Liu ZhengTing, Huang JiaLi, Zhen Jianxin, Chen ShangLiang
Department of Clinical Laboratory, Ganzhou People's Hospital, 16 Meiguan Avenue, Zhanggong District, Ganzhou, China.
Department of Clinical Laboratory, the First Affiliated Hospital of Shantou University Medical College, 57 Changping Road, Jinping District, Shantou, China.
Mol Biol Rep. 2025 Aug 7;52(1):805. doi: 10.1007/s11033-025-10891-8.
CD36 gene variants contribute to immune thrombocytopenia and are commonly observed in Asian populations. Detection of CD36 gene variants in clinical laboratories is currently limited, necessitating the development of a rapid and cost-effective screening method. Here, we presented a high-resolution melting curve assay for CD36 gene variant detection and evaluated its clinical applicability.
We designed a high-resolution melting (HRM)-based method for screening for CD36 gene variants and assessed its specificity, sensitivity, and repeatability via recombinant plasmids. We subsequently employed the HRM assay to analyze 758 clinical samples, with some results validated by DNA sequencing.
The HRM assay enabled the complete detection of CD36 gene variants, even at allele frequencies as low as 15%, with a detection limit of 1 pg of CD36 gene DNA. Among the 758 patients tested, 21 (2.77%) exhibited CD36 gene variants, with the most prevalent variant being 320-330delAC. The rates of CD36 gene variant detection were significantly higher in hematological patients (3.08%, 9/292), pregnant women (3.70%, 3/81), and organ transplant patients (4.27%, 8/187) than in the healthy control group (0.51%, 1/198).
Our findings demonstrate that high-resolution melting analysis is a highly sensitive and specific method for identifying CD36 gene polymorphisms. This low-cost HRM assay is suitable for large-scale CD36 gene variant screening in clinical laboratories.
CD36基因变异与免疫性血小板减少症有关，在亚洲人群中较为常见。目前临床实验室对CD36基因变异的检测有限，因此需要开发一种快速且经济高效的筛查方法。在此，我们介绍了一种用于检测CD36基因变异的高分辨率熔解曲线分析方法，并评估了其临床适用性。
我们设计了一种基于高分辨率熔解（HRM）的方法来筛查CD36基因变异，并通过重组质粒评估其特异性、敏感性和重复性。随后，我们采用HRM分析方法对758份临床样本进行分析，部分结果通过DNA测序进行验证。
HRM分析能够完全检测到CD36基因变异，即使等位基因频率低至15%，CD36基因DNA的检测限为1 pg。在758例检测患者中，21例（2.77%）表现出CD36基因变异，最常见的变异是320-330delAC。血液系统疾病患者（3.08%，9/292）、孕妇（3.70%，3/81）和器官移植患者（4.27%，8/187）的CD36基因变异检出率显著高于健康对照组（0.51%，1/198）。
我们的研究结果表明，高分辨率熔解分析是一种用于识别CD36基因多态性的高度灵敏且特异的方法。这种低成本的HRM分析方法适用于临床实验室大规模筛查CD36基因变异。
