title: A new assay for the simultaneous identification and differentiation of Klebsiella oxytoca strains.
abstract: Klebsiella oxytoca is the second most frequently identified species of Klebsiella isolated from hospitalized patients. Klebsiella spp. is difficult to identify using conventional methods and is often misclassified in clinical microbiology laboratories. K. oxytoca is responsible for an increasing number of multi-resistant infections in hospitals because of insufficient detection and identification. In this study, we propose a new simple method called pehX-LM PCR/XbaI, which simultaneously indicates K. oxytoca species and genotype by the fingerprint pattern. The pehX-LM PCR/XbaI is a combination of the following two methods: species-specific amplification of pehX gene and non-specific amplification of short restriction fragments by the LM PCR method. The specificity and the discrimination power of the pehX-LM PCR/XbaI method were determined by typing 209 K. oxytoca strains (included 9 reference strains), 28 K. pneumoniae, and other 25 strains belonging to the Enterobacteriaceae. The typing results were confirmed by the PCR melting profile method. Unlike the known fingerprinting methods, the pehX-LM PCR/XbaI leads to a clear pattern (approx. 3-5 bands) with a sufficient, relatively high discriminatory power. As a result, the time and cost of a single analysis are lower. The method can be used both in clinical and environmental research.
authors.affiliation: Department of Molecular Biotechnology and Microbiology, Gdańsk University of Technology, ul. G. Narutowicza 11/12, 80-233, Gdańsk, Poland.
authors.forename: Karolina
authors.initials: K
authors.lastname: Stojowska-Swędrzyńska
authors.affiliation: Department of Molecular Biotechnology and Microbiology, Gdańsk University of Technology, ul. G. Narutowicza 11/12, 80-233, Gdańsk, Poland. beakrawc@pg.gda.pl.
authors.forename: Beata
authors.initials: B
authors.lastname: Krawczyk
mesh_terms: D006801:Humans; D007710:Klebsiella Infections / Q000175:diagnosis*; D041121:Klebsiella oxytoca / Q000145:classification* / Q000302:isolation & purification*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D058889:Molecular Typing / Q000379:methods*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012680:Sensitivity and Specificity
keywords: Enterobacteriaceae; Klebsiella oxytoca; Klebsiella spp.; LM PCR typing methods; PCR; pehX gene