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脉冲场凝胶电泳作为皱落假丝酵母分型系统的评估:核型与限制性片段长度多态性的比较

Evaluation of pulsed-field gel electrophoresis as a typing system for Candida rugosa: comparison of karyotype and restriction fragment length polymorphisms.

作者信息

Dib J C, Dube M, Kelly C, Rinaldi M G, Patterson J E

机构信息

Department of Medicine/Infectious Diseases and Pathology, University of Texas Health Science Center at San Antonio 78284-7881, USA.

出版信息

J Clin Microbiol. 1996 Jun;34(6):1494-6. doi: 10.1128/jcm.34.6.1494-1496.1996.

Abstract

Nosocomial infections with Candida species have emerged as an increasingly important cause of morbidity and mortality in intensive care units. Ten Candida rugosa isolates from a previously documented cluster of C. rugosa infections in one hospital (nine burn unit isolates and one isolate from another hospital ward) and eight C. rugosa isolates recovered in a referral fungus testing laboratory (comparison isolates) from distinct geographic areas were investigated by molecular techniques. Isolates were from multiple anatomic sites. Pulsed-field gel electrophoresis (PFGE) of whole-cell DNA was performed with the 18 C. rugosa isolates as a marker of strain identity. The PFGE karyotypes of the C. rugosa isolates were demonstrated from four to seven chromosome bands. Karyotyping revealed the same PFGE pattern for the nine outbreak isolates from the burn unit, confirming clonal strain transmission. The isolate from the other hospital ward had a distinct karyotype. Distinct PFGE karyotype patterns were demonstrated for the eight comparison isolates. Restriction fragment length polymorphisms (RFLP) generated from whole-cell DNA digested with SfiI demonstrated the same RFLP pattern among outbreak isolates. Among comparison isolates, karyotyping distinguished some isolates that were indistinguishable by RFLP patterns. Karyotyping by PFGE appears to be the most useful molecular typing tool for discrimination among strains of C. rugosa and will be a useful marker for evaluating the epidemiology of future C. rugosa infections.

摘要

念珠菌属的医院感染已成为重症监护病房发病率和死亡率日益重要的原因。采用分子技术对一家医院先前记录的一组皱纹念珠菌感染中的10株皱纹念珠菌分离株(9株来自烧伤病房,1株来自另一个医院病房)以及从不同地理区域的一家转诊真菌检测实验室分离得到的8株皱纹念珠菌(对照分离株)进行了研究。分离株来自多个解剖部位。以18株皱纹念珠菌分离株为菌株同一性的标志物,对全细胞DNA进行脉冲场凝胶电泳(PFGE)。皱纹念珠菌分离株的PFGE核型显示有4至7条染色体带。核型分析显示,烧伤病房的9株暴发分离株具有相同的PFGE图谱,证实了克隆菌株的传播。来自另一个医院病房的分离株具有独特的核型。8株对照分离株显示出不同的PFGE核型模式。用SfiI消化全细胞DNA产生的限制性片段长度多态性(RFLP)显示,暴发分离株之间具有相同的RFLP模式。在对照分离株中,核型分析区分了一些RFLP模式无法区分的分离株。通过PFGE进行核型分析似乎是区分皱纹念珠菌菌株最有用的分子分型工具,并且将成为评估未来皱纹念珠菌感染流行病学的有用标志物。

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