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三级护理医院临床样本分离的凝固酶阴性葡萄球菌中mecA基因的检测及其与抗菌药物耐药性的关系:一项横断面研究

Detection of the mecA Gene and Its Association With Antimicrobial Resistance Among Coagulase-Negative Staphylococci Isolated From Clinical Samples in a Tertiary Care Hospital: A Cross-Sectional Study.

作者信息

Tluanpuii Vanlal, Mahajan Rakesh K

机构信息

Microbiology, Atal Bihari Vajpayee Institute of Medical Sciences and Dr. Ram Manohar Lohia Hospital, New Delhi, IND.

出版信息

Cureus. 2025 Apr 3;17(4):e81643. doi: 10.7759/cureus.81643. eCollection 2025 Apr.

Abstract

Introduction Coagulase-Negative Staphylococci (CoNS) have become major pathogens causing a variety of infections. Antimicrobial resistance, brought about by the acquisition of the A gene, further exacerbates this burden. This gene encodes the altered membrane-bound protein called penicillin-binding protein (PBP2a), which lowers the binding affinity for β-lactam antibiotics. The widespread distribution of this gene has led to the surge of methicillin-resistant CoNS (MR-CoNS). Therefore, this study aimed to detect the prevalence of the A gene and its correlation with antimicrobial resistance. Methods This was a cross-sectional observational study conducted over a period from January 2021 to May 2022 at a tertiary care hospital in Delhi, India. A total of 100 pure isolates of CoNS from various samples were included and identified at the species level using conventional methods and an automated Vitek-2 ID system (bioMérieux, Marcy-l'Étoile, France). Antibiotic susceptibility testing was performed using the Kirby-Bauer disk diffusion method, as per the Clinical and Laboratory Standards Institute (CLSI) 2020 guideline, with the following antibiotics: cefoxitin (30 μg), erythromycin (15 μg), cotrimoxazole (1.25/23.75 μg), tetracycline (30 μg), gentamicin (10 μg), and ciprofloxacin (5 μg). The minimum inhibitory concentration (MIC) for vancomycin was detected using the Epsilometer test (E-test). All of the 100 isolates were also subjected to real-time polymerase chain reaction (rt-PCR) for the detection of the A gene. Results Out of 100 CoNS isolates, was the most commonly isolated, accounting for 44%. Cefoxitin was sensitive for only 20% of the isolates. Sensitivities for ciprofloxacin, cotrimoxazole, gentamicin, tetracycline, and erythromycin were 56%, 55%, 50%, 49%, and 25%, respectively. All isolates were sensitive to vancomycin. Out of 100 CoNS isolates that were subjected to A gene detection by rt-PCR, 82 (82%) isolates were positive for the A gene, and the rest were negative. Among the 82 (82%) A gene-positive isolates, seven (8.5%) were found to be cefoxitin sensitive, and among the 18 (18%) that were negative for the A gene, cefoxitin resistance was detected in five (27.7%) isolates by disk diffusion method. Both cefoxitin disk diffusion and A gene detection by rt-PCR were employed to detect methicillin resistance among the CoNS isolates. Conclusion The A gene was detected in 82 (82%) isolates. However, some A-positive CoNS were found to be susceptible to cefoxitin by the disk diffusion method, while some A-negative CoNS isolates were resistant. This suggests that no single diagnostic approach is capable of identifying methicillin resistance alone. Therefore, A gene detection must be implemented along with cefoxitin disk diffusion to make accurate and early therapeutic decisions.

摘要

引言 凝固酶阴性葡萄球菌(CoNS)已成为引起各种感染的主要病原体。通过获得A基因产生的抗菌药物耐药性进一步加重了这一负担。该基因编码一种称为青霉素结合蛋白(PBP2a)的改变的膜结合蛋白,它降低了对β-内酰胺类抗生素的结合亲和力。该基因的广泛分布导致了耐甲氧西林CoNS(MR-CoNS)的激增。因此,本研究旨在检测A基因的流行情况及其与抗菌药物耐药性的相关性。

方法 这是一项横断面观察性研究,于2021年1月至2022年5月在印度德里的一家三级护理医院进行。共纳入100株来自各种样本的CoNS纯分离株,并使用传统方法和自动化Vitek-2 ID系统(法国生物梅里埃公司,马西-埃图瓦勒)在种水平上进行鉴定。根据临床和实验室标准协会(CLSI)2020年指南,使用 Kirby-Bauer 纸片扩散法对以下抗生素进行药敏试验:头孢西丁(30 μg)、红霉素(15 μg)、复方新诺明(1.25/23.75 μg)、四环素(30 μg)、庆大霉素(10 μg)和环丙沙星(5 μg)。使用Epsilometer试验(E试验)检测万古霉素的最低抑菌浓度(MIC)。所有100株分离株均进行实时聚合酶链反应(rt-PCR)以检测A基因。

结果 在100株CoNS分离株中, 是最常分离到的,占44%。头孢西丁仅对20%的分离株敏感。环丙沙星、复方新诺明、庆大霉素、四环素和红霉素的敏感性分别为56%、55%、50%、49%和25%。所有分离株对万古霉素敏感。在通过rt-PCR进行A基因检测的100株CoNS分离株中,82株(82%)A基因检测呈阳性,其余为阴性。在82株(82%)A基因阳性分离株中,7株(8.5%)被发现对头孢西丁敏感,在18株(18%)A基因阴性分离株中,通过纸片扩散法检测到5株(27.7%)分离株对头孢西丁耐药。同时采用头孢西丁纸片扩散法和rt-PCR检测A基因来检测CoNS分离株中的耐甲氧西林情况。

结论 在82株(82%)分离株中检测到A基因。然而,通过纸片扩散法发现一些A阳性CoNS对头孢西丁敏感,而一些A阴性CoNS分离株耐药。这表明没有单一的诊断方法能够单独识别耐甲氧西林情况。因此,必须同时进行A基因检测和头孢西丁纸片扩散法,以做出准确和早期的治疗决策。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee43/12048871/1421461eb87e/cureus-0017-00000081643-i01.jpg

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