One-by-one single-molecule counting method for digital quantification of SARS-CoV-2 RNA

Area of Science:

• Biotechnology
• Molecular Biology
• Nanotechnology

Background:

• Accurate digital counting of individual nucleic acid molecules is crucial for biological research and genetic disease diagnostics.
• Existing single-molecule detection technologies face limitations in achieving precise quantification.

Purpose of the Study:

• To develop a novel one-by-one single-molecule counting method for digital quantification of SARS-CoV-2 RNA.
• To demonstrate the capability of discriminating single-base mutations in target RNAs.

Main Methods:

• Utilizing fluorescent micromotors functionalized with peptide nucleic acids (PNAs) for specific capture of single target RNA molecules.
• Employing electric fields to propel RNA-micromotors to target districts for accurate counting.
• Developing a digital quantification approach for single nucleic acid molecules.

Main Results:

• Successfully demonstrated one-by-one counting of individual SARS-CoV-2 RNA molecules.
• Achieved accurate digital quantification of target RNA molecules.
• Showcased the method's ability to discriminate single-base mutations in RNA.

Conclusions:

• The developed method provides a novel approach for precise digital quantification of nucleic acid molecules.
• This technique holds significant potential for clinical diagnostics, including genetic disease detection and virus traceability surveys.
• The ability to detect single-base mutations enhances its utility for molecular diagnostics and epidemiology.