Nucleic acid mass spectrometry

Nucleic acid mass spectrometry is a multiplex PCR-based detection system developed on the foundation of MALDI-TOF MS mass spectrometry. Nucleic acid mass spectrometry primarily relies on multiplex PCR, high-throughput chips, and time-of-flight mass spectrometry to perform nucleic acid detection. This technology enables simultaneous detection of dozens or even hundreds of target markers from a single sample, with the capacity to process over 1,000 samples per day.

The main steps of nucleic acid mass spectrometry:

1. Polymerase Chain Reaction (PCR): Design one forward primer and one reverse primer targeting the locus to be detected. Using the sample DNA as a template, perform PCR to obtain a PCR product fragment containing the target locus.

2. Treatment with shrimp alkaline phosphatase (SAP): Add SAP (10322ES) to the PCR product to perform a dephosphorylation reaction, thereby removing any residual dNTPs from the reaction system.

3. Single-base extension reaction: Add a single-base extension primer and ddNTPs to perform the single-base extension reaction.

4. Resin Purification: Add desalting resin to the extended product for desalting and purification, thereby preventing salt ion peaks from interfering with the mass spectrometry results.

5. Sample-matrix binding: The sample is spotted onto a chip coated with a matrix, and then subjected to mass spectrometry analysis. Different gene fragments ultimately generate distinct detection peak profiles.

In recent years, nucleic acid mass spectrometry has emerged as a new molecular diagnostic platform, joining PCR and NGS as a key technology in the field. While fluorescence-based quantitative PCR offers rapid detection speeds but has limited throughput, making it difficult to efficiently meet the demand for analyzing dozens to hundreds of sites across multiple genes, high-throughput sequencing, though boasting extremely high throughput, comes with high testing costs, long project turnaround times, and requires specialized analysis and interpretation of data—thus presenting a relatively high technical barrier. In contrast, nucleic acid mass spectrometry delivers stable and accurate results at lower costs, perfectly addressing the need for qualitative and quantitative detection of SNPs and gene mutations at moderate throughputs. Moreover, it enables convenient and rapid detection of DNA methylation and copy number variations (CNVs), further highlighting its strong competitive edge in the field of genetic testing and establishing it as one of the rapidly growing mainstays in life sciences, particularly in clinical diagnostics.

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