Naja R P, Kaspar H, Shbaklo H, Chakar N, Makhoul N J, Zalloua P A
Genetics Research Laboratory, Chronic Care Center, Beirut, Lebanon.
Am J Hematol. 2004 Apr;75(4):220-4. doi: 10.1002/ajh.20013.
Beta-thalassemia is the most common genetic disorder in the Lebanese population. Of the 200 different mutations in the beta-globin gene that leads to thalassemia, the IVSI-110 (29.87%), IVSI-6 (20.74%), IVSI-1 (14.07%), IVSII-1 (9.13%), Cd29 (9.13%), and Cd30 (3.95%) mutations are the most frequent among Lebanese thalassemic patients. These mutations are also present at high frequencies in the East Mediterranean region. Due to this high prevalence of certain beta-thalassemia mutations, a rapid technique for the prenatal diagnosis of these mutations was implemented. The technique used is based on Real-Time PCR quantification and melting curve analysis of the amplified fragment using the LightCycler. The DNA samples used for amplification were obtained from CVS or amniotic fluid. Six mutations were easily and efficiently detected using only 3 sets of probes. With this method, mutant genotypes can be easily distinguished from normal alleles. In prenatal diagnosis, the accuracy and the speed of testing are paramount. The method of prenatal beta-thalassemia mutations detection described here is efficient and fast, with the entire procedure including DNA preparation taking less than half a workday. It is safe, does not involve radioactivity, and is accurate showing 100% concordance with conventional DNA sequencing methods.
β地中海贫血是黎巴嫩人群中最常见的遗传性疾病。在导致地中海贫血的β珠蛋白基因的200种不同突变中，IVSI-110（29.87%）、IVSI-6（20.74%）、IVSI-1（14.07%）、IVSII-1（9.13%）、Cd29（9.13%）和Cd30（3.95%）突变在黎巴嫩地中海贫血患者中最为常见。这些突变在东地中海地区也以高频率存在。由于某些β地中海贫血突变的高流行率，实施了一种快速的这些突变的产前诊断技术。所使用的技术基于实时PCR定量和使用LightCycler对扩增片段进行熔解曲线分析。用于扩增的DNA样本取自绒毛取样（CVS）或羊水。仅使用3组探针就能轻松且高效地检测出6种突变。通过这种方法，可以轻松地将突变基因型与正常等位基因区分开来。在产前诊断中，检测的准确性和速度至关重要。这里描述的产前β地中海贫血突变检测方法高效且快速，整个过程包括DNA制备在内耗时不到半个工作日。它安全，不涉及放射性，并且准确，与传统DNA测序方法的一致性达100%。
