Department of Chemistry and Beckman Institute, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA.
Anal Chem. 2012 Feb 21;84(4):1975-80. doi: 10.1021/ac203014s. Epub 2012 Feb 6.
DNA detection is commonly used in molecular biology, pathogen analysis, genetic disorder diagnosis, and forensic tests. While traditional methods for DNA detection such as polymerase chain reaction (PCR) and DNA microarrays have been well developed, they require sophisticated equipment and operations, and thus it is still challenging to develop a portable and quantitative DNA detection method for the public use at home or in the field. Although many other techniques and devices have been reported to make the DNA detection simple and portable, very few of them are currently accessible to the public for quantitative DNA detection because of either the requirement of laboratory-based instrument or lack of quantitative detection. Herein we report application of personal glucose meters (PGMs), which are widely available, low cost, and simple to use, for quantitative detection of DNA, including a hepatitis B virus DNA fragment. The quantification is based on target-dependent binding of cDNA-invertase conjugate with the analyte DNA, thereby transforming the concentration of DNA in the sample into glucose through invertase-catalyzed hydrolysis of sucrose. Instead of amplifying DNA strands through PCR, which is vulnerable to contaminations commonly encountered for home and field usage, we demonstrate here signal amplifications based on enzymatic turnovers, making it possible to detect 40 pM DNA using PGM that can detect glucose only at the mM level. The method also shows excellent selectivity toward single nucleotide mismatches.
DNA 检测在分子生物学、病原体分析、遗传疾病诊断和法医检测中得到了广泛应用。尽管聚合酶链式反应(PCR)和 DNA 微阵列等传统的 DNA 检测方法已经得到了很好的发展,但它们需要复杂的设备和操作,因此,开发一种便携式和定量的 DNA 检测方法,供公众在家中或现场使用,仍然具有挑战性。虽然已经有许多其他技术和设备被报道用于简化和便携式 DNA 检测,但由于需要基于实验室的仪器或缺乏定量检测,目前很少有技术和设备可供公众进行定量 DNA 检测。在这里,我们报告了个人血糖仪(PGM)在定量检测 DNA 中的应用,包括乙型肝炎病毒 DNA 片段。这种定量检测是基于 cDNA-转化酶缀合物与分析物 DNA 的靶依赖性结合,从而通过转化酶催化蔗糖水解将样品中 DNA 的浓度转化为葡萄糖。与 PCR 等容易受到家庭和现场常见污染的 DNA 链扩增方法不同,我们在这里展示了基于酶循环的信号放大,使得可以使用只能检测到 mM 水平葡萄糖的 PGM 检测到 40 pM 的 DNA。该方法还表现出对单个核苷酸错配的优异选择性。