Pruss Agata, Skierska Alicja, Kwiatkowski Paweł, Masiuk Helena, Jursa-Kulesza Joanna, Giedrys-Kalemba Stefania, Bilska Iwona, Sienkiewicz Monika, Melnyk Maria V, Dołęgowska Barbara
Department of Laboratory Medicine, Chair of Microbiology, Immunology and Laboratory Medicine, Pomeranian Medical University in Szczecin, Szczecin, Poland.
Department of Diagnostic Immunology, Chair of Microbiology, Immunology and Laboratory Medicine, Pomeranian Medical University in Szczecin, Szczecin, Poland.
PLoS One. 2025 Feb 7;20(2):e0318852. doi: 10.1371/journal.pone.0318852. eCollection 2025.
Klebsiella pneumoniae is a typical opportunistic pathogen that exhibits multiple virulence factors and antibiotic resistance conditioning mechanisms. Carbapenemases are enzymes that help bacteria to exhibit the strongest resistance against antibiotics. Therefore, in routine microbiological diagnoses, it is crucial to confirm antibiotic-resistant strains, including carbapenemase-producing bacteria strains, isolated from patients. Two types of tests play an important role here: phenotypicand molecular diagnostic methods. The latter complement phenotypic tests and a mandatory procedure to confirm the detection of carbapenemases. This study aimed to evaluate the usefulness and effectiveness of tests and methods used to identify and confirm the ability of clinical K. pneumoniae strains to produce carbapenemases. The production of carbapenemases was assessed using phenotypic and genetic methods. The strains tested showed complete resistance to most beta-lactams and varying sensitivity to drugs from the quinolone carbapenem group and aminoglycosides. Among the most commonly produced carbapenemases were the metallo-beta-lactamase (NDM) family. The most accurate phenotypic method for detecting carbapenemases was the NG CARBA-5 assay, and the PCR method confirmed these results. Notably, a few inconclusive results were obtained for NDM-positive and VIM-positive strains when the disk diffusion method and CIM test were used. Further, the Carba tube assay and the RAPIDEC CARBA NP assay produced questionable results for the OXA-48 strain group. This group also generated false-negative results on Carba's CHROM ID medium.
肺炎克雷伯菌是一种典型的机会致病菌,具有多种毒力因子和抗生素耐药调节机制。碳青霉烯酶是帮助细菌展现出最强抗生素耐药性的酶。因此,在常规微生物诊断中,确认从患者体内分离出的包括产碳青霉烯酶细菌菌株在内的耐药菌株至关重要。两种检测在此发挥重要作用:表型检测和分子诊断方法。后者补充表型检测,是确认碳青霉烯酶检测结果的强制性程序。本研究旨在评估用于鉴定和确认临床肺炎克雷伯菌菌株产生碳青霉烯酶能力的检测方法和手段的实用性和有效性。使用表型和基因方法评估碳青霉烯酶的产生情况。受试菌株对大多数β-内酰胺类药物表现出完全耐药,对喹诺酮碳青霉烯类药物和氨基糖苷类药物的敏感性各不相同。最常见产生的碳青霉烯酶属于金属β-内酰胺酶(NDM)家族。检测碳青霉烯酶最准确的表型方法是NG CARBA-5检测,PCR方法证实了这些结果。值得注意的是,当使用纸片扩散法和CIM检测时,NDM阳性和VIM阳性菌株获得了一些不确定的结果。此外,Carba tube检测和RAPIDEC CARBA NP检测对OXA-48菌株组产生了可疑结果。该菌株组在Carba的CHROM ID培养基上也产生了假阴性结果。