Peptide Spectra Analysis of DNA Starvation/Stationary Phase Protection-Like Protein Found in Hospital-Acquired ESBL Producing Escherichia coli by MALDI-TOF Technique

Purilarp Daoaroonkiet, Siriaran Kwangsukstith, Salin Srichai, Swasamon Jaidee, Metha Emthanom, Pijitra Sujitpiriyakit, T. Sastraruji, S. Sookkhee
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Abstract

The present study aimed to investigate the hospital-acquired Extended Spectrum β-Lactamase production (ESBL) producing Escherichia coli, to identify the significant ESBL peptide spectra after exposure to ceftazidime and amoxicillin/clavulanic acid, and to identify the resistant peptide spectra after exposure to ciprofloxacin. ESBL production and the minimal inhibitory concentration of ciprofloxacin were investigated using the Epsilometer strip test. Their colonies were collected to analyze the peptide spectra by VITEK® Mass Spectrometry. The significant difference in spectrum intensity for suspected ESBL peptide spectra, and DNA starvation/stationary phase protection (Dps) -like peptide spectra were also determined. Among 228 isolates, the high ESBL-producing E. coli which exhibited high ciprofloxacin resistance was demonstrated as 22.63%. The significant reduction of spectrum intensity after exposure to ceftazidime and amoxicillin/clavulanic acid was only detected at spectrum 9714 Da in the high ESBL-producing group (P ≤0.001). Among four selected high ESBL producers, only one significant spectrum located at 9063 Da was detected in the high ciprofloxacin-resistant group (P ≤0.001) after being exposed to ceftazidime and ciprofloxacin. The highest percentage increase of spectrum 9063 Da was 69.42%. Multidrug resistance was demonstrated in the tested E. coli isolates, especially β-lactam resistance which may be caused by their ESBL production. It may be said that the peptide spectrum located at 9714, and 9063 Da were suspected as the ESBL indicative peptide spectrum in the high ESBL producers, and as the Dps indicative peptide spectrum in the high ciprofloxacin-resistant isolates. Keywords: Peptide spectra analysis, DNA starvation/stationary phase protection like protein, Hospital-acquired infection, Extended spectrum β-lactamase, Escherichia coli, Matrix-Assisted Laser Desorption Ionization-Time Of Flight
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MALDI-TOF技术分析医院获得性ESBL大肠杆菌中DNA饥饿/固定相保护样蛋白的肽谱
本研究旨在研究医院获得性β-内酰胺酶生产(ESBL)大肠杆菌,确定暴露于头孢他啶和阿莫西林/克拉维酸后的显著ESBL肽谱,以及暴露于环丙沙星后的耐药肽谱。采用Epsilometer试纸法考察了ESBL的产生和环丙沙星的最低抑菌浓度。收集其菌落,用VITEK®质谱法分析肽谱。可疑ESBL肽谱和DNA饥饿/固定相保护(Dps)样肽谱的光谱强度也有显著差异。228株分离株中,产esbl高的大肠杆菌占22.63%,对环丙沙星有较高的耐药性。暴露于头孢他啶和阿莫西林/克拉维酸后,仅在esbl高生成组的光谱9714da处检测到光谱强度的显著降低(P≤0.001)。在所选的4个ESBL高耐药菌株中,暴露于头孢他啶和环丙沙星的高耐药组仅检测到一个位于9063 Da的显著谱(P≤0.001)。光谱9063 Da增幅最大,达69.42%。在大肠杆菌分离株中发现了多药耐药,特别是β-内酰胺耐药,这可能是由它们产生ESBL引起的。可以推测,位于9714和9063 Da的肽谱可能是高ESBL生产者的ESBL指示肽谱,而位于高环丙沙星耐药菌株的Dps指示肽谱。关键词:肽谱分析,DNA饥饿/固定相保护如蛋白质,医院获得性感染,扩展谱β-内酰胺酶,大肠杆菌,基质辅助激光解吸电离飞行时间
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