Yıldız Serap Süzük, Kaşkatepe Banu, Avcıküçük Havva, Öztürk Şükran
1 National Antimicrobial Resistance Laboratory, Public Health Institution of Turkey , Ankara, Turkey.
2 Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Ankara University , Ankara, Turkey.
Acta Microbiol Immunol Hung. 2017 Mar 1;64(1):9-16. doi: 10.1556/030.64.2017.009.
This study applied two phenotypic tests, namely "Carbapenemase Nordmann-Poirel" (CarbaNP) test and "Carbapenem Inactivation Method" (CIM), against the isolates carrying the carbapenem resistance genes. The study included 83 carbapenem-resistant Enterobacteriaceae isolates producing oxacillinase-48 (OXA-48) and 30 carbapenem-sensitive Enterobacteriaceae isolates. Out of the total isolates studied, 77 isolates (92.77%) were identified as Klebsiella pneumoniae and six isolates (7.23%) were identified as Escherichia coli by Matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry. Polymerase chain reaction (PCR) method used to detect resistance genes found that 74 isolates (89.16%) produced OXA-48 carbapenemase, whereas nine isolates (10.84%) produced both OXA-48 and New Delhi metallo-beta-lactamase-1 (NDM-1). The isolates producing both OXA-48 and NDM-1 were found to be positive by both phenotypic tests. Among isolates carrying only bla gene alone, nine isolates (13.04%) for CarbaNP test and two isolates for CIM test (2.90%) displayed false negative results, respectively. The sensitivity of CarbaNP and CIM tests was found to be 89.16% and 97.59%, respectively, whereas the specificity was determined to be 100% for both tests. These findings suggest that CarbaNP and CIM tests are useful tools to identify the carbapenemase producers. Molecular methods like PCR are recommended to verify false negative tests predicted to have OXA-48 activity.
本研究对携带碳青霉烯类耐药基因的分离株应用了两种表型检测方法,即“碳青霉烯酶诺德曼 - 普雷尔”(CarbaNP)检测和“碳青霉烯灭活法”(CIM)。该研究纳入了83株产氧亚胺酶 - 48(OXA - 48)的耐碳青霉烯类肠杆菌科分离株和30株对碳青霉烯类敏感的肠杆菌科分离株。在所研究的全部分离株中,通过基质辅助激光解吸电离飞行时间质谱法鉴定出77株(92.77%)为肺炎克雷伯菌,6株(7.23%)为大肠埃希菌。用于检测耐药基因的聚合酶链反应(PCR)方法发现,74株(89.16%)产生OXA - 48碳青霉烯酶,而9株(10.84%)同时产生OXA - 48和新德里金属β - 内酰胺酶 - 1(NDM - 1)。同时产生OXA - 48和NDM - 1的分离株在两种表型检测中均呈阳性。在仅携带bla基因的分离株中,CarbaNP检测有9株(13.04%)、CIM检测有2株(2.90%)分别出现假阴性结果。发现CarbaNP检测和CIM检测的敏感性分别为89.16%和97.59%,而两种检测的特异性均为100%。这些发现表明,CarbaNP检测和CIM检测是鉴定碳青霉烯酶产生菌的有用工具。建议采用PCR等分子方法来验证预计具有OXA - 48活性的假阴性检测结果。