College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, 310058, China.
Key Laboratory of Microbiol Technology and Bioinformatics of Zhejiang Province, Zhejiang Institute of Microbiology, Hangzhou, 310012, China.
Biosens Bioelectron. 2020 Apr 1;153:112049. doi: 10.1016/j.bios.2020.112049. Epub 2020 Jan 27.
Efficient and rapid detection of pathogens plays an important role in food safety, disease prevention, diagnosis and environmental monitoring. The traditional method for pathogen detection is plate culturing, consuming lots of time on separating, culturing and identifying pathogens by morphological characteristics, biochemical and serological reactions. It is a great advantage to take nucleic acids of pathogens as targets for detection because of higher specificity. The polymerase chain reaction (PCR) greatly shortens the time of pathogen detection but it is heavily dependent on temperature control instruments. Although isothermal amplification overcomes the defects of temperature control, it requires multiple enzymes or complex primers. Here, we summarize the recent advances in the amplification free methods for pathogen detection which are well developed for their simplicity, sensitivity and rapidity. Without nucleic acid amplification, we can directly detect the original nucleic acids of the samples rather than amplified nucleic acids. The amplification free methods for nucleic acid detection are mainly classified into electrochemical biosensors, optical biosensors and piezoelectric plate biosensors. This article describes the principles and compares the advantages and disadvantages of these methods. We further discuss the challenges and directions of this field, providing an overview for future researchers.
病原体的高效快速检测在食品安全、疾病预防、诊断和环境监测中起着重要作用。传统的病原体检测方法是平板培养,通过形态特征、生化和血清学反应来分离、培养和鉴定病原体需要大量的时间。以病原体的核酸为检测目标具有更高的特异性,这是一个很大的优势。聚合酶链反应(PCR)极大地缩短了病原体检测的时间,但严重依赖于温度控制仪器。虽然等温扩增克服了温度控制的缺陷,但它需要多种酶或复杂的引物。在这里,我们总结了近年来病原体检测中无扩增方法的最新进展,这些方法因其简单、灵敏和快速而得到了很好的发展。无需核酸扩增,我们可以直接检测样品中原核酸,而不是扩增的核酸。核酸检测的无扩增方法主要分为电化学生物传感器、光学生物传感器和压电板生物传感器。本文描述了这些方法的原理,并比较了它们的优缺点。我们进一步讨论了该领域的挑战和方向,为未来的研究人员提供了一个概述。