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[生物膜产生菌和非生物膜产生菌金黄色葡萄球菌临床分离株的疏水特性研究]

[Investigation of hydrophobic characteristics of biofilm producer and non-producer Staphylococcus aureus clinical isolates].

作者信息

Ay Selma, Güldür Tayfun, Tekerekoğlu Mehmet Sait, Otlu Baniş

机构信息

Inönü Universitesi Tip Fakültesi, Tibbi Mikrobiyoloji Anabilim Dali, Malatya.

出版信息

Mikrobiyol Bul. 2010 Apr;44(2):221-30.

Abstract

The ability of staphylococcus to adhere certain structures and to form biofilm (slime) layer plays an important role in the pathogenesis of staphylococcal infections. Hydrophobic interactions and hydrogen bonds are important factors that play role in adherence. This study was designed to compare the hydrophobic properties of slime positive and negative Staphylococcus aureus strains isolated from blood cultures. Ten methicillin-resistant S. aureus isolates (five of them being slime positive) obtained from blood cultures of patients at intensive care unit of a university hospital, between May 2006 and June 2007, were included in the study. Slime production of the isolates was determined by Christensen's method. Methicillin resistance was determined by cefoxitin disc test and oxacillin salt agar test. It was determined that the test strains did not exhibit any autoaggregation. The adherence of strains to the three different hydrocarbons as solid phases (butyl-sepharose, octyl-sepharose and phenyl-sepharose; Amersham Bioscience, Sweden) were studied by using hydrophobic interaction chromatography (HIC) method. After butyl- and octyl-sepharose chromatography, it was determined that slime negative S. aureus strains were separated into three fractions eluted with phosphate buffered saline (PBS), 40% and 96% ethanol, while slime positive strains were separated into two fractions eluted with 40% and 96% ethanol, respectively. By phenyl-sepharose chromatography analysis; both slime negative and positive strains were separated into two fractions eluted in 40% and 96% ethanol. Hydrophobicity tests were repeated at 4 degrees C and pH 6-9 to evaluate the effect of changing conditions on hydrophobicity. However, no changes we re observed at these temperature and pH values. According to these analysis it was concluded that; (a) S. aureus strains consist heterogeneous fractions with distinct hydrophobic binding strengths; (b) hydrophobic surface protein secretion may be different in heterogeneous groups, and (c) slime positive S. aureus strains were more hydrophobic than non-slime producing strains. Further research is required in order to characterise the eluted fractions and to evaluate their pathogenic capacities.

摘要

葡萄球菌黏附某些结构并形成生物膜(黏液)层的能力在葡萄球菌感染的发病机制中起着重要作用。疏水相互作用和氢键是参与黏附的重要因素。本研究旨在比较从血培养中分离出的黏液阳性和阴性金黄色葡萄球菌菌株的疏水特性。研究纳入了2006年5月至2007年6月期间从一所大学医院重症监护病房患者的血培养中获得的10株耐甲氧西林金黄色葡萄球菌分离株(其中5株为黏液阳性)。采用克里斯滕森方法测定分离株的黏液产生情况。通过头孢西丁纸片试验和苯唑西林盐琼脂试验测定耐甲氧西林情况。结果表明测试菌株未表现出任何自聚集现象。采用疏水相互作用色谱(HIC)法研究菌株对三种不同的作为固定相的碳氢化合物(丁基琼脂糖、辛基琼脂糖和苯基琼脂糖;瑞典阿默沙姆生物科学公司)的黏附情况。经过丁基和辛基琼脂糖色谱分析后,发现黏液阴性金黄色葡萄球菌菌株被分离成用磷酸盐缓冲盐水(PBS)、40%和96%乙醇洗脱的三个组分,而黏液阳性菌株分别被分离成用40%和96%乙醇洗脱的两个组分。通过苯基琼脂糖色谱分析,黏液阴性和阳性菌株均被分离成用40%和96%乙醇洗脱的两个组分。在4℃和pH 6 - 9条件下重复进行疏水性测试,以评估条件变化对疏水性的影响。然而,在这些温度和pH值下未观察到变化。根据这些分析得出以下结论:(a)金黄色葡萄球菌菌株由具有不同疏水结合强度的异质组分组成;(b)疏水表面蛋白分泌在异质组中可能不同;(c)黏液阳性金黄色葡萄球菌菌株比不产生黏液的菌株疏水性更强。为了表征洗脱组分并评估其致病能力,还需要进一步研究。

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