Rapid identification, virulence analysis and resistance profiling of Staphylococcus aureus by gene segment-based DNA microarrays:: Application to blood culture post-processing

被引:14
作者
Palka-Santini, Maria
Puetzfeld, Stefan
Cleven, Berit E. E.
Kroenke, Martin
Krut, Oleg
机构
[1] Univ Cologne, Ctr Med, Inst Med Microbiol Immunol & Hyg, D-50935 Cologne, Germany
[2] Univ Cologne, Ctr Med, Ctr Mol Med Cologne, D-50935 Cologne, Germany
关键词
bacteremia; molecular diagnostics; virulence factors; antibiotic resistance; strain genotyping; COAGULASE-NEGATIVE STAPHYLOCOCCI; IN-SITU HYBRIDIZATION; OLIGONUCLEOTIDE-MICROARRAY; METHICILLIN RESISTANCE; STREAM INFECTIONS; PCR; BACTEREMIA; ASSAY; EPIDEMIOLOGY; EXPRESSION;
D O I
10.1016/j.mimet.2006.10.004
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Up to now, blood culturing systems are the method of choice to diagnose bacteremia. However, definitive pathogen identification from positive blood cultures is a time-consuming procedure, requiring subculture and biochemical analysis. We developed a microarray for the identification of Staphylococcus aureus comprising PCR generated gene-segments, which can reduce the blood culture post-processing time to a single day. Moreover, it allows concomitant identification of virulence factors and antibiotic resistance determinants directly from positive blood cultures without previous amplification by PCR. The assay unambiguously identifies most of the important virulence genes such as tsst-1, sea, seb, eta and antibiotic resistance genes such as mecA, aacA-aphD, blaZ and ermA. To obtain positive signals, 20 ng of purified genomic S. aureus DNA or 2 mu g of total DNA extracted from blood culture was required. The microarray specifically distinguished S. aureus from Gram-negative bacteria as well as from closely related coagulase negative staphylococci (CoNS). The microarray-based identification of S. aureus can be accomplished on the same day blood cultures become positive in the Bactec (R). The results of our study demonstrate the feasibility of microarray-based systems for the direct identification and characterization of bacteria from cultured clinical specimens. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:468 / 477
页数:10
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