耐药克雷伯菌BlaNDM基因的表型及分子检测。

Taqdees Malik, Asma Naim
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引用次数: 0

摘要

背景:在过去的几个世纪里,克雷伯氏菌对抗菌素的耐药性普遍增加。克雷伯氏菌属的抗生素耐药菌株正成为临床环境中的严重威胁,因为它们涉及严重的侵入性和非侵入性感染。在这些菌株中出现耐药性与它们对几种广谱抗生素的强酶活性有关。这些酶包括β -内酰胺酶、扩展谱β -内酰胺酶(ESBL)、AmpC β -内酰胺酶和碳青霉烯酶。这些耐药酶能够水解各种广谱药物,如广谱头孢菌素和碳青霉烯类药物。目的:本研究通过聚合酶链反应鉴定克雷伯菌碳青霉烯酶的表型产生和NDM耐药基因的频率,以确定克雷伯菌中出现的耐药性。方法:本研究对来自不同临床实验室的236株革兰氏阴性分离株进行了鉴定。用标准微生物学方法鉴定出125株克雷伯氏菌。采用宏肉汤稀释法测定8种代表性抗生素的最低抑菌浓度(MIC)。采用改良霍奇法对产碳青霉烯酶克雷伯菌进行表型检测。然后对表型结果进行检查,并与利用聚合酶链反应(PCR)检测碳青霉烯酶抗性基因获得的基因型结果进行比较。结果:在本研究中,通过表型方法,仅在6株(5%)克雷伯菌分离株中发现碳青霉烯酶产生;然而,125株分离株中有3株NDM-1基因阳性。结论:由于我们考虑将碳青霉烯类药物作为治疗主要由革兰氏阴性分离株引起的感染的最后治疗选择,因此对该药物的普遍耐药正在广泛传播。为了在卫生保健机构、医院、实验室等实施严格的抗生素控制政策,最好评估抗生素敏感性,进行表型筛查和基因型筛查(如果可能)。
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Phenotypic and Molecular Detection of BlaNDM Gene Among Drug-Resistant Klebsiella Isolates.

Background: In the past few centuries, a widespread increase in antimicrobial resistance has been observed among Klebsiella species. The antibiotic- resistant strains of the genus Klebsiella are becoming a serious threat in clinical settings due to their involvement in severe invasive and non-invasive infections. The emergence of resistance among these strains is associated with their strong enzymatic activity against several broad-spectrum antibiotics. These enzymes include beta-lactamases, extended-spectrum beta-lactamases (ESBL), AmpC beta-lactamases, and carbapenemases. These resistance enzymes are capable of hydrolyzing various broad-spectrum drugs like extended-spectrum cephalosporin and carbapenems.

Objective: The present study was conducted to determine the emerging resistance among Klebsiella strains by identifying the production of carbapenemase enzyme phenotypically and the frequency of the NDM resistance gene by a polymerase chain reaction.

Methods: In this study, 236 Gram-negative isolates from different clinical laboratories were identified. Out of which, 125 isolates were found as Klebsiella species by using standard microbiological techniques. Minimum inhibitory concentrations (MIC) were determined using eight representative antibiotics by the Macro broth dilution method. Phenotypic detection of carbapenemase producing Klebsiella species was performed by Modified Hodge Test. Phenotypic findings were then checked and compared with genotypic results obtained by using the Polymerase chain reaction (PCR) for the detection of resistance genes responsible for the production of carbapenemase.

Results: In this study, carbapenemase production was found only in 6 (5%) Klebsiella isolates by using the phenotypic method; however, 3 isolates out of 125 were screened positive for the gene NDM-1.

Conclusion: Since we are considering carbapenems as the last therapeutic option for treating infections, mainly caused by Gram-negative isolates, the prevailing resistance against this drug is widely disseminating. It is better to evaluate the antibiotic susceptibility, phenotypic screening as well genotypic screening (where possible) for implementing strict antibiotic control policies in health care settings, hospitals, laboratories, etc.

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