Biologia, Bratislava 56/6: 729-738, 2002.

ISSN 0006-3088 (Biologia). ISSN 1335-6399 (Biologia. Section Cellular and Molecular Biology).

 

Full Paper

Comparison of MRSA-Screen latex agglutination, conventional phenotypic methods and mecA gene detection for identification of oxacillin resistance in staphylococci.

 

Vaclav Hajek1, Roman Pantucek2, Milan Kolar1*, Jiri Doskar2 & Stanislav Rosypal2

1 Institute of Medical Microbiology, Faculty of Medicine, Palacky University, 77515 Olomouc, Czech Republic; tel.: ++ 420 68 5632407, fax: ++ 420 68 5632966, e-mail: kolar@tunw.upol.cz

2 Department of Genetics and Molecular Biology, Faculty of Science, Masaryk University, 61137 Brno, Czech Republic

* corresponding author

Received: December 18, 2001 / Accepted: July 1, 2002

 

Abstract

On the whole, 128 staphylococcal isolates  (Staphylococcus aureus 65, Staphylococcus epidermidis 42, and Staphylococcus haemolyticus 21 strains) from patients of ten Czech hospitals were classified as susceptible or resistant to oxacillin by the standard broth dilution micro-method (using the MIC breakpoint of ≥4.0 mg/L for resistance). The MRSA-Screen test (Denka Seiken Co., Japan), a latex agglutination for the detection of penicillin binding protein PBP2a in oxacillin/methicillin-resistant staphylococci, was compared with other commonly used phenotypic susceptibility tests. The PCR detection of the mecA gene, the presence of which is equaled to oxacillin resistance, was used as a reference genotypic method. In addition, the determination of the femA gene was performed also by this procedure. All 68 isolates (S. aureus 26, S. epidermidis 21, and S. haemolyticus 21 strains), determined as oxacillin-resistant, contained the mecA gene. The femA gene was present in all these S. aureus strains. The other 60 isolates (S. aureus 39, and S. epidermidis 21 strains) were characterized as oxacillin-susceptible. All the S. aureus strains lacked the mecA gene, however, all but two were femA gene-positive. The examination of S. epidermidis strains demonstrated the mecA gene in six (29 %) of them. Most of these discrepancies between the mecA PCR tests and determination of the oxacillin MICs could be avoided by decreasing of the resistance breakpoint (≥ 0.5 mg/L) according to the new recommendation of the NCCLS from 1999. In this connection, however, other phenotypic susceptibility methods (MRSA-Screen, oxacillin-agar screen, and disk diffusion tests) should also be modified, as most of them give false negative results in the scope of the oxacillin MICs from 0.5 to 2.0 mg/L. When evaluating the MRSA-Screen test by 3 min, only 46 (68 %) oxacillin-resistant strains agglutinated, however, other 20 (29 %) strains (mostly coagulase-negative ones) were positive after prolongation of the reaction time from 3 to 9 min. The sensitivities and specificities for MRSA-Screen latex agglutination in comparison with the broth dilution micro-method, oxacillin agar screen assay, and disk diffusion test were as follows: 98.5 and 100 %, 98.5 and 100 %, 100 and 98 %, 100 and 97 %, respectively. These results showed that the MRSA-Screen test has a very good sensitivity and specificity for the detection of oxacillin resistance as compared with other conventional phenotypic procedures, however, its priorities are simplicity and speed as required for routine microbiological laboratories.

 

Key words: staphylococci, oxacillin resistance, MRSA-Screen test, mecA gene detection.