# RNA Modification Related Diseases and Sensing Methods
## 5 Detection Methods for RNA Modifications  
### Quantitative Methods for Modified Bases  
To accurately examine and profile RNA modifications, various challenging methods must be used due to the differences in the position and splicing of adducts as well as the length and structure of the nucleotide strand [2; 82]. Several methods have been reported for measuring base modification levels, including dot blotting [83], Northern blotting with antibodies [84], Maz RNA endonuclease [85], high-resolution melting (HRM) [72], RNA photo-crosslinkers and quantitative proteomics (PCL-Proteomics) [86], and silver SiO2-based electrochemical immunosensors (ECI) [87].  
RNA modifications are chemical alterations to RNA bases that can occur in various types of RNA molecules. These modifications can regulate RNA structure, stability, and function and play crucial roles in gene expression and disease development. Therefore, detecting and analyzing RNA modifications has become a significant area of research in molecular biology and biomedicine. Several techniques have been developed to detect RNA modifications, including mass spectrometry, high-performance liquid chromatography (HPLC), antibody-based assays, and next-generation sequencing (NGS). Mass spectrometry is a powerful tool for identifying and quantifying RNA modifications with high accuracy and resolution. However, it requires sophisticated instrumentation, sample preparation, and expertise, and it is not suitable for high-throughput analysis [88]. HPLC is another widely used technique for detecting RNA modifications based on the separation and quantification of modified nucleosides using a chromatographic column. HPLC has high sensitivity and specificity, but it is time-consuming, requires specialized equipment, and may not be able to distinguish between closely related modifications [35]. Antibody-based assays, such as enzyme-linked immunosorbent assay (ELISA) and dot blot, use specific antibodies to recognize and quantify RNA modifications. These methods are relatively simple, fast, and cost-effective, but they have limited sensitivity and specificity and may cross-react with other modifications or non-modified nucleotides [89]. NGS-based methods, such as m6A-seq and miCLIP, are emerging as powerful tools for detecting and mapping RNA modifications at a transcriptome-wide scale. These methods use RNA sequencing and specific probes or antibodies to enrich and sequence modified RNA molecules. NGS has high sensitivity, specificity, and throughput, but it requires complex data analysis and may generate false-positive signals due to technical artifacts or sequencing errors [90]. Other emerging techniques for detecting RNA modifications include nanopore sequencing, single-molecule imaging, and chemical probing [91]. These methods have their advantages and limitations and are still under development. In conclusion, the choice of RNA modification detection method depends on the research question, the available resources, and the desired accuracy, sensitivity, and throughput. Therefore, it is essential to evaluate the performance and limitations of each technique carefully and to validate the results using complementary methods. Moreover, methods to determine both the position and the number of base alterations have been published [2; 4; 92; 93; 94; 95; 96].