XÁC ĐỊNH KIỂU GEN THALASSEMIA Ở BỆNH NHÂN CÓ CÔNG THỨC MÁU NGHI NGỜ BẰNG REAL-TIME PCR ĐA KÊNH PHÂN TÍCH ĐƯỜNG CONG NÓNG CHẢY

Abstract

Objectives: To determine the carrier rate, mutation spectrum (19 variants), and hematological characteristics in patients with suspected thalassemia based on abnormal complete blood count (CBC) in Northern Vietnam, using multichannel melting curve real-time PCR. Methods: Cross-sectional descriptive study on 116 patients (01/2023–06/2025) at Chemedic Laboratory. Genomic DNA was extracted from peripheral blood and genotyped using multichannel fluorescent real-time PCR with melting curve analysis (Thalassemia Gene Assay kit, Yaneng Bioscience, IVD-certified) on CFX96 Touch/DTprime-5 systems. Data analyzed with OriginPro 2024; Mann-Whitney U test for hematological comparisons. Results: Carrier rate 50.9% (59/116); α-thalassemia 29.3%, β-thalassemia 18.9%, α+β co-inheritance 2.6%. Total 66 mutation events: --SEA 37.9%, HbE 18.2%, CD41/42 12.1%. Carriers exhibited higher RBC (5.41 ± 0.55 vs 4.87 ± 1.01 ×10¹²/L; p=0.006), lower MCHC (295 ± 19.9 vs 306 ± 21.7 g/L; p=0.003), and higher RDW-CV (17.4 ± 3.2 vs 16.6 ± 4.1%; p=0.056). Conclusion: High thalassemia carrier prevalence with --SEA and HbE predominance aligns with Southeast Asian epidemiology. Multichannel melting curve real-time PCR demonstrates high sensitivity (>99%) for simultaneous detection of 24 mutations, facilitating community screening and premarital genetic counseling.