title: Multiplex identification of gram-positive bacteria and resistance determinants directly from positive blood culture broths: evaluation of an automated microarray-based nucleic acid test.
abstract: BACKGROUND
  A multicenter study was conducted to evaluate the diagnostic accuracy (sensitivity and specificity) of the Verigene Gram-Positive Blood Culture Test (BC-GP) test to identify 12 Gram-positive bacterial gene targets and three genetic resistance determinants directly from positive blood culture broths containing Gram-positive bacteria.
  
  
  RESULTS
  1,252 blood cultures containing Gram-positive bacteria were prospectively collected and tested at five clinical centers between April, 2011 and January, 2012. An additional 387 contrived blood cultures containing uncommon targets (e.g., Listeria spp., S. lugdunensis, vanB-positive Enterococci) were included to fully evaluate the performance of the BC-GP test. Sensitivity and specificity for the 12 specific genus or species targets identified by the BC-GP test ranged from 92.6%-100% and 95.4%-100%, respectively. Identification of the mecA gene in 599 cultures containing S. aureus or S. epidermidis was 98.6% sensitive and 94.3% specific compared to cefoxitin disk method. Identification of the vanA gene in 81 cultures containing Enterococcus faecium or E. faecalis was 100% sensitive and specific. Approximately 7.5% (87/1,157) of single-organism cultures contained Gram-positive bacteria not present on the BC-GP test panel. In 95 cultures containing multiple organisms the BC-GP test was in 71.6% (68/95) agreement with culture results. Retrospective analysis of 107 separate blood cultures demonstrated that identification of methicillin resistant S. aureus and vancomycin resistant Enterococcus spp. was completed an average of 41.8 to 42.4 h earlier using the BC-GP test compared to routine culture methods. The BC-GP test was unable to assign mecA to a specific organism in cultures containing more than one Staphylococcus isolate and does not identify common blood culture contaminants such as Micrococcus, Corynebacterium, and Bacillus.
  
  
  CONCLUSIONS
  The BC-GP test is a multiplex test capable of detecting most leading causes of Gram-positive bacterial blood stream infections as well as genetic markers of methicillin and vancomycin resistance directly from positive blood cultures.
authors.affiliation: Department of Pathology, Medical College of Wisconsin, Milwaukee, Wisconsin, United States of America.
authors.forename: Blake W
authors.initials: BW
authors.lastname: Buchan
authors.forename: Christine C
authors.initials: CC
authors.lastname: Ginocchio
authors.forename: Ryhana
authors.initials: R
authors.lastname: Manii
authors.forename: Robert
authors.initials: R
authors.lastname: Cavagnolo
authors.forename: Preeti
authors.initials: P
authors.lastname: Pancholi
authors.forename: Lettie
authors.initials: L
authors.lastname: Swyers
authors.forename: Richard B
authors.initials: RB
authors.lastname: Thomson
authors.forename: Christopher
authors.initials: C
authors.lastname: Anderson
authors.forename: Karen
authors.initials: K
authors.lastname: Kaul
authors.forename: Nathan A
authors.initials: NA
authors.lastname: Ledeboer
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001769:Blood / Q000382:microbiology; D015169:Colony Count, Microbial / Q000379:methods*; D024881:Drug Resistance, Bacterial / Q000187:drug effects*; D005819:Genetic Markers / Q000187:drug effects; D005840:Gentian Violet / Q000378:metabolism; D006094:Gram-Positive Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D046228:Microarray Analysis / Q000379:methods*; D010619:Phenazines / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D014481:United States
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D005819:Genetic Markers; C035863:Gram's stain; D010619:Phenazines; D005840:Gentian Violet