Division of Molecular Diagnostics, Department of Laboratory Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.
Clin Chim Acta. 2012 Dec 24;414:197-201. doi: 10.1016/j.cca.2012.09.012. Epub 2012 Sep 18.
High-resolution melting (HRM) analysis is a feasible and powerful method for mutation scanning of sequence variants. Denatured doubled-stranded DNA can be detected in fluorescence changes by increasing the melting temperature and wild-type and heterozygous samples can be easily differentiated in the melting plots. HRM analysis represents the next generation of mutation-scanning technology and offers considerable time and cost savings compared to other screening methods. HRM analysis is a closed-tube method, indicating that polymerase chain reaction amplification and subsequent analysis are sequentially performed in the well, making HRM analysis more convenient than other scanning methodologies. Taken together, HRM analysis can be used for high-throughput mutation screening for research, as well as for molecular diagnostic and clinical purposes. This review summarizes the effectiveness of HRM analysis in the diagnosis of autosomal recessive, dominant, and X-linked genetic disorders. Notably, we will also discuss the limitations of HRM analysis and how to overcome them.
高分辨率熔解（HRM）分析是一种用于检测序列变异的可行且强大的方法。通过增加熔点，可以检测到变性的双链 DNA 荧光变化，并且可以在熔解图谱中轻松区分野生型和杂合样本。HRM 分析代表了下一代突变扫描技术，与其他筛选方法相比，可节省大量的时间和成本。HRM 分析是一种闭管方法，这意味着聚合酶链反应扩增和随后的分析在孔中依次进行，使 HRM 分析比其他扫描方法更方便。总之，HRM 分析可用于高通量突变筛选，无论是用于研究目的，还是用于分子诊断和临床目的。本综述总结了 HRM 分析在常染色体隐性、显性和 X 连锁遗传疾病诊断中的有效性。值得注意的是，我们还将讨论 HRM 分析的局限性以及如何克服这些局限性。
