INTRODUCTION: To evaluate a next-generation sequencing (NGS) workflow in the screening and diagnosis of thalassemia. METHODS: In this prospective study, blood samples were obtained from people undergoing genetic screening for thalassemia at our centre in Guangzhou, China. Genomic DNA was polymerase chain reaction (PCR)-amplified and sequenced using the Ion Torrent system and results compared with traditional genetic analyses. RESULTS: Of the 359 subjects, 148 (41%) were confirmed to have thalassemia. Variant detection identified 35 different types including the most common. Identification of the mutational sites by NGS were consistent with those identified by Sanger sequencing and Gap-PCR. The sensitivity and specificities of the Ion Torrent NGS were 100%. In a separate test of 16 samples, results were consistent when repeated ten times. CONCLUSION: Our NGS workflow based on the Ion Torrent sequencer was successful in the detection of large deletions and non-deletional defects in thalassemia with high accuracy and repeatability.
INTRODUCTION: To evaluate a next-generation sequencing (NGS) workflow in the screening and diagnosis of thalassemia. METHODS: In this prospective study, blood samples were obtained from people undergoing genetic screening for thalassemia at our centre in Guangzhou, China. Genomic DNA was polymerase chain reaction (PCR)-amplified and sequenced using the Ion Torrent system and results compared with traditional genetic analyses. RESULTS: Of the 359 subjects, 148 (41%) were confirmed to have thalassemia. Variant detection identified 35 different types including the most common. Identification of the mutational sites by NGS were consistent with those identified by Sanger sequencing and Gap-PCR. The sensitivity and specificities of the Ion Torrent NGS were 100%. In a separate test of 16 samples, results were consistent when repeated ten times. CONCLUSION: Our NGS workflow based on the Ion Torrent sequencer was successful in the detection of large deletions and non-deletional defects in thalassemia with high accuracy and repeatability.
Entities:
Keywords:
Ion Torrent; Next-generation sequencing; Sequencing; Thalassemia
Authors: Byung-Ok Choi; Soo Kyung Koo; Mi-Hyun Park; Hwanseok Rhee; Song-Ju Yang; Kyoung-Gyu Choi; Sung-Chul Jung; Han Su Kim; Young Se Hyun; Khriezhanuo Nakhro; Hye Jin Lee; Hae-Mi Woo; Ki Wha Chung Journal: Hum Mutat Date: 2012-07-05 Impact factor: 4.878