Background Congenital 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. Methods We 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. Results Of 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. Conclusion The 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.

The AT-hook is a small motif in DNA-binding protein that was first described in the high-mobility-group non-histone chromosomal protein HMG-I/Y. AT-hook family proteins play an important role in chromatin structure assembly, specific binding to target cells, transcription regulation and development regulation in other organisms. We identified 45 AT-hook genes from the rice genome by bioinformatics, analyzed the gene structure and chromosomal distribution, and examined the phylogenetic relationships among rice AT-hook proteins, finally constructing an unrooted tree from alignment of their full-length protein sequences in rice and Arabidopsis. AT-hook proteins grouped into 5 distinct clades (A-E), with no differences in structure or characteristics. The family members were recruited by chromosome replication. Digital gene expression analysis revealed that AT-hook genes mainly express in young panicles. We verified some of the gene expression by qRT-PCR.