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

Palka-Santini, Maria; Pützfeld, Stefan; Cleven, Berit E.E.; Krönke, Martin; Krut, Oleg

doi:10.1016/j.mimet.2006.10.004

Cited by 28 publications

(8 citation statements)

References 56 publications

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“…Multiplex PCR technology involves adding various specific primers in the same PCR system to simultaneously detect multiple pathogens or resistance genes (2,10). With regard to microarray technology, a large number of probes must be fixed onto a support to detect and analyze a variety of sample sequences (11). Since numerous pathogens cause intracranial infection, the two technologies were combined in the present study to detect four types of bacteria and six resistance genes.…”

Section: Discussionmentioning

confidence: 99%

Application of microarray technology for the detection of intracranial bacterial infection

Shen

Guan

Zhang

et al. 2013

Experimental and Therapeutic Medicine

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The aim of this study was to assess the value of microarray technology for the detection of intracranial bacterial infection. A small gene chip was prepared based on the four pathogens commonly known to cause intracranial infection and the corresponding six types of common resistance genes in The Affiliated Hospital of Nantong University and The Affiliated Haian People’s Hospital of Nantong University. Cerebrospinal fluid samples were then collected from 30 patients with clinically diagnosed intracranial infection for the detection of the bacteria and resistance genes. The results were compared with the bacterial culture and sensitivity test results from the Department of Clinical Laboratories. The laboratory bacterial culture took 4–5 days, and revealed that 12 cases were positive and 18 cases were negative for bacteria. The microarray analysis took 1 day, and bacteria and resistance genes were detected in 15 cases. The 16S gene and drug resistance genes were detected in 8 cases; however, the bacterial strain was not identified. Seven cases appeared negative for bacteria and resistance genes. Microarray technology is rapid, sensitive and suitable for use in the detection of intracranial infections and other diseases for which conventional bacterial culture has a low positive rate.

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Section: Discussionmentioning

confidence: 99%

Application of microarray technology for the detection of intracranial bacterial infection

Shen

Guan

Zhang

et al. 2013

Experimental and Therapeutic Medicine

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“…Some prototype microarrays have already been described and applied to the diagnosis of sepsis (27,146,182). One of them also included six antibiotic resistance genes, but it was limited to the detection of Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli in positive blood cultures (27).…”

Section: Nat-based Assays For Detection and Identification Ofmentioning

confidence: 99%

The Era of Molecular and Other Non-Culture-Based Methods in Diagnosis of Sepsis

et al. 2010

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SUMMARY Sepsis, a leading cause of morbidity and mortality throughout the world, is a clinical syndrome with signs and symptoms relating to an infectious event and the consequent important inflammatory response. From a clinical point of view, sepsis is a continuous process ranging from systemic inflammatory response syndrome (SIRS) to multiple-organ-dysfunction syndrome (MODS). Blood cultures are the current “gold standard” for diagnosis, and they are based on the detection of viable microorganisms present in blood. However, on some occasions, blood cultures have intrinsic limitations in terms of sensitivity and rapidity, and it is not expected that these drawbacks will be overcome by significant improvements in the near future. For these principal reasons, other approaches are therefore needed in association with blood culture to improve the overall diagnostic yield for septic patients. These considerations have represented the rationale for the development of highly sensitive and fast laboratory methods. This review addresses non-culture-based techniques for the diagnosis of sepsis, including molecular and other non-culture-based methods. In particular, the potential clinical role for the sensitive and rapid detection of bacterial and fungal DNA in the development of new diagnostic algorithms is discussed.

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“…Nucleic acid amplification methods can provide same-day results once the blood cultures are positive for microbial growth; however, the majority of presently described assays are not designed to differentiate between MSSA and MRSA. Additionally, those nucleic acid amplification procedures that can differentiate between MSSA and MRSA often require labor-intensive protocols, technologically advanced equipment, and a colony isolated from subculture, or such assays are not readily available (14,17,28,30,33,37,42). The detection of mecA or MRSA in positive blood culture bottles using molecular methods has previously been very successful (4,36), but there are few reports on the rapid differentiation between MSSA and MRSA among other gram-positive organisms causing bacteremia (29,34,37).…”

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confidence: 99%

Clinical Validation of the Molecular BD GeneOhm StaphSR Assay for Direct Detection of Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus in Positive Blood Cultures

et al. 2007

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The rapid detection of Staphylococcus aureus bacteremia and a swift determination of methicillin susceptibility has serious clinical implications affecting patient mortality. This study evaluated the StaphSR assay (BD GeneOhm, San Diego, CA), a real-time PCR assay, for the identification and differentiation of methicillin-susceptible S. aureus (MSSA) and methicillin-resistant S. aureus (MRSA) from 300 positive blood cultures. The BD GeneOhm StaphSR assay was performed and interpreted according to the manufacturer's recommendations. Positive blood cultures (containing predominantly gram-positive cocci in clusters) were subcultured on 5% sheep blood agar plates. After 18 to 24 h of incubation, isolates morphologically consistent with S. aureus were presumptively identified by latex agglutination (Staphaurex Plus; Remel, Lenexa, KS). Susceptibility testing was initially performed with the Phoenix automated microbiology system (BD Diagnostics, Sparks, MD). Additional susceptibility testing of samples with discrepant results was done using BBL oxacillin screen agar (BD Diagnostics, Sparks, MD), oxacillin and cefoxitin Etests (AB Biodisk, Solna, Sweden) on Mueller-Hinton agar, an immunoassay for penicillin binding protein 2 (Denka Seiken Co., Tokyo, Japan), and mecA PCR. The sensitivity, specificity, and positive and negative predictive values of the BD GeneOhm StaphSR assay for MSSA detection were 98.9, 96.7, 93.6, and 99.5%, respectively. For the detection of MRSA, the BD GeneOhm StaphSR assay was 100% sensitive and 98.4% specific; positive and negative predictive values for MRSA detection were 92.6 and 100%, respectively. Inhibition was seen with only one sample, and the issue was resolved upon retesting. The BD GeneOhm StaphSR assay appears to be a valuable diagnostic tool for quickly differentiating bacteremia caused by MSSA and MRSA from that caused by other gram-positive cocci.

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

Cited by 28 publications

References 56 publications

Application of microarray technology for the detection of intracranial bacterial infection

Application of microarray technology for the detection of intracranial bacterial infection

The Era of Molecular and Other Non-Culture-Based Methods in Diagnosis of Sepsis

Clinical Validation of the Molecular BD GeneOhm StaphSR Assay for Direct Detection of Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus in Positive Blood Cultures

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