title: Comparison of MIDI Sherlock system and pulsed-field gel electrophoresis in characterizing strains of methicillin-resistant Staphylococcus aureus from a recent hospital outbreak.
abstract: An outbreak of methicillin-resistant Staphylococcus aureus infections at the University of Utah Health Sciences Center occurred over a 7-month period. While the isolates phenotypically appeared to be similar in gross morphology and have similar Vitek antibiotic susceptibility patterns, two additional methods of strain characterization were evaluated to enhance the epidemiological investigation: pulsed-field gel electrophoresis and gas chromatography with the MIDI Sherlock system. Sherlock uses gas chromatography to qualitatively and quantitatively analyze the cellular fatty acid composition of organisms and creates two-dimensional plots based on principal-component analysis to define groups of closely related organisms. All isolates were also evaluated by digesting their chromosomal DNAs with the low-frequency-cutting enzyme SmaI and separating the restriction fragments by contour-clamped homogeneous electric field gel electrophoresis. Sample preparation for this pulsed-field gel electrophoresis included a novel cell lysis procedure involving achromopeptidase, greatly reducing the turnaround time. Isolates tested were recovered from the following: 45 suspected outbreak patients, 6 hospitalized patients believed to be unrelated to the outbreak, 6 patients from outside the hospital, and one health care practitioner implicated in the outbreak. Of 45 phenotypically similar suspect strains, 43 clustered tightly on the Sherlock two-dimensional plot. All outbreak patient isolates were also identical by pulsed-field gel electrophoresis with the exception of the same two outliers identified by Sherlock. In this epidemiologic investigation, we found an excellent correlation between the Sherlock and pulsed-field gel electrophoresis results for strain characterization of methicillin-resistant S. aureus.
authors.affiliation: Associated Regional and University Pathologists, Inc., Salt Lake City, Utah 84108, USA.
authors.forename: R B
authors.initials: RB
authors.lastname: Leonard
authors.forename: J
authors.initials: J
authors.lastname: Mayer
authors.forename: M
authors.initials: M
authors.lastname: Sasser
authors.forename: M L
authors.initials: ML
authors.lastname: Woods
authors.forename: B R
authors.initials: BR
authors.lastname: Mooney
authors.forename: B G
authors.initials: BG
authors.lastname: Brinton
authors.forename: P L
authors.initials: PL
authors.lastname: Newcomb-Gayman
authors.forename: K C
authors.initials: KC
authors.lastname: Carroll
mesh_terms: D015373:Bacterial Typing Techniques; D002849:Chromatography, Gas; D003428:Cross Infection; D004269:DNA, Bacterial / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific; D004196:Disease Outbreaks; D016521:Electrophoresis, Gel, Pulsed-Field; D005227:Fatty Acids / Q000032:analysis; D014644:Genetic Variation; D006761:Hospitals; D006801:Humans; D016106:Methicillin Resistance; D008826:Microbial Sensitivity Tests; D013203:Staphylococcal Infections / Q000453:epidemiology* / Q000517:prevention & control; D013211:Staphylococcus aureus / Q000737:chemistry / Q000145:classification* / Q000235:genetics; D014589:Utah / Q000453:epidemiology
chemical_list: D004269:DNA, Bacterial; D005227:Fatty Acids; D015252:Deoxyribonucleases, Type II Site-Specific