Nucleic Acid Tests for Clinical Translation

Abstract

Nucleic acids, including deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), are natural biopolymers composed of nucleotides that store, transmit, and express genetic information. Overexpressed or underexpressed as well as mutated nucleic acids have been implicated in many diseases. Therefore, nucleic acid tests (NATs) are extremely important. Inspired by intracellular DNA replication and RNA transcription, in vitro NATs have been extensively developed to improve the detection specificity, sensitivity, and simplicity. The principles of NATs can be in general classified into three categories: nucleic acid hybridization, thermal-cycle or isothermal amplification, and signal amplification. Driven by pressing needs in clinical diagnosis and prevention of infectious diseases, NATs have evolved to be a rapidly advancing field. During the past ten years, an explosive increase of research interest in both basic research and clinical translation has been witnessed. In this review, we aim to provide comprehensive coverage of the progress to analyze nucleic acids, use nucleic acids as recognition probes, construct detection devices based on nucleic acids, and utilize nucleic acids in clinical diagnosis and other important fields. We also discuss the new frontiers in the field and the challenges to be addressed.

CRISPR/Cas12a-Enhanced Loop-Mediated Isothermal Amplification for the Visual Detection of Shigella flexneri

“…At present, CRISPR/Cas plays an important role not only in immunity but also in gene editing, gene therapy, and especially in IVD ( Kaminski et al, 2021 ). In IVD, a large number of culture-, biochemical-, serological-, and molecular-based (nucleic acid) diagnostic methods were established for the detection of pathogens, and molecular diagnosis has gradually replaced the traditional biochemical and serological detection and occupies a higher market share in the field of IVD ( Min Li et al, 2021 ). Nucleic acid amplification in molecular diagnostics can be divided into constant-temperature amplification (e.g., LAMP and RPA) and variable-temperature amplification (e.g., PCR and qPCR) according to the temperature.…”

“…Constant-temperature amplification allows nucleic acid amplification at a constant temperature, which is non-instrument dependent. CRISPR/Cas, especially Cas12, Cas13, and Cas14, were widely used in molecular diagnosis combined with nucleic acid amplification ( Min Li et al, 2021 ), and as a novel molecular tool, it has great potential in IVD applications.…”

Emerging biosensing and transducing techniques for potential applications in point-of-care diagnostics

“…The development of PCR and DNA sequencing methods have made it possible to diagnose many infectious and genetic diseases rapidly and accurately, and to monitor their therapeutic effects. 46 These methods, however, have trade-offs between costs, sensitivity, selectivity and efficiency. Therefore, there is an urgent need for strategies capable of performing highly sensitive and selective single-base detection of nucleic acids.…”

Single-Molecule Detection of Nucleic Acids via Liposome Signal Amplification in Mass Spectrometry

“…As one of the basic life molecules, nucleic acids play a pivotal role in the storage, transmission, and expression of genetic information. Nucleic acids are closely related to various clinical diseases and have become an important basis for their clinical diagnosis and treatment [ 1 , 2 ]. Currently, nucleic acid detection is performed via signal amplification methods.…”