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Mutation spectrum of COL1A1/COL1A2 screening by high-resolution melting analysis of Chinese patients with osteogenesis imperfecta

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Abstract

High-resolution melting (HRM) analysis has been shown to be a time-saving method for the screening of genetic variants. To increase the precision of the diagnosis of osteogenesis imperfecta (OI), we used HRM to explore COL1A1/COL1A2 mutations in 87 Chinese OI patients and to perform population-based studies of the relationships between their genotypes and phenotypes. Peripheral blood samples were collected from the 87 non-consanguineous probands. The coding regions and exon boundaries of COL1A1/COL1A2 were detected by HRM and confirmed by Sanger sequencing. The functional effects of mutations were predicted through bioinformatic tools. Mutations were detected in 70.3% of familial cases and 40% of sporadic cases (p < 0.01). Compared with COL1A1 mutations, patients with COL1A2 mutations were more prone to severe phenotypes. Helical mutations (caused by substitution of the glycine within the Gly–X–Y triplet domain) were more likely to occur in patients with type III and IV (p < 0.05). Haploinsufficiency mutations (caused by frameshift, nonsense, and splice-site mutations) appeared more frequently in patients with type I (p < 0.05). Compared with the Sanger sequencing and whole exome sequencing (WES), HRM was found to reduce total costs by 78%– 80% in patients who had a positive HRM separate melting curve. Our findings suggest that HRM would greatly benefit small and understaffed hospitals and laboratories, and would facilitate the accurate diagnosis and early treatment of OI in remote and less developed regions.

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Acknowledgments

The authors would like to thank Dr. Ren Xiuzhi and nurse Chen Mei for providing clinical diagnosis and control blood samples.

Funding

This study was supported by research grants from National Key R&D Program of China (2017YFC1001904); National Natural Science Foundation of China (21647008); Tianjin Science and Technology Support Program (16YFZCSY00900).

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Author notes

  1. Mingyan Ju, Xue Bai and Tianke Zhang have contributed equally to this work.

Authors and Affiliations

  1. Department of Genetics, Tianjin Medical University, Tianjin, 300070, People’s Republic of China

    Mingyan Ju, Keqiu Li & Guang Li

  2. Department of Medical Laboratory, Tianjin Hospital, Tianjin, 300000, People’s Republic of China

    Xue Bai, Xiaoli Chang, Shizhen Guan & Yi Wang

  3. Colorectal Surgery, Tianjin People’s Hospital, Tianjin, 300000, People’s Republic of China

    Tianke Zhang

  4. School of Basic Medical Sciences, Tianjin Medical University, Tianjin, 300070, People’s Republic of China

    Yunshou Lin, Li Yang & Huaiyu Zhou

  5. Orthopedic Ward III, Wuqing People’s Hospital, Tianjin, 300000, People’s Republic of China

    Xiuzhi Ren

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  1. Mingyan Ju
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Correspondence to Guang Li.

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Ju, M., Bai, X., Zhang, T. et al. Mutation spectrum of COL1A1/COL1A2 screening by high-resolution melting analysis of Chinese patients with osteogenesis imperfecta. J Bone Miner Metab 38, 188–197 (2020). https://doi.org/10.1007/s00774-019-01039-3

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