Volume 23 Issue 4
Apr.  2025
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Article Navigation >  Chinese Journal of General Practice  > 2025  >  23(4): 623-626
PAN Shuqing, PAN Jiewen, BAO Youwei, PAN Xiaoli, ZHUANG Danyan. Application of the real-time fluorescence PCR melting curve method in gene screening for newborn hearing loss[J]. Chinese Journal of General Practice, 2025, 23(4): 623-626. doi: 10.16766/j.cnki.issn.1674-4152.003965
Citation: PAN Shuqing, PAN Jiewen, BAO Youwei, PAN Xiaoli, ZHUANG Danyan. Application of the real-time fluorescence PCR melting curve method in gene screening for newborn hearing loss[J]. Chinese Journal of General Practice, 2025, 23(4): 623-626. doi: 10.16766/j.cnki.issn.1674-4152.003965
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Application of the real-time fluorescence PCR melting curve method in gene screening for newborn hearing loss

doi: 10.16766/j.cnki.issn.1674-4152.003965

  • The Central Laboratory of Birth Defects Prevention and Control, Women and Children' s Hospital Affiliated to Ningbo University, Ningbo Key Laboratory for the Prevention and Treatment of Embryogenic Diseases, Ningbo Key Laboratory of Genomic Medicine and Birth Defects Prevention, Ningbo, Zhejiang 315012, China

Funds:

 2023KY1121

 2022-F26

  • Received Date: 2024-06-11
    Available Online: 2025-06-30
    Abstract
  •   Objective  To analyze the accuracy of the fluorescence PCR melting curve method in screening for newborn deafness genes, and to understand the variation types and frequencies of common deafness genes in the local population. It provides reference for gene screening and prevention of deafness in the future.  Methods  A retrospective analysis was conducted on 5 563 newborns and screened for deafness genes at Ningbo Women and Children' s Hospital Affiliated to Ningbo University, from November 2022 to October 2023. Firstly, the fluorescent PCR melting curve method was used to detect 15 mutation sites in four deafness genes (GJB2, SLC26A4, mitochondrial 12S rRNA, and GJB3), followed by Sanger sequencing to verify the positive samples.  Results  Among the 5 563 newborns, 256 cases of deafness gene mutation were detected, yielding a total detection rate of 4.60%. Mutations in the GJB2 gene were found in 156 cases (2.80%), in the SLC26A4 gene in 73 cases (1.31%), in the mitochondrial 12S rRNA gene in 10 cases (0.18%), and in the GJB3 gene in 16 cases (0.29%). Additionally, one case of complex heterozygous mutations was detected (0.02%). The positive samples were verified by Sanger sequencing, revealing 9 cases with inconsistent results. Therefore, the accuracy of the fluorescence PCR melting curve method is 96.48%.  Conclusion  The GJB2 gene and SLC26A4 gene are the main mutations associated with deafness in newborns in this region. Fluorescence PCR fusion curve method combined with Sanger sequencing method accurately detects genetic mutations related to deafness. This mothed is helpful for early detection of congenital, drug-induced, and delayed-onset deafness, playing an important role in the early prevention and treatment of deafness.

     

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