Department of Microbiology, Assam University, Silchar, India.
Department of Microbiology, Silchar Medical College & Hospital, Silchar, India.
Indian J Med Res. 2017 May;145(5):659-664. doi: 10.4103/ijmr.IJMR_628_15.
BACKGROUND & OBJECTIVES: Pseudomonas aeruginosa possessing chromosomally inducible blaPDCalong with other intrinsic mechanism causes infection with high mortality rate. It is difficult to detect inducible AmpC enzymes in this organism and is usually overlooked by routine testing that may lead to therapeutic failure. Therefore, three different inducers were evaluated in the present study to assess their ability of induction of blaPDCin P. aeruginosa.
A total of 189 consecutive Pseudomonas isolates recovered from different clinical specimens (November 2011-April 2013) were selected for the study. Isolates were screened with cefoxitin for AmpC β-lactamases and confirmed by modified three-dimensional extract test (M3DET). Inductions were checked using three inducers, namely, clavulanic acid, cefoxitin and imipenem along with ceftazidime. Molecular screening of AmpC β-lactamase genes was performed by PCR assay. Antimicrobial susceptibility and minimum inhibitory concentrations (MICs) were determined, and repetitive extragenic palindromic-PCR of all blaPDCharbouring isolates was performed.
Inducible phenotype was observed in 42 (24.3%) of 97 (56%) isolates confirmed by M3DET. Among these, 22 isolates harboured chromosomal blaPDCgene, and cocarriage of both chromosomal and plasmid-mediated blaAmpC genes was observed in seven isolates. Cefoxitin-ceftazidime-based test gave good sensitivity and specificity for detecting inducible AmpC enzymes. Isolates harbouring blaPDCshowed high MIC against all tested cephalosporins and monobactam. DNA fingerprinting of these isolates showed 22 different clones of P. aeruginosa.
INTERPRETATION & CONCLUSIONS: P. aeruginosa harbouring inducible (chromosomal) and plasmid-mediated AmpC β-lactamase is a matter of concern as it may limit therapeutic option. Using cefoxitin-ceftazidime-based test is simple and may be used for detecting inducible AmpC β-lactamase amongst P. aeruginosa.
铜绿假单胞菌具有可诱导的 blaPDC 染色体基因,同时还具有其他内在机制,导致感染死亡率高。在这种生物体中检测诱导型 AmpC 酶很困难,并且通常会被常规检测所忽略,这可能导致治疗失败。因此,本研究评估了三种不同的诱导剂,以评估它们在诱导铜绿假单胞菌 blaPDC 中的能力。
选择了 189 株连续分离自不同临床标本的铜绿假单胞菌(2011 年 11 月至 2013 年 4 月)进行研究。用头孢西丁筛选 AmpC β-内酰胺酶,并通过改良的三维提取试验(M3DET)确认。使用三种诱导剂,即克拉维酸、头孢西丁和亚胺培南以及头孢他啶来检查诱导作用。采用 PCR 法进行 AmpC β-内酰胺酶基因的分子筛选。测定抗菌药物敏感性和最低抑菌浓度(MIC),并对所有 blaPDC 携带菌株进行重复外回文 PCR。
通过 M3DET 确认的 97 株(56%)中有 42 株(24.3%)表现出诱导表型。其中,22 株携带染色体 blaPDC 基因,7 株同时携带染色体和质粒介导的 blaAmpC 基因。头孢西丁-头孢他啶试验对检测诱导型 AmpC 酶具有良好的敏感性和特异性。携带 blaPDC 的菌株对所有测试的头孢菌素和单酰胺类药物均显示出较高的 MIC。这些菌株的 DNA 指纹图谱显示了 22 种不同的铜绿假单胞菌克隆。
携带可诱导(染色体)和质粒介导的 AmpC β-内酰胺酶的铜绿假单胞菌是一个值得关注的问题,因为它可能会限制治疗选择。使用头孢西丁-头孢他啶试验是一种简单的方法,可用于检测铜绿假单胞菌中的诱导型 AmpC β-内酰胺酶。