A multiplex PCR amplicon sequencing assay to screen genetic hearing loss variants in newborns

BMC Medical Genomics

Haiyan Yang¹  Jiale Xiang²  Hongyu Luo³  Zhiyu Peng³  Guiwei Zhang³  Junqing Zhang⁴ 

[1] BGI College, Zhengzhou University, 450001, Zhengzhou, China;School of Life Sciences, Zhengzhou University, 450001, Zhengzhou, China;BGI Education Center, University of Chinese Academy of Sciences, BGI Park, No.21 Hongan 3rd Street, Yantian District, 518083, Shenzhen, China;BGI Education Center, University of Chinese Academy of Sciences, BGI Park, No.21 Hongan 3rd Street, Yantian District, 518083, Shenzhen, China;BGI Genomics, BGI-Shenzhen, 518083, Shenzhen, China;BGI Genomics, BGI-Shenzhen, 518083, Shenzhen, China;Tianjin Medical Laboratory, BGI-Tianjin, BGI-Shenzhen, 300308, Tianjin, China;

关键词: Hearing loss;    Genetic screening;    GJB2;    SLC26A4;    MT-RNR1;   

DOI  :  10.1186/s12920-021-00906-1

来源: Springer

【 摘 要 】

BackgroundCongenital hearing loss is one of the most common birth defects. Early identification and management play a crucial role in improving patients’ communication and language acquisition. Previous studies demonstrated that genetic screening complements newborn hearing screening in clinical settings.MethodsWe developed a multiplex PCR amplicon sequencing assay to sequence the full coding region of the GJB2 gene, the most pathogenic variants of the SLC26A4 gene, and hotspot variants in the MT-RNR1 gene. The sensitivity, specificity, and reliability were validated via samples with known genotypes. Finally, a pilot study was performed on 300 anonymous dried blood samples.ResultsOf 103 samples with known genotypes, the multiplex PCR amplicon sequencing assay accurately identified all the variants, demonstrating a 100% sensitivity and specificity. The consistency is high in the analysis of the test–retest reliability and internal consistency reliability. In the pilot study, 12.3% (37/300) of the newborns were found to carry at least one pathogenic variant, including 24, 10, and 3 from the GJB2, SLC26A4, and MT-RNR1 gene, respectively. With an allele frequency of 2.2%, the NM_004004.6(GJB2):c.109G>A was the most prevalent variant in the study population.ConclusionThe multiplex PCR amplicon sequencing assay is an accurate and reliable test to detect hearing loss variants in the GJB2, SLC26A4, and MT-RNR1 genes. It can be used to screen genetic hearing loss in newborns.

【 授权许可】

CC BY