title: Application of Routine Diagnostic Procedure, VITEK 2 Compact, MALDI-TOF MS, and PCR Assays in Identification Procedure of Bacterial Strain with Ambiguous Phenotype.
abstract: In diagnostic microbiology as well as in microbiological research, the identification of a microorganism is a crucial and decisive stage. A broad choice of methods is available, based on both phenotypic and molecular properties of microbes. The aim of this study was to compare the application of phenotypic and molecular tools in bacterial identification on the example of Gram-negative intestine rod with an ambiguous phenotype. Different methods of identification procedure, which based on various properties of bacteria, were applied, e.g., microscopic observation of single-bacterial cells, macroscopic observation of bacterial colonies morphology, the automated system of microorganism identification (biochemical tests), the mass spectrometry method (analysis of bacterial proteome), and genetic analysis with PCR reactions. The obtained results revealed discrepancies in the identification of the tested bacterial strain with an atypical phenotype: mucous morphology of colonies, not characteristic for either E. coli and Citrobacter spp., mass spectrometry analysis of proteome initially assigned the tested strain to Citrobacter genus (C. freundii) and biochemical profiles pointed to Escherichia coli. A decisive method in the current study was genetic analysis with PCR reactions which identified conserved genetic sequences highly specific to E. coli species in the genome of the tested strain.
authors.affiliation: Department of Microbiology, Institute of Genetics and Microbiology, University of Wrocław, Przybyszewskiego 63/77, 51-148, Wrocław, Poland. marta.ksiazczyk@uwr.edu.pl.
authors.forename: Marta
authors.initials: M
authors.lastname: Książczyk
authors.affiliation: Department of Epizootiology and Clinic of Bird and Exotic Animals, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, Pl. Grunwaldzki 45, 50-366, Wrocław, Poland.
authors.forename: Maciej
authors.initials: M
authors.lastname: Kuczkowski
authors.affiliation: Department of Microbiology, Institute of Genetics and Microbiology, University of Wrocław, Przybyszewskiego 63/77, 51-148, Wrocław, Poland.
authors.forename: Bartłomiej
authors.initials: B
authors.lastname: Dudek
authors.affiliation: Department of Microbiology, Institute of Genetics and Microbiology, University of Wrocław, Przybyszewskiego 63/77, 51-148, Wrocław, Poland.
authors.forename: Kamila
authors.initials: K
authors.lastname: Korzekwa
authors.affiliation: Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Rudolfa Weigla 12, 53-114, Wrocław, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Tobiasz
authors.affiliation: Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Rudolfa Weigla 12, 53-114, Wrocław, Poland.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Korzeniowska-Kowal
authors.affiliation: Department of Microbiology, Institute of Genetics and Microbiology, University of Wrocław, Przybyszewskiego 63/77, 51-148, Wrocław, Poland.|Department Physicochemistry of Microorganisms, Institute of Genetics and Microbiology, University of Wrocław, Przybyszewskiego 63/77, 51-148, Wrocław, Poland.
authors.forename: Emil
authors.initials: E
authors.lastname: Paluch
authors.affiliation: Department of Epizootiology and Clinic of Bird and Exotic Animals, Faculty of Veterinary Medicine, Wrocław University of Environmental and Life Sciences, Pl. Grunwaldzki 45, 50-366, Wrocław, Poland.
authors.forename: Alina
authors.initials: A
authors.lastname: Wieliczko
authors.affiliation: Department of Microbiology, Institute of Genetics and Microbiology, University of Wrocław, Przybyszewskiego 63/77, 51-148, Wrocław, Poland.
authors.forename: Gabriela
authors.initials: G
authors.lastname: Bugla-Płoskońska
mesh_terms: D000818:Animals; D015373:Bacterial Typing Techniques / Q000295:instrumentation / Q000379:methods*; D005243:Feces / Q000382:microbiology; D006090:Gram-Negative Bacteria / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016905:Gram-Negative Bacterial Infections / Q000175:diagnosis / Q000382:microbiology*; D006801:Humans; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*