Genotyping of Multiple Clinical Samples with a Combined Direct PCR and Magnetic Lateral Flow Assay.
Genotyping of Multiple Clinical Samples with a Combined Direct PCR and Magnetic Lateral Flow Assay.
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结合直接 PCR 和磁力横向流动分析对多个临床样本进行基因分型
DOI：
10.1016/j.isci.2018.09.005
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发表时间：
2018-09-28
期刊：
影响因子：
5.8
通讯作者：
Cui Y
中科院分区：
文献类型：
--
作者：
Zhang C;Liu X;Yao Y;Liu K;Hui W;Zhu J;Dou Y;Hua K;Peng M;Wang Z;Vermorken AJM;Cui Y
开发一种灵敏、低成本和易于使用的用于基因分型的即时检测系统对于告知治疗决策和预测基础疾病的风险非常重要。传统方法通常需要复杂的操作程序以及昂贵和复杂的仪器。在这里，我们报告了一种通用方法，使我们能够直接检测多种样本类型的基因型，而无需DNA纯化。此外，PCR结果可以进一步定量分析的基础上的磁侧流分析（MLFA）系统，这避免了传统的核酸生物传感器所需的多个步骤。结果表明，该方法可以在小体积样品中同时鉴定出3种基因型，准确率为100%，灵敏度为1.0 × 102 cells/μL，优于金标准方法。我们认为，直接PCR-MLFA系统代表了便携式，敏感的生物医学平台的发展的重大进展。多个样本（血液、DBS、唾液和口腔拭子）直接用于PCR模板结合直接PCR和磁性侧向流分析用于ALDH 2基因分型基因型可在90分钟内获得临床遗传学;分析化学;流体学;生化分析
Developing a sensitive, low-cost, and easy-to-use point-of-care testing system for genotyping is important for informing treatment decisions and predicting the risk of underlying diseases. Conventional methods normally require complex operational procedures as well as expensive and sophisticated instruments. Here, we report a general approach that enables us to detect the genotype of multiple sample types directly without DNA purification. Moreover, the PCR results can be further quantitatively analyzed based on a magnetic lateral flow assay (MLFA) system, which avoids multiple steps needed for conventional nucleic acid biosensors. As a demonstration, we show that three genotypes of aldehyde dehydrogenase 2 (ALDH2) can be identified using a small volume of sample with an accuracy of 100% and a sensitivity of 1.0 × 102 cells/μL, which are better than those of the gold standard methods. We believe that the direct PCR-MLFA system represents a significant advance toward the development of portable, sensitive biomedical platforms. Multiple samples (blood, DBS, saliva and buccal swab) for PCR template directly Combined direct PCR and magnetic lateral flow assay for ALDH2 genotyping Genotypes can be obtained within 90 min Clinical Genetics; Analytical Chemistry; Fluidics; Biochemical Analysis
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