无偿献血者ABH分泌型及非分泌型进行血清学和基因分析,初步探讨周口地区汉族人群ABH分泌型及非分泌型的FUT2基因多态性特点,以及PCR-SSP技术在ABO疑难血型鉴定应用的可行性。方法留取115例随机无偿献血者的唾液和外周血样本。唾液标本采用凝集抑制试验检测分泌型和非分泌型;采用试剂盒提取基因组DNA,设计A385T和G447A两对引物,采用聚合酶链反应-序列特异性引物(PCR-SSP)技术和测序技术,对115例健康、无血缘关系的献血个体作ABH多态性分析。结果 1)经唾液凝集抑制试验检测,结果显示在整个检测样本群中,A、B、O和AB血型比例分别为26.96%、29.57%、30.43%和13.04%,其中分泌型82.61%,非分泌型17.39%。2)在分泌型人群中,A、B、O和AB血型比例分别为28.42%、29.47%、28.42%和13.68%;在非分泌型人群中,A、B、O和AB血型比例分别为20%、30%、40%和10%。3)在同种血型人群中,男女间血型物质的效价差异不显著,无统计学意义;ABO血型中,血型A物质的效价高于B和H物质的效价,差异显著有统计学意义。4)经PCR-SSP及基因测序技术分析,结果显示在整个检测样本群中,FUT2的385位核苷酸类型为A/A、A/T和T/T的分布比例分别为41.74%、33.91%和24.35%。5)在分泌型人群中,FUT2的385位核苷酸类型为A/A、A/T和T/T的分布比例分别为50.53%、41.05%和8.42%;在非分泌型人群中,FUT2的385位核苷酸类型均为T/T;在非分泌型的个体中,检测出447G/447A突变。6)采用直接计数法计算基因频率并进行HardyWeinberg吻合度检验,显示本组样本A385T和G447A基因型分布符合Hardy-Weinberg平衡。结论 1)在ABO血型系统人群中,多数为分泌型者,少数非分泌型者。2)在同种血型人群中,男女间血型物质的效价差异不显著;血型A物质的效价高于B和O物质的效价,差异显著。3)基因FUT2的385A/A及385A/T在血型物质分泌时起决定作用;样本人群中G447A突变对血型物质分泌没有影响。
Objective To investigate the characteristics of ABH-secreting and non-secreting FUT2 gene polymorphism in ABL-positive and non-secretory ABH-rich and non-secretory blood of 115 unrelated Han donors in Zhoukou area, The feasibility of using PCR-SSP technology in the identification of ABO blood group. Methods A total of 115 saliva and peripheral blood samples from random blood donors were collected. Saliva sample agglutination inhibition test detection of secreted and non-secreted; using extraction kit genomic DNA, design A385T, and G447A two pairs of primers, polymerase chain reaction - sequence specific primers (PCR-SSP) technique and sequencing of 115 cases of healthy, unrelated blood donors for ABH polymorphism. Results 1) The test of salivary agglutination inhibition test showed that the proportion of A, B, O and AB blood group were 26.96%, 29.57%, 30.43% and 13.04% respectively, of which 82.61% were secretory Type 17.39%. 2) The proportion of A, B, O and AB blood group was 28.42%, 29.47%, 28.42% and 13.68% respectively in the secretory population. The proportion of A, B, O and AB in the non-secretory group was 20 %, 30%, 40% and 10%. 3) There was no significant difference in the titer of blood group substances between men and women in the same blood group, and the titer of blood group A was higher than that of B and H in ABO blood group, the difference was statistically significant significance. 4) The results of PCR-SSP and sequencing showed that the 385 nucleotide type of FUT2 was A / A, and the distribution ratios of A / T and T / T were 41.74% and 33.91 % And 24.35%. 5) In the secretory population, the 385 nucleotide type of FUT2 was A / A, and the distribution ratios of A / T and T / T were 50.53%, 41.05% and 8.42% respectively. In non-secretory population, FUT2 Of the 385 nucleotide types are T / T; in non-secreting individuals, a 447G / 447A mutation was detected. 6) Using direct counting method to calculate the frequency of genes and HardyWeinberg test, the distribution of A385T and G447A genotypes in this sample accord with Hardy-Weinberg equilibrium. Conclusion 1) In the ABO blood group system, the majority of people with secretions, a few non-secreting people. 2) In the same type of blood group, the titer of blood group substance between men and women is not significant; the titer of blood group A substance is higher than the titer of B and O substance, the difference is significant. 3) Gene FUT2 385A / A and 385A / T play a decisive role in the secretion of blood group substances; G447A mutation in the sample population has no effect on the secretion of blood group substances.