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Identification of Rare Bacterial Pathogens by 16S rRNA Gene Sequencing and MALDI-TOF MS
Published on: July 11, 2016
PVBase: A MALDI-TOF MS Database for Fast Identification and Characterization of Potentially Pathogenic Vibrio Species
Tingting Liu1,2, Lin Kang2, Jinglin Xu1,2
1Hebei Key Laboratory of Animal Physiology, Biochemistry and Molecular Biology, College of Life Sciences, Hebei Normal University, Shijiazhuang, China.
Abstract:
The potentially pathogenic species of the genus Vibrio pose a threat to both humans and animals, creating medical burdens and economic losses to the mariculture industry. Improvements in surveillance and diagnosis are needed to successfully manage vibriosis outbreaks. Matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) can provide rapid diagnosis and has been widely used in the identification of Vibrio spp. The main weakness of this technology is the limited number of strains and species of Vibrio in the existing commercial database. Here, we develop a new in-house database named PVBase containing 790 main spectra projections (MSP) of ten Vibrio species that come from various regions of China and include abundant clinical and environmental strains. PVBase was validated through a blind test of 65 Vibrio strains. The identification accuracy and scoring of Vibrio strains was greatly improved through the addition of PVBase. Identification accuracy increased from 73.4 to 100%. The number of strains with identification scores above 2.2 increased from 53.1% to 96.9% and 53.1% of strains had an identification score above 2.59. Moreover, perfect discrimination was obtained when using all of the MSPs created for the Vibrio species, even for very closely related species such as V. cholerae, V. albensis, and V. mimicus or V. alginolyticus, V. parahaemolyticus, and V. harveyi. In addition, we used phyloproteomic analysis to study whether there are differences in protein fingerprints of different regions or pathogenic strains. We found that MSP characteristics of Vibrio species were not related to their region or source. With the construction of PVBase, the identification efficiency of potentially pathogenic Vibrio species has been greatly improved, which is an important advance for epidemic prevention and control, and aquaculture disease detection.
Insights
A new database, PVBase, significantly enhances the identification of pathogenic Vibrio species using MALDI-TOF MS. This improves disease surveillance and aquaculture diagnostics, increasing accuracy from 73.4% to 100%.
Area of Science:
- Microbiology
- Biotechnology
- Veterinary Medicine
Background:
- Potentially pathogenic Vibrio species cause significant human and animal health issues, leading to economic losses in mariculture.
- Effective surveillance and diagnosis are crucial for managing vibriosis outbreaks.
- Matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) offers rapid bacterial identification but is limited by incomplete databases.
Purpose of the Study:
- To develop and validate a comprehensive in-house database (PVBase) for improved identification of Vibrio species.
- To enhance the accuracy and efficiency of MALDI-TOF MS for Vibrio identification in clinical and environmental samples.
- To assess the utility of phyloproteomic analysis for understanding Vibrio strain variations.
Main Methods:
- Creation of PVBase containing 790 main spectra projections (MSP) from ten Vibrio species across various Chinese regions.
- Validation of PVBase through a blind test involving 65 Vibrio strains.
- Phyloproteomic analysis to investigate protein fingerprint variations related to geographic origin or pathogenicity.
Main Results:
- PVBase addition increased Vibrio identification accuracy from 73.4% to 100%.
- The percentage of strains with identification scores above 2.2 rose from 53.1% to 96.9%.
- Perfect discrimination was achieved for closely related Vibrio species, and MSP characteristics were independent of region or source.
Conclusions:
- The PVBase database significantly improves the identification efficiency of potentially pathogenic Vibrio species.
- This advancement is critical for epidemic prevention, disease control, and aquaculture health monitoring.
- MALDI-TOF MS, augmented by PVBase, provides a powerful tool for rapid and accurate Vibrio diagnostics.
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