Bae Il Kwon, Kang Hyun-Kyung, Jang In-Ho, Lee Woonhyoung, Kim Keonhan, Kim Jung Ok, Jeong Seok Hoon, Lee Kyungwon
Department of Dental Hygiene, Silla University, Busan, Korea.
Department of Biomedical Laboratory Science, College of Health Sciences, Sangji University, Wonju, Gangwon, Korea.
Infect Chemother. 2015 Sep;47(3):167-74. doi: 10.3947/ic.2015.47.3.167. Epub 2015 Sep 30.
The rapid and accurate detection of carbapenemase-producing Enterobacteriaceae (CPE) in clinical microbiology laboratories is essential for the treatment and control of infections caused by these microorganisms. This study was performed to evaluate the ability of the VITEK AST-N202 card to detect CPE isolates.
A total of 43 (Klebsiella pneumoniae, n = 37; Escherichia coli, n = 3; and Enterobacter cloacae, n = 3) CPE isolates and 79 carbapenemase-non-producing Enterobacteriaceae (CNE) isolates were included in this study. The CPE isolates harbored KPC-2 (n = 11), KPC-3 (n = 20), GES-5 (n = 5), VIM-2 (n = 2), IMP-1 (n = 1), NDM-1 (n = 2), or OXA-232 (n = 2). Of the 79 CNE isolates, eight K. pneumoniae isolates were resistant to ertapenem, imipenem, and meropenem, while the remaining 71 isolates were susceptible to the carbapenems. Antimicrobial susceptibilities were tested using the VITEK AST-N202 card, and the results were interpreted as positive when the isolates showed resistant or intermediate results. Modified-Hodge tests (MHTs) were performed using ertapenem or meropenem disks for the screening of carbapenemase production. Polymerase chain reaction (PCR) and direct sequencing were used to identify β-lactamase genes.
Sensitivity of MHT with ertapenem and meropenem disks for the detection of carbapenemase was 81.4% (35/43) and 81.4% (35/43), respectively, and a combination with both antibiotic disks increased the sensitivity to 88.4% (38/43). Specificity of the MHT was 100% (79/79) for the CNE isolates. Sensitivity of ertapenem, imipenem, and meropenem as assessed by the VITEK AST-N202 card was 100% (43/43), 93% (40/43), and 95.3% (41/43), respectively. Specificity (89.8%, 71/79) of the test with each carbapenem was improved to 100% (71/71) when eight carbapenem-resistant CNE isolates were excluded from the testing.
The VITEK AST-N202 card showed high sensitivity for the detection of carbapenemases in Enterobacteriaceae strains. PCR and sequencing experiments for the detection of carbapenemases are recommended when clinical Enterobacteriaceae isolates show non-susceptibility to carbapenems.
在临床微生物实验室中快速准确地检测产碳青霉烯酶肠杆菌科细菌(CPE)对于治疗和控制由这些微生物引起的感染至关重要。本研究旨在评估VITEK AST-N202卡片检测CPE分离株的能力。
本研究共纳入43株CPE分离株(肺炎克雷伯菌37株、大肠埃希菌3株、阴沟肠杆菌3株)和79株不产碳青霉烯酶肠杆菌科细菌(CNE)分离株。CPE分离株携带KPC-2(11株)、KPC-3(20株)、GES-5(5株)、VIM-2(2株)、IMP-1(1株)、NDM-1(2株)或OXA-232(2株)。在79株CNE分离株中,8株肺炎克雷伯菌分离株对厄他培南、亚胺培南和美罗培南耐药,其余71株对碳青霉烯类药物敏感。使用VITEK AST-N202卡片检测抗菌药物敏感性,当分离株显示耐药或中介结果时,结果判定为阳性。使用厄他培南或美罗培南纸片进行改良Hodge试验(MHT)以筛选碳青霉烯酶的产生。采用聚合酶链反应(PCR)和直接测序法鉴定β-内酰胺酶基因。
厄他培南纸片和美罗培南纸片MHT检测碳青霉烯酶的敏感性分别为81.4%(35/43)和81.4%(35/43),两种抗生素纸片联合使用可将敏感性提高至88.4%(38/43)。MHT对CNE分离株的特异性为100%(79/79)。VITEK AST-N202卡片评估的厄他培南、亚胺培南和美罗培南的敏感性分别为100%(43/43)、93%(40/43)和95.3%(41/43)。当从检测中排除8株耐碳青霉烯类CNE分离株时,每种碳青霉烯类药物检测的特异性(89.8%,71/79)提高到100%(71/71)。
VITEK AST-N202卡片对肠杆菌科菌株中碳青霉烯酶的检测具有较高的敏感性。当临床肠杆菌科分离株对碳青霉烯类药物不敏感时,建议进行PCR和测序实验检测碳青霉烯酶。