Laboratory of Human and Medical Genetics, Institute for Biological Sciences, Federal University of Pará, Belém, Pará, Brazil.
Am J Hum Biol. 2010 Sep-Oct;22(5):573-7. doi: 10.1002/ajhb.21047.
To assess alpha+-thalassemia deletion alleles, beta-thalassemia mutations and haplotypes linked to the HBBS cluster in a sample of 130 unrelated sickle cell anemia (SCA) patients (55% female) from Belém, Pará State, for their possible effects on the patients' survival. -alpha(3.7), -alpha(42), -alpha(20.5), and -(MED) alpha+-thalassemia deletion alleles were investigated using multiplex gap-PCR method. Characterization of beta-thalassemia mutations was made by direct genomic sequencing of the beta-globin gene amplified through polymerase chain reaction (PCR). Haplotypes were determined by analysis of six polymorphic restriction sites [(1) XmnI-5'gammaG, (2) HindIII-gammaG, (3) HindIII-gammaA, (4) HincII-psibeta, (5) HincII-3'psibeta, and (6) HinfI-5'beta] followed by restriction digestion and agarose gel electrophoresis. Twenty-one patients (16%) presented -alpha3.7 thalassemia. Sixteen of those (76%) were heterozygous (-alpha3.7/alphaalpha) and 5 (24%) were homozygous (-alpha3.7/-alpha3.7). -Alpha(4.2), -alpha(20.5) and -(MED) deletions were not found. Nine cases of sickle cell-beta thalassemia were found and four different beta-thal mutations were identified: beta(+) -88 (C>T), 3.8%; beta(+) codon 24 (T > A), 1.5%; beta(+) IVSI-110 (G > A), 0.7% and beta (IVSI-1 (G > A), 0.7%. No differences according to age were observed in -alpha(3.7) deletion, beta-thalassemia and HHBS haplotypes distribution. Our results suggest that although alpha- and beta-thalassemia and betaS haplotypes may have modulating effect on clinical expression and hematological parameters of SCA, these genetic variables probably have little influence on the subjects' survival.
为了评估 130 例无关联镰状细胞贫血（SCA）患者（55%为女性）样本中的 alpha+-珠蛋白生成障碍性贫血缺失等位基因、beta-地中海贫血突变和与 HBBS 簇相关的单倍型，以了解它们对患者生存的可能影响。使用多重间隙-PCR 方法检测 -alpha(3.7)、-alpha(42)、-alpha(20.5) 和 -(MED) alpha+-珠蛋白生成障碍性贫血缺失等位基因。通过聚合酶链反应（PCR）扩增的 beta-珠蛋白基因的直接基因组测序来确定 beta-地中海贫血突变的特征。通过分析六个多态性限制性位点 [(1) XmnI-5'gammaG，(2) HindIII-gammaG，(3) HindIII-gammaA，(4) HincII-psibeta，(5) HincII-3'psibeta 和 (6) HinfI-5'beta] 来确定单倍型，随后进行限制性消化和琼脂糖凝胶电泳。21 例患者（16%）存在 -alpha3.7 地中海贫血。其中 16 例（76%）为杂合子（-alpha3.7/alphaalpha），5 例（24%）为纯合子（-alpha3.7/-alpha3.7）。未发现 -alpha(4.2)、-alpha(20.5) 和 -(MED) 缺失。发现了 9 例镰状细胞-beta 地中海贫血病例，并鉴定出 4 种不同的 beta-地中海贫血突变：beta(+) -88（C>T），1.5%；beta(+) 密码子 24（T > A），0.7%；beta(+) IVSI-110（G > A），0.7%和 beta（IVSI-1（G > A），0.7%。-alpha(3.7)缺失、beta-地中海贫血和 HHBS 单倍型分布与年龄无关。我们的结果表明，尽管 alpha-和 beta-地中海贫血和 betaS 单倍型可能对 SCA 的临床表达和血液学参数具有调节作用，但这些遗传变量可能对受试者的生存影响不大。
