Villa A, Drago F, Mistò R, Morelati F, Poli F, Sirchia G
Centro Trasfusionale e di Immunologia dei Trapianti, IRCCS Ospedale Maggiore, Milano, Italy.
Haematologica. 1996 Nov-Dec;81(6):492-6.
Traditional ABO blood group serology is based on the immunoreactivity of antisera with the carbohydrate A, B and H antigens. Progress in the molecular biology of the ABO system has recognized the molecular basis of the red cell (RBC) antigens and has provided a genetic model for ABO polymorphism at the molecular level. Recently, this genetic model was tested in a large number of individuals.
In this study we applied DNA analysis to determine the frequency of ABO genotypes in a group of blood donors for whom the ABO type was known. Two hundred and fifty healthy Italian blood donors were analyzed using polymerase chain reaction (PCR) to amplify two different regions of genomic DNA, each of which contained a different nucleotide polymorphism. The amplified product was digested with 4 restriction enzymes that revealed differences among A, B and O individuals. To analyze the genes at polymorphic sites 261 and 703 we used the restriction enzymes BstE II and Kpn I, and Hpa II and Alu I and compared the PCR determined genotypes to serologically determined phenotypes.
The results were consistent for all unrelated individuals; however, 2 of 100 individuals with the 0 phenotype carried one allele that differed from the proposed genetic model. This novel O allele, termed 0(2) by Yamamoto et al., was found in our series with a frequency of 1%. The blood group AB0 genotype of 250 healthy Italian blood donors was: 13 AA/AO(2), 37 AO(1), 11 BB, 39 B0(1), 50 AB, 98 0(1)0(1) and 2 0(1)0(2). This method should be applicable not only in forensic medicine but also in immunohematology when serology fails.
传统的ABO血型血清学基于抗血清与碳水化合物A、B和H抗原的免疫反应性。ABO系统分子生物学的进展已经认识到红细胞（RBC）抗原的分子基础，并在分子水平上为ABO多态性提供了遗传模型。最近，这个遗传模型在大量个体中得到了验证。
在本研究中，我们应用DNA分析来确定一组已知ABO血型的献血者中ABO基因型的频率。使用聚合酶链反应（PCR）对250名健康的意大利献血者进行分析，以扩增基因组DNA的两个不同区域，每个区域都包含不同的核苷酸多态性。扩增产物用4种限制性内切酶消化，这些酶揭示了A、B和O个体之间的差异。为了分析多态性位点261和703处的基因，我们使用了限制性内切酶BstE II和Kpn I，以及Hpa II和Alu I，并将PCR确定的基因型与血清学确定的表型进行比较。
所有无关个体的结果均一致；然而，100名O型表型个体中有2人携带一个与所提出的遗传模型不同的等位基因。这个新的O等位基因，被山本等人称为O(2)，在我们的研究系列中的频率为1%。250名健康意大利献血者的ABO血型基因型为：13例AA/AO(2)、37例AO(1)、11例BB、39例B0(1)、50例AB、98例O(1)O(1)和2例O(1)O(2)。这种方法不仅应适用于法医学，而且在血清学无法进行时也适用于免疫血液学。
