Please use this identifier to cite or link to this item: http://ahro.austin.org.au/austinjspui/handle/1/9847
Title: Comparison of three PCR primer sets for identification of vanB gene carriage in feces and correlation with carriage of vancomycin-resistant enterococci: interference by vanB-containing anaerobic bacilli.
Authors: Ballard, Susan A;Grabsch, Elizabeth A;Johnson, Paul D R;Grayson, M Lindsay
Affiliation: Infectious Diseases Department, Austin Hospital, Austin Health, Studley Rd., Heidelberg, VIC 3084, Australia.
Issue Date: 1-Jan-2005
Citation: Antimicrobial Agents and Chemotherapy; 49(1): 77-81
Abstract: We assessed the sensitivities and specificities of three previously described PCR primers on enrichment broth cultures of feces for the accurate detection of fecal carriage of vancomycin-resistant enterococci (VRE). In addition, we investigated specimens that were vanB PCR positive but VRE culture negative for the presence of other vanB-containing pathogens. Feces from 59 patients (12 patients carrying vanB Enterococcus faecium strains and 47 patients negative for VRE carriage) were cultured for 36 h in aerobic brain heart infusion (BHI) broth, anaerobic BHI (AnO(2)BHI) broth, or aerobic Enterococcosel (EC) broth. DNA was extracted from the cultures and tested for the presence of vanB by using the PCR primers of Dutka-Malen et al. (S. Dutka-Malen, S. Evers, and P. Courvalin, J. Clin. Microbiol. 33:24-27, 1995), Bell et al. (J. M. Bell, J. C. Paton, and J. Turnidge, J. Clin. Microbiol. 36:2187-2190, 1998), and Stinear et al. (T. P. Stinear, D. C. Olden, P. D. R. Johnson, J. K. Davies, and M. L. Grayson, Lancet 357:855-856, 2001). The sensitivity (specificity) of PCR compared with the results of culture on BHI, AnO(2)BHI, and EC broths were 67% (96%), 50% (94%), and 17% (100%), respectively, with the primers of Dutka-Malen et al.; 92% (60%), 92% (45%), and 92% (83%), respectively, with the primers of Bell et al.; and 92% (49%), 92% (43%), and 100% (51%) respectively, with the primers of Stinear et al. The primers of both Bell et al. and Stinear et al. were significantly more sensitive than those of Dutka-Malen et al. in EC broth (P = 0.001 and P < 0.001, respectively). The poor specificities for all primer pairs were due in part to the isolation and identification of six anaerobic gram-positive bacilli, Clostridium hathewayi (n = 3), a Clostridium innocuum-like organism (n = 1), Clostridium bolteae (n = 1), and Ruminococcus lactaris-like (n = 1), from five fecal specimens that were vanB positive but VRE culture negative. All six organisms were demonstrated to contain a vanB gene identical to that of VRE. VanB-containing bowel anaerobes may result in false-positive interpretation of PCR-positive fecal enrichment cultures as VRE, regardless of the primers and protocols used.
Internal ID Number: 15616278
URI: http://ahro.austin.org.au/austinjspui/handle/1/9847
DOI: 10.1128/AAC.49.1.77-81.2005
URL: http://www.ncbi.nlm.nih.gov/pubmed/15616278
Type: Journal Article
Subjects: Bacteria, Anaerobic.genetics
Bacterial Proteins.genetics
Carrier State.microbiology
Culture Media
DNA Primers
Enterococcus.drug effects.genetics.isolation & purification
Enterococcus faecium.drug effects.genetics
Feces.microbiology
Gram-Positive Bacteria.drug effects.genetics
Gram-Positive Bacterial Infections.microbiology
Humans
Molecular Sequence Data
Polymerase Chain Reaction.methods
Sensitivity and Specificity
Sequence Analysis, DNA
Vancomycin Resistance.genetics
Appears in Collections:Journal articles

Files in This Item:
There are no files associated with this item.


Items in AHRO are protected by copyright, with all rights reserved, unless otherwise indicated.