Extended-spectrum β-lactamases (ESBLs) are emerging worldwide, making rapid and adequate ESBL detection crucial for infection control measures as well as for the choice of correct antimicrobial therapy. The aim of this study was to compare the performance of a novel rapid ligation-mediated real-time PCR (LM-PCR) with a combination disc test (CDT). In total, 172 prospective putative ESBL-positive Enterobacteriaceae isolates from clinical specimens based on VITEK2 results were included in this study and tested with the phenotypic CDT and the LM-PCR. Positive ESBL results were obtained in 100 and 95 isolates using CDT and LM-PCR, respectively. The sensitivity, specificity, negative predictive value and positive predictive value of the LM-PCR were 99.0, 92.2, 98.6 and 94.0 %, respectively, compared with the CDT. The LM-PCR technique provides an important reduction in turnaround time (~4.5 h versus overnight incubation using CDT) for ESBL confirmation. As a consequence, all ESBL results are available within the same day, making this assay an important tool for rapid and accurate ESBL detection.
BeceiroA., MaharjanS., GaultonT., DoumithM., SoaresN. C., DhanjiH., WarnerM., DoyleM., HickeyM.other authors2011; False extended-spectrum β-lactamase phenotype in clinical isolates of Escherichia coli associated with increased expression of OXA-1 or TEM-1 penicillinases and loss of porins. J Antimicrob Chemother 66:2006–2010 [View Article][PubMed]
BradfordP. A.2001; Extended-spectrum β-lactamases in the 21st century: characterization, epidemiology, and detection of this important resistance threat. Clin Microbiol Rev 14:933–951 [View Article][PubMed]
CarterM. W., OaktonK. J., WarnerM., LivermoreD. M.2000; Detection of extended-spectrum β-lactamases in klebsiellae with the Oxoid combination disk method. J Clin Microbiol 38:4228–4232[PubMed]
Cohen StuartJ., DierikxC., Al NaiemiN., KarczmarekA., Van HoekA. H. A. M., VosP., FluitA. C., ScharringaJ., DuimB.other authors2010; Rapid detection of TEM, SHV and CTX-M extended-spectrum β-lactamases in Enterobacteriaceae using ligation-mediated amplification with microarray analysis. J Antimicrob Chemother 65:1377–1381 [View Article][PubMed]
CuzonG., NaasT., BogaertsP., GlupczynskiY., NordmannP.2012; Evaluation of a DNA microarray for the rapid detection of extended-spectrum β-lactamases (TEM, SHV and CTX-M), plasmid-mediated cephalosporinases (CMY-2-like, DHA, FOX, ACC-1, ACT/MIR and CMY-1-like/MOX) and carbapenemases (KPC, OXA-48, VIM, IMP and NDM). J Antimicrob Chemother 67:1865–1869 [View Article][PubMed]
DallenneC., Da CostaA., DecréD., FavierC., ArletG.2010; Development of a set of multiplex PCR assays for the detection of genes encoding important β-lactamases in Enterobacteriaceae. J Antimicrob Chemother 65:490–495 [View Article][PubMed]
EndimianiA., HujerA. M., HujerK. M., GattaJ. A., SchriverA. C., JacobsM. R., RiceL. B., BonomoR. A.2010; Evaluation of a commercial microarray system for detection of SHV-, TEM-, CTX-M-, and KPC-type β-lactamase genes in Gram-negative isolates. J Clin Microbiol 48:2618–2622 [View Article][PubMed]
GarrecH., Drieux-RouzetL., GolmardJ.-L., JarlierV., RobertJ.2011; Comparison of nine phenotypic methods for detection of extended-spectrum β-lactamase production by Enterobacteriaceae. J Clin Microbiol 49:1048–1057 [View Article][PubMed]
Kluytmans-VandenberghM. F. Q., KluytmansJ. A. J. W., VossA.2005; Dutch guideline for preventing nosocomial transmission of highly resistant microorganisms (HRMO). Infection 33:309–313 [View Article][PubMed]
KohnerP. C., RobbertsF. J. L., CockerillF. R.III, PatelR.2009; Cephalosporin MIC distribution of extended-spectrum-β-lactamase- and pAmpC-producing Escherichia coli and Klebsiella species. J Clin Microbiol 47:2419–2425 [View Article][PubMed]
NijhuisR. H. T., van ZwetA. A., SavelkoulP. H. M., RooversE. A., BosboomR. W., PostmaB., van GriethuysenA. J.2011; Distribution of extended-spectrum β-lactamase genes using a commercial DNA micro-array system. J Hosp Infect 79:349–353 [View Article][PubMed]
NijssenS., FlorijnA., BontenM. J. M., SchmitzF. J., VerhoefJ., FluitA. C.2004; β-Lactam susceptibilities and prevalence of ESBL-producing isolates among more than 5000 European Enterobacteriaceae isolates. Int J Antimicrob Agents 24:585–591 [View Article][PubMed]
QueenanA. M., FolenoB., GownleyC., WiraE., BushK.2004; Effects of inoculum and β-lactamase activity in AmpC- and extended-spectrum β-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae clinical isolates tested by using NCCLS ESBL methodology. J Clin Microbiol 42:269–275 [View Article][PubMed]
van DoornumG. J. J., GuldemeesterJ., OsterhausA. D. M. E., NiestersH. G. M.2003; Diagnosing herpesvirus infections by real-time amplification and rapid culture. J Clin Microbiol 41:576–580 [View Article][PubMed]
VoetsG. M., FluitA. C., ScharringaJ., Cohen StuartJ., Leverstein-van HallM. A.2011; A set of multiplex PCRs for genotypic detection of extended-spectrum β-lactamases, carbapenemases, plasmid-mediated AmpC β-lactamases and OXA β-lactamases. Int J Antimicrob Agents 37:356–359 [View Article][PubMed]
WassenbergM. W. M., KluytmansJ. A., BoxA. T. A., BosboomR. W., BuitingA. G. M., van ElzakkerE. P. M., MelchersW. J. G., van RijenM. M. L., ThijsenS. F. T.other authors2010; Rapid screening of methicillin-resistant Staphylococcus aureus using PCR and chromogenic agar: a prospective study to evaluate costs and effects. Clin Microbiol Infect 16:1754–1761 [View Article][PubMed]
WillemsenI., OverdevestI., Al NaiemiN., RijnsburgerM., SavelkoulP., Vandenbroucke-GraulsC., KluytmansJ.TRIANGLe Study Group2011; New diagnostic microarray (Check-KPC ESBL) for detection and identification of extended-spectrum β-lactamases in highly resistant Enterobacteriaceae. J Clin Microbiol 49:2985–2987 [View Article][PubMed]
WuT.-L., SiuL. K., SuL.-H., LauderdaleT. L., LinF. M., LeuH.-S., LinT.-Y., HoM.2001; Outer membrane protein change combined with co-existing TEM-1 and SHV-1 β-lactamases lead to false identification of ESBL-producing Klebsiella pneumoniae. J Antimicrob Chemother 47:755–761 [View Article][PubMed]