Table 1 Conventional miRNA detection methods
Detection method | Advantages | Disadvantages | References |
|---|---|---|---|
Northern blotting | -High specificity-Readily available and easy-to-perform | -Low-throughput-Low sensitivity-Laborious and very time consuming | |
In situ hybridization (ISH) with locked nucleic acid probes | -Able to monitor the cellular and sub-cellular distributions and to determine the spatiotemporal expression profile of miRNAs | -Low-throughput-Semi-quantitative analysis of miRNA expression | [90] |
Reverse transcription qPCR (RT-qPCR) | -High sensitivity and specificity -Suitable for measuring smaller miRNA panels-Can be used for absolute quantification-Easy to incorporate into the workflow for laboratories that are used to qPCR-Suitable for routine measurements in laboratory settings-Can be automated to a high degree | -Cannot identify novel miRNAs -Medium-throughput with respect to the number of samples that can be processed per day -Optimal reaction conditions may differ considerably between miRNAs due to sequence-specific differences in primer annealing | |
Microarray | -Rather low-cost and high-throughput considering the number of samples that can be processed per day -Suitable for comparing the relative abundance of specific miRNAs between two states (e.g., ‘diseased’ versus ‘non- diseased) | -Low sensitivity and specificity -Long turnaround time-Poor degree of automation-Not suitable for absolute quantification-Cannot identify novel miRNAs | |
Next generation sequencing (NGS) | -Widest range of applications -Very high sensitivity-High accuracy in distinguishing variants of miRNAs that are very similar in sequence | -Substantial computational support needed for data analysis-Cannot be used for absolute quantification-Rather expensive and not available everywhere-Not so suitable for high-throughput laboratory settings yet |
