Research on the establishment and application of protein fingerprint spectrum database of Burkholderia pseudomallei in Hainan Province China

Abstract

Burkholderia pseudomallei (B. pseudomallei) is the etiologic agent of melioidosis. The aim of this study was to establish a SuperSpectrum of B. pseudomallei in Hainan and evaluate its application value in the rapid identification of clinical isolates of B. pseudomallei. In total, 99 isolates of B. pseudomallei were obtained between 2010-2019 in different regions of Hainan Province, multilocus sequence typing (MLST) was performed, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was applied for spectrum acquisition. A SuperSpectrum was created based on the selection of 80 representative average spectra for 17 isolates of B. pseudomallei. Then we validated the SuperSpectra with 82 strains of B. pseudomallei, eight strains of Burkholderia thailandensis, two strains of Burkholderia cepacia, one strain of Burkholderia cenocepacia and one strain of Burkholderia multivorans, as well as one strain of Burkholderia gladioli identified by MLST typing, 16S rRNA gene sequencing and Vitek 2 Compact. The results showed that all samples in this study were confirmed at the species level. Protein fingerprints spectra showed that specific peaks occurred in B. pseudomallei from the Hainan region. The result of clustering typing indicated that B. pseudomallei and its closely related species could be well classified by MALDI-TOF MS at the protein level.

Keywords

• 16S rRNA gene sequencing analysis
• Burkholderia pseudomallei
• MALDI-TOF MS
• superspectra
• vitek 2 compact