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比较 MAMA PCR 和 SSCP PCR 以研究大肠杆菌和肺炎克雷伯菌对环丙沙星和萘啶酸的染色体耐药性。

The comparison of MAMA PCR and SSCP PCR to study chromosomal resistance against Ciprofloxacin and Nalidixic acid in Escherichia coli and Klebsiella pneumoniae.

机构信息

Department of Microbiology, Student Research Committee, Mazandaran University of Medical Sciences, Sari, Iran.

Department of Immunology, Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran.

出版信息

Microb Pathog. 2018 Jul;120:181-186. doi: 10.1016/j.micpath.2018.05.005. Epub 2018 May 6.

Abstract

The mutation in gyrA and parC genes alters amino acids. Also, it causes resistance against Fluoroquinolones in E. coli and K. pneumoniae. The purpose of this study was to diagnose the significant mutation of gyrA (ser83-asp87) and parC (ser80-glu84) genes through using MAMA PCR and SSCP PCR methods. In so doing, the isolated samples were collected. Then, utilizing agar disc diffusion method, the researchers performed antibiotic sensitivity test. Moreover, Fluoroquinolones resistance was confirmed by E-test method (MIC experiment). Furthermore, the obtained data from MAMA PCR method were sequenced accidentally. According to the findings, among 103 isolated samples, 65 samples (63/2%) were belonged to E. coli and 38 samples (36/8%) to K. pneumoniae. In all E. coli that resisted to Ciprofloxacin, at least one mutation were observed. Also, at least one mutation was observed in all K. pneumoniae samples that resisted to Ciprofloxacin. However, four mutation points were detected for each of seven samples and, interestingly, there was no mutation in five sensitive samples to Ciprofloxacin. In addition, the results revealed that the mutation in gyrA and parC genes was closely related to Quinolones resistance. Based on the findings, preparing an infection control program in Iran is highly required.

摘要

gyrA 和 parC 基因突变改变了氨基酸。此外,它导致大肠杆菌和肺炎克雷伯菌对氟喹诺酮类药物产生耐药性。本研究的目的是通过使用 MAMA PCR 和 SSCP PCR 方法诊断 gyrA(ser83-asp87)和 parC(ser80-glu84)基因的重要突变。为此,收集了分离的样本。然后,研究人员利用琼脂扩散法进行抗生素敏感性试验。此外,通过 E 试验法(MIC 实验)确认氟喹诺酮类药物的耐药性。此外,MAMA PCR 方法获得的数据意外地进行了测序。研究结果显示,在 103 个分离样本中,有 65 个(63/2%)属于大肠杆菌,38 个(36/8%)属于肺炎克雷伯菌。所有对环丙沙星耐药的大肠杆菌至少观察到一个突变。同样,所有对环丙沙星耐药的肺炎克雷伯菌样本中至少观察到一个突变。然而,有趣的是,在对环丙沙星敏感的五个样本中没有发现突变,七个样本中的每个样本都检测到了四个突变点。此外,结果表明 gyrA 和 parC 基因突变与喹诺酮类药物耐药性密切相关。基于这些发现,在伊朗制定感染控制计划是非常必要的。

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