检测来自巴基斯坦白沙瓦人类临床样本中多重耐药的大肠杆菌和肺炎克雷伯菌的 MCR-1 基因。
Detection of MCR-1 Gene in Multiple Drug Resistant Escherichia coli and Klebsiella pneumoniae in Human Clinical Samples from Peshawar, Pakistan.
机构信息
Institute of Basic Medical Sciences, Khyber Medical University, Peshawar, Pakistan.
Department of Physical Sciences and Information Technology, Faculty of Health Sciences, Anhui University, Hefei, China.
出版信息
Comb Chem High Throughput Screen. 2021;24(5):737-742. doi: 10.2174/1386207323666200914100119.
BACKGROUND
The presence of plasmid mediated mcr-1 gene in multidrug resistant Gram-negative bacteria poses a serious public health concern in today's world.
OBJECTIVE
The present study was aimed to detect the presence of plasmid mediated mcr-1 encoding resistance to colistin in multiple drug resistant (MDR) E. coli and K. pneumoniae isolates.
METHODS
A total of 180 clinical isolates of E. coli (n=120) and K. pneumoniae (n=60) were isolated from different clinical specimens, i.e., urine, blood, stool and pus, from diagnostic labs of two major public sector tertiary care hospitals in Peshawar, Pakistan. MDR profile of these isolates was assessed through Kirby-Baur disc diffusion method. All isolates were screened for colistin resistance by dilution methods. Colistin resistant isolates were subjected to PCR for mcr-1 detection and confirmation was done by Sanger sequencing method.
RESULTS
Overall, 83.3% (100/120) E. coli and 93.3% (56/60) K. pneumoniae were detected as MDR. Colistin resistance was found in 23.3% (28/120) E. coli and 40% (24/60) K. pneumoniae isolates, whereas mcr-1 gene was detected in 10 out of 52 colistin resistant isolates, including six E. coli and four K. pneumoniae isolates. Minimum inhibitory concentrations (MICs) of colistin in these ten mcr-1 positive isolates ranged from 4μg/ml to 16μg/ml. All mcr-1 positive isolates showed 99% sequence similarity when compared with other present sequences in GenBank.
CONCLUSION
Hence, our study confirms the presence of mcr-1 mediated colistin resistance in the studied area. Therefore, urgently larger scale surveillance studies are recommended to investigate prevalence of mcr-1 mediated colistin resistance and to prevent its further spread in the area.
背景
在当今世界，多药耐药革兰氏阴性菌中存在质粒介导的 mcr-1 基因，这对公共卫生构成了严重威胁。
目的
本研究旨在检测多药耐药（MDR）大肠杆菌和肺炎克雷伯菌分离株中质粒介导的 mcr-1 编码对多粘菌素的耐药性。
方法
从巴基斯坦白沙瓦两家主要公立医院的诊断实验室收集来自不同临床标本（尿液、血液、粪便和脓液）的 180 株临床分离株，包括 120 株大肠杆菌和 60 株肺炎克雷伯菌。通过 Kirby-Baur 圆盘扩散法评估这些分离株的 MDR 谱。所有分离株均通过稀释法筛查对多粘菌素的耐药性。对多粘菌素耐药的分离株进行 mcr-1 检测的 PCR，并通过 Sanger 测序法进行确认。
结果
总体而言，120 株大肠杆菌中有 83.3%（100/120）和 60 株肺炎克雷伯菌中有 93.3%（56/60）被检测为 MDR。在 28/120 株大肠杆菌和 24/60 株肺炎克雷伯菌分离株中发现对多粘菌素的耐药性，而在 52 株多粘菌素耐药分离株中检测到 10 株 mcr-1 基因，包括 6 株大肠杆菌和 4 株肺炎克雷伯菌分离株。这 10 株 mcr-1 阳性分离株的多粘菌素最小抑菌浓度（MIC）范围为 4μg/ml 至 16μg/ml。当与 GenBank 中的其他现有序列进行比较时，所有 mcr-1 阳性分离株的序列相似度均为 99%。
结论
因此，我们的研究证实了在所研究地区存在 mcr-1 介导的多粘菌素耐药性。因此，建议紧急进行更大规模的监测研究，以调查 mcr-1 介导的多粘菌素耐药性的流行情况，并防止其在该地区进一步传播。
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