Li Zongzhe, Huang Jin, Zhao Jinzhao, Chen Chen, Wang Hong, Ding Hu, Wang Dao Wu, Wang Dao Wen
Departments of Internal Medicine and Gene Therapy Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1095# Jiefang Ave, Wuhan 430030, China.
J Transl Med. 2014 Jun 17;12:173. doi: 10.1186/1479-5876-12-173.
Rapidly determining the complex genetic basis of Hypertrophic cardiomyopathy (HCM) is vital to better understanding and optimally managing this common polygenetic cardiovascular disease.
A rapid custom Ion-amplicon-resequencing assay, covering 30 commonly affected genes of HCM, was developed and validated in 120 unrelated patients with HCM to facilitate genetic diagnosis of this disease. With this HCM-specific panel and only 20 ng of input genomic DNA, physicians can, for the first time, go from blood samples to variants within a single day.
On average, this approach gained 595628 mapped reads per sample, 95.51% reads on target (64.06 kb), 490-fold base coverage depth and 93.24% uniformity of base coverage in CDS regions of the 30 HCM genes. After validation, we detected underlying pathogenic variants in 87% (104 of 120) samples. Tested seven randomly selected HCM genes in eight samples by Sanger sequencing, the sensitivity and false-positive-rate of this HCM panel was 100% and 5%, respectively.
This Ion amplicon HCM resequencing assay provides a currently most rapid, comprehensive, cost-effective and reliable measure for genetic diagnosis of HCM in routinely obtained samples.
快速确定肥厚型心肌病（HCM）复杂的遗传基础对于更好地理解和优化管理这种常见的多基因心血管疾病至关重要。
开发了一种快速定制的离子扩增子重测序检测方法，涵盖30个HCM常见受累基因，并在120例无亲缘关系的HCM患者中进行了验证，以促进该疾病的基因诊断。使用这种HCM特异性检测板，仅需20 ng输入基因组DNA，医生首次能够在一天内从血样获得变异信息。
平均而言，该方法每个样本获得595628条比对读数，95.51%的读数在目标区域（64.06 kb），30个HCM基因的编码区碱基覆盖深度为490倍，碱基覆盖均匀性为93.24%。验证后，我们在87%（120例中的104例）样本中检测到潜在的致病变异。通过Sanger测序对8个样本中的7个随机选择的HCM基因进行检测，该HCM检测板的灵敏度和假阳性率分别为100%和5%。
这种离子扩增子HCM重测序检测方法为在常规获取的样本中进行HCM基因诊断提供了目前最快速、全面、经济高效且可靠的手段。
