Table 4.
RNA detection methods
| Technology | Mechanisms | Advantage | Disadvantage | Reference |
|---|---|---|---|---|
| RNA fluorescence in situ hybridization (RNA-FISH) | Hybridization signals were observed using fluorescence microscopy after binding to the target RNA with a fluorescent probe complementary to the target RNA sequence | High sensitivity and specificity, multi-color detection, relatively simple and time-consuming operation, tissue morphology can be maintained for detection | High sample requirements, need to ensure RNA integrity, need specialized equipment and probes, high cost, limited accuracy of quantification | 102 |
| RT-PCR (reverse transcription PCR) | PCR amplification after reverse transcription of RNA to cDNA | It is highly sensitive and specific, suitable for the detection of a wide range of RNAs, less time consuming and more accurate. | Complexity of operation, susceptibility to contamination by foreign products, expensive equipment and reagents | 103 |
| Northern Blotting | Complexity of operation, susceptibility to contamination by foreign products, expensive equipment and reagents | High sensitivity and specificity, quantitative detection of RNA compared to RNA-FISH | Time-consuming, more complex operations, high sample requirements, need to ensure RNA integrity, need specialized equipment and probes, higher costs | 104 |
| in situ hybridization | The principle is similar to RNA-FISH, but labeled using markers such as radioisotopes, biotin, digoxin, etc., and finally visualized by radioactive autoradiography, immunohistochemistry, etc. | Both DNA and RNA can be detected at a moderate cost | Not as accurate as RNA-FISH, multiple hybridizations are not as simple as RNA-FISH, can only capture RNA from cells at a certain time point | 105 |
| RNA microarray | Hybridization of RNA by immobilizing a large number of probes on a microarray | High throughput, accurate quantification and good reproducibility. | Can only detect highly expressed RNAs and cannot cover the full range of RNAs, especially lncRNAs. cost is high and affected by experimental complexity. | 106 |
| RNA sequencing | Direct sequencing of RNA molecules using high-throughput sequencing technology | Detects all RNAs, capable of deep sequencing with high sensitivity and specificity | Costly, requires advance removal of rRNAs | 106 |