The World of Nucleic Acid Based Mass Spectrometry for Microbial and Viral Detection
Christiane Honisch
from: MALDI-TOF Mass Spectrometry in Microbiology (Edited by: Markus Kostrzewa and Sören Schubert). Caister Academic Press, U.K. (2016) Pages: 141-156.
Abstract
Matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF MS) has been applied to nucleic acid based research and specifically sequencing for over 25 years with increasing clinical use today. In contrast to whole cell MS, where cultured material is the source for the identification of a microbial sample based on a proteomic fingerprint, nucleic acid MS based microbial detection, identification and characterization uses PCR amplifications of specific genomic target regions of interest followed by the detection of single nucleotide polymorphisms via primer extension reactions or followed by comparative sequence analysis of mass peak pattern generated via in vitro transcription and base-specific cleavage. Applications include the detection and identification of both microbial and viral pathogens, tracking of transmissions and the characterization of genetic heterogeneity by variant analysis. Multiplexing levels of up to 40- 60 targets per reaction surpass real-time PCR applications, but fall short when compared to the potential of next-generation sequencing. The unique feature of mass spectrometry (MS) though is the detection of various biomarker molecules including proteins, lipids, small molecules, carbohydrates and nucleic acids on one single platform read more ...
