National Key Laboratory of Agricultural Microbiology, Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100086, China.
Curr Pharm Des. 2024;30(3):169-179. doi: 10.2174/0113816128276560231218090436.
Digital PCR (dPCR) is the latest technique that has become commercially accessible for various types of research. This method uses Taq polymerase in a standard polymerase chain reaction (PCR) to amplify a target DNA fragment from a complex sample, like quantitative PCR (qPCR) and droplet digital PCR (dd- PCR). ddPCR may facilitate microRNA (miRNA) measurement, particularly in liquid biopsy, because it has been proven to be more effective and sensitive, and in this method, ddPCR can provide an unprecedented chance for deoxyribonucleic acid (DNA) methylation research because of its capability to increase sensitivity and precision over conventional PCR-based methods. qPCR has also been found to be a valuable standard technique to measure both copy DNA (cDNA) and genomic DNA (gDNA) levels, although the finding data can be significantly variable and non-reproducible without relevant validation and verification of both primers and samples. The SYBR green quantitative real-time PCR (qPCR) method has been reported as an appropriate technique for quantitative detection and species discrimination, and has been applied profitably in different experiments to determine, quantify, and discriminate species. Although both TaqMan qRT-PCR and SYBR green qRT-PCR are sensitive and rapid, the SYBR green qRT-PCR assay is easy and the TaqMan qRT-PCR assay is specific but expensive due to the probe required. This review aimed to introduce dPCR, qPCR, SYBR green PCR kit, and digital PCR, compare them, and also introduce their advantages in the detection of different diseases.
数字 PCR(dPCR)是一种最新的技术,已经商业化应用于各种类型的研究。该方法在标准聚合酶链反应(PCR)中使用 Taq 聚合酶来扩增复杂样本中的目标 DNA 片段,如定量 PCR(qPCR)和液滴数字 PCR(dd-PCR)。dd-PCR 可能有助于 microRNA(miRNA)的测量,特别是在液体活检中,因为它已被证明更有效和敏感,并且在这种方法中,dd-PCR 可以通过增加灵敏度和精度为脱氧核糖核酸(DNA)甲基化研究提供前所未有的机会,因为它能够提高传统基于 PCR 的方法的灵敏度和精度。qPCR 也被发现是一种有价值的标准技术,可以测量 cDNA 和 gDNA 水平,尽管如果没有对引物和样本进行相关验证,发现的数据可能会有很大的差异和不可重复性。SYBR 绿色实时定量 PCR(qPCR)方法已被报道为一种用于定量检测和物种区分的合适技术,并已成功应用于不同实验中,以确定、定量和区分物种。尽管 TaqMan qRT-PCR 和 SYBR 绿色 qRT-PCR 都很敏感和快速,但 SYBR 绿色 qRT-PCR 检测法简单,而 TaqMan qRT-PCR 检测法则具有特异性但昂贵,因为需要探针。本综述旨在介绍 dPCR、qPCR、SYBR 绿色 PCR 试剂盒和数字 PCR,比较它们,并介绍它们在不同疾病检测中的优势。