Development and clinical application of a low-cost high-resolution melting curve assay for CD36 gene variant
ZhengTing Liu1, JiaLi Huang2, Jianxin Zhen3
1Department of Clinical Laboratory, Ganzhou People's Hospital, 16 Meiguan Avenue, Zhanggong District, Ganzhou, China.
Background:
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.
Materials And Methods:
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.
Results:
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).
Discussion:
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.
