Table 4.

An overview of different methods for detecting miRNAs

Detection Method	Description	Advantages	Limitations	References
Quantitative PCR (qPCR)	Measures miRNA levels using specific primers and probes, providing quantitative data	High sensitivity and specificity; widely used and validated	It requires prior RNA extraction and is limited to known miRNAs	[183]
Microarray Analysis	It utilizes a chip with probes for multiple miRNAs to profile their expression levels across various samples	It can detect multiple miRNAs simultaneously, making it suitable for broad profiling	Less sensitive for low-abundance miRNAs; complex data analysis	[184]
Next-Generation Sequencing (NGS)	Provides comprehensive profiling of miRNAs by sequencing RNA fragments, allowing for the discovery of novel miRNAs	High throughput; can identify novel miRNAs; provides detailed expression profiles	High cost; complex data analysis; requires significant bioinformatics resources	[185]
Northern blotting	Detects miRNAs based on their size and abundance using gel electrophoresis followed by hybridization with a labeled probe	It provides size information and is suitable for the specific detection of miRNA	Time consuming; requires large amounts of RNA; less quantitative	[187]
In Situ Hybridization (ISH)	Detects miRNA expression in tissue samples using labeled probes that bind to complementary miRNA sequences	Allows localization of miRNA within tissue; provides spatial context	Less quantitative; requires tissue samples	[188]
RNA Sequencing (RNA-seq)	A form of NGS focused on sequencing all RNA species, including miRNAs, to analyze their expression and discover novel miRNAs	Comprehensive; detects known and novel miRNAs; high resolution	Expensive; requires bioinformatics expertise; data complexity	[186]
Droplet Digital PCR (ddPCR)	Quantifies miRNAs with high precision by partitioning the sample into droplets and performing PCR in each droplet	Highly sensitive and precise; allows absolute quantification	Expensive equipment; limited multiplexing capabilities	[189]
Biosensors	Employs various sensing technologies (e.g., electrochemical, optical) to detect specific miRNA sequences in real time	It can be susceptible and enables real-time monitoring	It may require extensive calibration, which is less common in standard laboratories	[190]