Hassan S M, Vossen R H A M, Chessa R, den Dunnen J T, Bakker E, Giordano P C, Harteveld C L
Molecular Genetic Laboratory, National Genetic Centre, Muscat, Oman; Department of Human and Clinical Genetics, Leiden University Medical Centre, Leiden, The Netherlands.
Leiden Genome Technology Center (LGTC), Human and Clincal Genetics, Leiden University Medical Center, Leiden, The Netherlands.
Blood Cells Mol Dis. 2014 Sep;53(3):133-7. doi: 10.1016/j.bcmd.2014.05.002. Epub 2014 May 28.
Hemoglobinopathies, such as sickle cell disease (SCD) and beta-thalassemia major (TM), are severe diseases and the most common autosomal recessive condition worldwide and in particular in Oman. Early screening and diagnosis of carriers are the key for primary prevention. Once a country-wide population screening program is mandated by law, a sequencing technology that can rapidly confirm or identify disease-causing mutations for a large number of patients in a short period of time will be necessary. While Sanger sequencing is the standard protocol for molecular diagnosis, next generation sequencing starts to become available to reference laboratories. Using the Ion Torrent PGM sequencer, we have analyzed a cohort of 297 unrelated Omani cases and reliably identified mutations in the beta-globin (HBB) gene. Our model study has shown that Ion Torrent PGM can rapidly sequence such a small gene in a large number of samples using a barcoded uni-directional or bi-directional sequence methodology, reducing cost, workload and providing accurate diagnosis. Based on our results we believe that the Ion Torrent PGM sequencing platform, able to analyze hundreds of patients simultaneously for a single disease gene can be a valid molecular screening alternative to ABI sequencing in the diagnosis of hemoglobinopathies and other genetic disorders in the near future.
血红蛋白病,如镰状细胞病(SCD)和重型β地中海贫血(TM),是严重的疾病,也是全球尤其是阿曼最常见的常染色体隐性疾病。对携带者进行早期筛查和诊断是一级预防的关键。一旦依法强制实施全国性人群筛查计划,就需要一种能够在短时间内快速确认或识别大量患者致病突变的测序技术。虽然桑格测序是分子诊断的标准方案,但新一代测序技术已开始可供参考实验室使用。我们使用Ion Torrent PGM测序仪分析了297例不相关的阿曼病例,并可靠地鉴定出β珠蛋白(HBB)基因中的突变。我们的模型研究表明,Ion Torrent PGM可以使用条形码单向或双向序列方法在大量样本中快速对如此小的基因进行测序,降低成本、工作量并提供准确诊断。基于我们的结果,我们相信能够同时对数百名患者的单个疾病基因进行分析的Ion Torrent PGM测序平台在不久的将来可以成为血红蛋白病和其他遗传疾病诊断中替代ABI测序的有效分子筛查方法。