Rapid identification of microorganisms by mass spectrometry: improved performance by incorporation of in-house spectral data into a commercial database.
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is increasingly used as a microbial diagnostic method for species identification of pathogens. However, MALDI-TOF identification of bacteria at the species level remains unsatisfactory, with the major problem being an incomplete database that still needs refinement and expansion. Augmentation of the original MALDI BioTyper 2.0 (Bruker) database by incorporating mass spectra obtained in-house from clinical isolates may increase the identification rate at the species level. We conducted a prospective study to assess whether the augmented database can improve the performance of MALDI-TOF MS for routine identification of species. Cluster analyses revealed distinct differences in MS spectral profiles of clinical isolates obtained in our hospital and those of ATCC strains in the Bruker database. In the first part of the study, which was performed over 3 weeks, 259 bacterial isolates were subjected to analysis by MALDI-TOF MS, and MS spectra of 229 successfully identified isolates (49 species) were incorporated into the original database to give the augmented Bruker-Chiba database. In a second separate analysis, the concordance of identification of 498 clinical isolates of the 49 species with conventional methods was 87.1% (434/498) with the commercial Bruker database and 98.0% (488/498) using the Bruker-Chiba database. These results indicate that refinement of a commercial database can be achieved relatively easy and effectively by incorporating MS spectra of clinical isolates obtained in a clinical laboratory.
Clinical Proteomics Research Center, Chiba University Hospital, 1-8-1 Inohana, Chiba City, Chiba 260-8670, Japan.
Kazuyuki
K
Sogawa
Masaharu
M
Watanabe
Kenichi
K
Sato
Syunsuke
S
Segawa
Akiko
A
Miyabe
Syota
S
Murata
Tomoko
T
Saito
Fumio
F
Nomura
D001419:Bacteria / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D001483:Base Sequence; D017931:DNA Primers; D016208:Databases, Factual; D010802:Phylogeny; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D017931:DNA Primers; D012336:RNA, Ribosomal, 16S