Novel specific peaks for differentiating the Lactobacillus plantarum group using matrix-assisted laser desorption ionization time-of-flight mass spectrometry

Abstract

Identifying the Lactobacillus plantarum group using conventional taxonomic methods such as biochemical analysis and 16S rRNA gene sequencing is inaccurate, expensive, and time-consuming. In this study, for the first time, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry was used to identify the L. plantarum group and develop a classification method for species level differentiation with specific peaks based on mass spectra. Furthermore, from the mass spectra of 131 isolates aligned with the biotyper database, 131 isolates (100%) were correctly identified at the species level with a mean score of 2.316. However, commercial databases could not accurately differentiate some isolates of L. plantarum group species because the same colony was identified as different species with similar score values. Moreover, these two species showed a similar mass pattern in the main spectrum profiles-dendrogram and Principal component analysis clustering generated by the mass peak of the reference strains and isolates. Specific peaks to each species were investigated from the analyzed mass peak, and they clearly showed that three species could be differentiated. These peaks were verified by re-identifying 131 isolates, and it demonstrated 100% specificity and accuracy. Also, using a specific peak, isolates that were undifferentiated from the biotyper database were clearly identified as one species, similar to species-specific polymerase chain reaction. Our data demonstrate that the specific peaks accurately differentiate the L. plantarum group and enable high-resolution identification at the species level; this methodology can be used to rapidly and easily identify them and determine their nomenclature.

Keywords: Identification; Lactobacillus plantarum group; MALDI-TOF MS; Specific PCR; Specific peak; bioTyper database.