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Journal of Clinical Microbiology, July 2004, p. 3147-3152, Vol. 42, No. 7
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.7.3147-3152.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Evaluation of the Hyplex BloodScreen Multiplex PCR-Enzyme-Linked Immunosorbent Assay System for Direct Identification of Gram-Positive Cocci and Gram-Negative Bacilli from Positive Blood Cultures

Nele Wellinghausen,1* Beate Wirths,1 Andreas Essig,1 and Lars Wassill2

Department of Medical Microbiology and Hygiene, University of Ulm, Ulm,1 Amplex Diagnostics GmbH, Munich, Germany2

Received 28 January 2004/ Returned for modification 25 March 2004/ Accepted 31 March 2004

We evaluated the Hyplex BloodScreen PCR-enzyme-linked immunosorbent assay (ELISA) system (BAG, Lich, Germany), a new diagnostic test for the direct identification of gram-negative bacilli and gram-positive cocci from positive blood cultures, with 482 positive BACTEC 9240 blood culture bottles. The test involves amplification of the bacterial DNA by multiplex PCR and subsequent hybridization of the PCR product to specific oligonucleotide probes in an ELISA-based format. The available probes allow the separate detection of Escherichia coli, Pseudomonas aeruginosa, Enterobacter aerogenes, Klebsiella spp., Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis/Enterococcus faecium, Streptococcus pyogenes, and Streptococcus pneumoniae and the staphylococcal mecA gene. The Hyplex BloodScreen test showed an overall sensitivity of 100% for the identification of gram-negative bacilli and 96.6 to 100% for the identification of gram-positive cocci (S. aureus, 100%; S. epidermidis, 97.2%; Enterococcus faecalis/Enterococcus faecium, 96.6%; and Streptococcus pneumoniae, 100%). The specificities of the test modules ranged from 92.5 to 100% for gram-negative bacilli and 97.7 to 100% for gram-positive cocci (Escherichia coli, 92.5%; Pseudomonas aeruginosa, 98.5%; Klebsiella spp., 100%; Enterobacter aerogenes, 100%; S. aureus, 100%, S. epidermidis, 97.7%; Enterococcus faecalis/Enterococcus faecium, 99.6%; Streptococcus pyogenes, 100%; and Streptococcus pneumoniae, 99.3%). The result of the mecA gene detection module correlated with the result of the phenotypic oxacillin resistance testing in all 38 isolates of Staphylococcus aureus investigated. In conclusion, the Hyplex BloodScreen PCR-ELISA system is well suited for the direct and specific identification of the most common pathogenic bacteria and the direct detection of the mecA gene of Staphylococcus aureus in positive blood cultures.

* Corresponding author. Mailing address: Department of Medical Microbiology and Hygiene, University of Ulm, Robert-Koch-Str. 8, D-89081 Ulm, Germany. Phone: 49-(0)731-500-24603. Fax: 49-(0)731-500-23473. E-mail: nele.wellinghausen{at}medizin.uni-ulm.de.

Journal of Clinical Microbiology, July 2004, p. 3147-3152, Vol. 42, No. 7
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.7.3147-3152.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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