Puehringer Helene, Najmabadi Hossein, Law Hai-Yang, Krugluger Walter, Viprakasit Vip, Pissard Serge, Baysal Erol, Taher Ali, Farra Chantal, Al-Ali Amein, Al-Ateeq Suad, Oberkanins Christian
ViennaLab Diagnostics GmbH, Vienna, Austria.
Clin Chem Lab Med. 2007;45(5):605-10. doi: 10.1515/CCLM.2007.125.
alpha-Thalassemia is a worldwide disease and considered to be a major public health problem in countries within the so-called thalassemia belt. The complex genetics of alpha-thalassemias requires diagnostic methods with the capacity to screen rapidly and accurately for common causative mutations.
We developed and validated a reverse-hybridization assay (Alpha-Globin StripAssay) for the rapid and simultaneous detection of 21 alpha-globin mutations: two single gene deletions (-alpha(3.7); -alpha(4.2)), five double gene deletions [--(MED); --(SEA); --(THAI); --(FIL); -(alpha)(20.5)], alpha alpha alpha(anti-3.7) gene triplication, two point mutations in the alpha1 gene (cd 14 G>A; Hb Adana) and 11 point mutations in the alpha2 gene (initiation cd T>C; cd 19 -G; IVS1 -5nt; cd 59 G>A; Hb Quong Sze; Hb Constant Spring; Hb Icaria; Hb Pakse; Hb Koya Dora; polyA-1; polyA-2).
Reliable genotyping of recombinant mutant clones and reference DNA samples was achieved by means of two corresponding test strips presenting parallel arrays of allele-specific oligonucleotides. The entire procedure from blood sampling to the identification of mutations required less than 6 h, and hybridization/detection was manual or automated. The diagnostic potential of this Alpha-Globin StripAssay was carefully evaluated on 272 pre-typed samples in a multicenter validation study. In 96.14% of the cases, StripAssay typing was completely concordant with the reference methods.
The Alpha-Globin StripAssay proved to be a fast, easy-to-perform and reliable screening method to identify >90% of alpha-globin mutations in endemic areas worldwide.
α地中海贫血是一种全球性疾病，在所谓的地中海贫血带国家被视为一个主要的公共卫生问题。α地中海贫血复杂的遗传学特性需要具备快速、准确筛查常见致病突变能力的诊断方法。
我们开发并验证了一种反向杂交检测法（α珠蛋白条带检测法），用于快速同时检测21种α珠蛋白突变：两种单基因缺失（-α(3.7)；-α(4.2)）、五种双基因缺失[--(MED)；--(SEA)；--(THAI)；--(FIL)；-(α)(20.5)]、ααα(抗-3.7)基因三倍体、α1基因中的两种点突变（第14密码子G>A；Hb阿德纳）以及α2基因中的11种点突变（起始密码子T>C；第19密码子-G；内含子1 -5nt；第59密码子G>A；Hb泉寺；Hb Constant Spring；Hb伊卡里亚；Hb巴色；Hb小田原；多聚腺苷酸-1；多聚腺苷酸-2）。
通过两条呈现等位基因特异性寡核苷酸平行阵列的相应检测条带，实现了重组突变克隆和参考DNA样本的可靠基因分型。从采血到突变鉴定的整个过程耗时不到6小时，杂交/检测可手动或自动进行。在一项多中心验证研究中，对272个预先分型的样本仔细评估了这种α珠蛋白条带检测法的诊断潜力。在96.14%的病例中，条带检测法分型与参考方法完全一致。
α珠蛋白条带检测法被证明是一种快速、易于操作且可靠的筛查方法，可用于识别全球流行地区90%以上的α珠蛋白突变。
