Zhang Zhang, Li Lian, Yao Juan
Department of Neurosurgery, Neurology Center, The First Affiliated Hospital of Hainan Medical University Haikou Hainan 571199 PR China
Nanobiosensing and Microfluidic Point-of-Care Testing, Key Laboratory of Luzhou, Department of Clinical Laboratory, The Affiliated Traditional Chinese Medicine Hospital, Southwest Medical University Luzhou Sichuan 646000 PR China
RSC Adv. 2025 Jan 2;15(1):75-82. doi: 10.1039/d4ra07286c.
Accurate, rapid, and multiplex SNP analysis holds significant clinical value. However, the inevitable nucleic acid extraction, involving centrifugation, heating, and magnetic separation, is often time-consuming. In this study, direct blood PCR was combined with dual-labelled probe-mediated melting curves to identify SNPs corresponding to MTHFR (C677T, rs#1801133 and A1298C, rs#1801131) and MTRR (A66G, rs#1801394) in a single tube. Our results indicated that nucleic acid extraction does not enhance gDNA concentrations. The results of the gDNA and whole blood samples were perfectly aligned with each other. The maximum volume tolerance for whole blood is 10%. Our method demonstrated robustness to hemolysis and temperature variations. The turnaround time of the test is <100 min with great potential value in clinical settings. Furthermore, this approach eliminates the need for complex preprocessing steps, simplifies the workflow and reduces potential sources of error. The efficiency and accuracy of this method make it a promising tool for routine clinical diagnostics and personalized medical applications. Additionally, the method's reliability and speed are crucial for effective patient management. The ability to obtain results quickly and accurately supports timely treatment decisions, which can be critical in many clinical scenarios. As a result, this technique has the potential to be widely adopted in clinical laboratories due to its practicality and effectiveness.
准确、快速且多重的单核苷酸多态性（SNP）分析具有重要的临床价值。然而，不可避免的核酸提取过程，包括离心、加热和磁分离，通常耗时较长。在本研究中，将直接血样聚合酶链反应（PCR）与双标记探针介导的熔解曲线相结合，在单个试管中鉴定与亚甲基四氢叶酸还原酶（MTHFR，C677T，rs#1801133和A1298C，rs#1801131）以及甲硫氨酸合成酶还原酶（MTRR，A66G，rs#1801394）相对应的SNP。我们的结果表明，核酸提取并未提高基因组DNA（gDNA）浓度。gDNA样本和全血样本的结果完全一致。全血的最大体积耐受量为10%。我们的方法显示出对溶血和温度变化具有稳健性。该检测的周转时间小于100分钟，在临床环境中具有巨大的潜在价值。此外，这种方法无需复杂的预处理步骤，简化了工作流程并减少了潜在的误差来源。该方法的效率和准确性使其成为常规临床诊断和个性化医疗应用的有前景的工具。此外，该方法的可靠性和速度对于有效的患者管理至关重要。快速准确地获得结果的能力支持及时的治疗决策，这在许多临床场景中可能至关重要。因此，由于其实用性和有效性，该技术有可能在临床实验室中被广泛采用。
