Advances in Research on Isothermal Signal Amplification Mediated MicroRNA Detection of Clinical Samples: Application to Disease Diagnosis

Abstract

With the rapid development of modern molecular biology, microRNA (miRNA) has been demonstrated to be closely associated with the occurrence and development of tumors and holds significant promise as a biomarker for the early detection, diagnosis, and treatment of cancer and other diseases. Therefore, detecting miRNA and analyzing it to determine its biological functions are of great significance for the screening and diagnosis of diseases. However, the intrinsic characteristics of miRNAs, including their low abundance, short sequence lengths, and high family-specific sequence homology, render traditional detection methods such as Northern blot hybridization, microarray use, and reverse transcription quantitative PCR (RT-qPCR) inadequate for meeting the stringent requirements of clinical detection in biological samples, a task requiring accuracy, rapidity, high detection power, specificity, and cost-effectiveness. In recent years, a substantial amount of effort has been put into developing innovative methodologies to address these challenges. In this review, we aim to provide a comprehensive overview of the recent advancements in these methodologies and their applications in clinical biological sample detection for disease diagnosis.

Keywords: disease diagnosis; isothermal signal amplification; miRNA detection; point-of-care testing.

Figure 1

Schematic diagram of miRNA detection methods based on rolling circle amplification (RCA). This material was reprinted (adapted) with permission from Ref. [19]. Copyright 2017 American Chemical Society.

Figure 2

Schematic diagram of miRNA detection methods based on duplex-specific nuclease signal amplification (DSNSA). The black circle represents the quencher group, the gray pentagram represents the quenched fluorophore, and the green glowing pentagram represents the fluorophore with restored fluorescence. This material was reprinted (adapted) with permission from Ref. [19]. Copyright 2017 American Chemical Society.

Figure 3

Schematic diagram of miRNA detection methods based on catalytic hairpin assembly (CHA). This material was reprinted (adapted) with permission from Ref. [19]. Copyright 2017 American Chemical Society.

Figure 4

Schematic diagram of miRNA detection methods based on strand-displacement amplification (SDA). The red dashed line represents the recognition site of the nicking endonuclease. This material was reprinted (adapted) with permission from Ref. [19]. Copyright 2017 American Chemical Society.

Figure 5

Schematic diagram of miRNA detection methods based on hybridization chain reaction (HCR). In primers HP1 and HP2, identical colors denote complementary base sequences. The miRNA indicated by the black line is capable of recognizing the yellow region in primer HP1. This material was reprinted (adapted) with permission from Ref. [19]. Copyright 2017 American Chemical Society.

Figure 6

Schematic diagram of miRNA detection methods based on loop-mediated isothermal amplification (LAMP). This material was reprinted (adapted) with permission from Ref. [19]. Copyright 2017 American Chemical Society.

Figure 7

Schematic diagram of miRNA detection methods based on the exponential amplification reaction (EXPAR) technique. The red dashed line represents the recognition site of the nicking endonuclease. This material was reprinted (adapted) with permission from Ref. [19]. Copyright 2017 American Chemical Society.

Figure 8

Mechanism of the SPHIA method for nucleic acid detection as depicted in a schematic diagram. This material was reprinted with permission from Ref. [58]. Copyright 2020 American Chemical Society.