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加纳阿克拉产碳青霉烯酶大肠埃希菌和肺炎克雷伯菌的表型和基因型检测。

Phenotypic and genotypic detection of carbapenemase-producing Escherichia coli and Klebsiella pneumoniae in Accra, Ghana.

机构信息

Department of Medical Microbiology, University of Ghana Medical School, Korle Bu, Accra, Ghana.

Department of Medical Laboratory, University of Ghana Medical Centre, Legon, Accra, Ghana.

出版信息

PLoS One. 2022 Dec 30;17(12):e0279715. doi: 10.1371/journal.pone.0279715. eCollection 2022.

Abstract

AIM

To describe the occurrence of carbapenem resistance among multidrug-resistant (MDR) Escherichia coli and Klebsiella pneumoniae isolated from clinical specimens in Accra using phenotypic and genotypic methods.

METHODOLOGY

The study was cross-sectional, involving 144 clinical MDR E. coli and K. pneumoniae isolates recovered from the Central Laboratory of the Korle Bu Teaching Hospital (KBTH). The isolates were re-cultured bacteriologically, identified using standard biochemical tests, and subjected to antibiotic susceptibility testing using the Kirby-Bauer method. Carbapenem resistance was determined based on imipenem, meropenem, and ertapenem zones of inhibition, as well as minimum inhibitory concentrations (MICs). Carbapenemase production was determined phenotypically by modified Hodge test (MHT) and modified carbapenem inactivation method (mCIM), and genotypically with multiplex PCR targeting the blaKPC, blaIMP, blaNDM, blaVIM, and blaOXA-48 genes.

RESULTS

Of the 144 MDR isolates, 69.4% were E. coli, and 30.6% were K. pneumoniae. The distribution of antimicrobial resistance rates among them was ampicillin (97.2%), cefuroxime (93.1%), sulfamethoxazole-trimethoprim (86.8%), tetracycline (85.4%), cefotaxime and cefpodoxime (77.1% each), amoxicillin-clavulanate (75%), ceftriaxone (73.6%), ciprofloxacin (70.8%), levofloxacin (66.0%), cefepime (65.3%), ceftazidime (64.6%), gentamicin (48.6), piperacillin-tazobactam (40.3%), cefoxitin (14.6%), amikacin (13.9%), ertapenem and meropenem (5.6% each), and imipenem (2.8%). In total, 5.6% (8/144) of them were carbapenem-resistant (carbapenem MIC range = 0.094-32.0 μg/ml), with 75% (6/8) of these testing positive by the phenotypic tests and 62.5% (5/8) by the genotypic test (of which 80% [4/5] carried blaOXA-48 and 20% (1/5) blaNDM). The blaVIM, blaIMP, and blaKPC genes were not detected.

CONCLUSION

Although the rates of antibiotic resistance among the isolates were high, the prevalence of carbapenemase producers was low. The finding of blaOXA-48 and blaNDM warrants upscaling of antimicrobial resistance surveillance programmes and fortification of infection prevention and control programmes in the country.

摘要

目的

使用表型和基因型方法描述加纳阿克拉临床标本中分离的多药耐药(MDR)大肠埃希菌和肺炎克雷伯菌中产碳青霉烯酶的发生率。

方法

该研究为横断面研究,涉及从科勒布教学医院(KBTH)中心实验室回收的 144 株临床 MDR 大肠埃希菌和肺炎克雷伯菌。对分离物进行细菌培养再培养、使用标准生化试验进行鉴定,并使用 Kirby-Bauer 法进行抗生素敏感性试验。根据亚胺培南、美罗培南和厄他培南的抑制区以及最低抑菌浓度(MIC)来确定碳青霉烯类耐药性。通过改良 Hodge 试验(MHT)和改良碳青霉烯灭活法(mCIM)进行表型碳青霉烯酶产生检测,通过针对 blaKPC、blaIMP、blaNDM、blaVIM 和 blaOXA-48 基因的多重 PCR 进行基因型检测。

结果

在 144 株 MDR 分离株中,69.4%为大肠埃希菌,30.6%为肺炎克雷伯菌。它们的抗菌药物耐药率分布如下:氨苄西林(97.2%)、头孢呋辛(93.1%)、磺胺甲恶唑-甲氧苄啶(86.8%)、四环素(85.4%)、头孢噻肟和头孢泊肟(77.1%)、阿莫西林-克拉维酸(75%)、头孢曲松(73.6%)、环丙沙星(70.8%)、左氧氟沙星(66.0%)、头孢吡肟(65.3%)、头孢他啶(64.6%)、庆大霉素(48.6%)、哌拉西林-他唑巴坦(40.3%)、头孢西丁(14.6%)、阿米卡星(13.9%)、厄他培南和美罗培南(各 5.6%)和亚胺培南(2.8%)。总共,有 5.6%(8/144)的分离物为碳青霉烯类耐药(碳青霉烯类 MIC 范围=0.094-32.0 μg/ml),其中 75%(6/8)的分离物经表型试验呈阳性,62.5%(5/8)经基因型试验呈阳性(其中 80%(4/5)携带 blaOXA-48,20%(1/5)携带 blaNDM)。未检测到 blaVIM、blaIMP 和 blaKPC 基因。

结论

尽管分离物的抗生素耐药率较高,但产碳青霉烯酶的分离物比例较低。blaOXA-48 和 blaNDM 的发现需要扩大抗菌药物耐药性监测计划,并加强该国的感染预防和控制计划。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be2e/9803230/b9fdbe185ae3/pone.0279715.g001.jpg

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