| Autor: |
Hindiyeh, Musa, Smollen, Gill, Grossman, Zehava, Ram, Daniela, Davidson, Yehudit, Mileguir, Fernando, Vax, Marina, Ben David, Debbie, Tal, Ilana, Rahav, Galia, Shamiss, Ari, Mendelson, Ella, Keller, Nathan |
| Zdroj: |
Journal of Clinical Microbiology; September 2008, Vol. 46 Issue: 9 p2879-2883, 5p |
| Abstrakt: |
ABSTRACTCarbapenem resistance among Enterobacteriaceaeis an emerging problem worldwide. Klebsiella pneumoniaecarbapenemase (blaKPC) enzymes are among the most common ß-lactamases described. In this study, we report the development and validation of a real-time PCR (q-PCR) assay for the detection of blaKPCgenes using TaqMan chemistry. The q-PCR amplification of blaKPCDNA was linear over 7 log dilutions (r2= 0.999; slope, 3.54), and the amplification efficiency was 91.6%. The q-PCR detection limit was 1 CFU, and there was no cross-reaction with DNA extracted from several multidrug-resistant bacteria. Perianal/rectal swabs (n= 187) collected in duplicate from 128 patients admitted to Sheba Medical Center surgical intensive care units were evaluated for the presence of carbapenem-resistant bacteria by culturing on MacConkey agar-plus-carbapenem disks and for blaKPCgenes by q-PCR. Carbapenem-resistant organisms, all K. pneumoniae, were isolated from 47 (25.1%) of the 187 samples collected, while blaKPCgenes were detected in 54 (28.9%) of the patient samples extracted by the NucliSENS easyMAG system. Of these, seven samples were positive for blaKPCgenes by q-PCR but negative for carbapenem resistance by culture, while all samples in which no carbapenem-resistant bacteria were detected by culture also tested negative by q-PCR. Thus, the sensitivity and specificity of the q-PCR assay after extraction by the NucliSENS easyMAG system were 100% and 95%, respectively. Similar values were obtained after DNA extraction by the Roche MagNA Pure LC instrument: 97.9% sensitivity and 96.4% specificity. Overall, the blaKPCq-PCR assay appears to be highly sensitive and specific. The utilization of q-PCR will shorten the time to blaKPCdetection from 24 h to 4 h and will help in rapidly isolating colonized or infected patients and assigning them to cohorts. |
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