Abstract
In this work there was evaluated the method of detection of methicillin resistant Staphylococcus aureus (MRSA) by using two molecular and three phenotypic tests in investigation procedure of 70 strains of S.aureus isolated from animals. Recent findings of the new mecA homologue, mecALGA251, minimise the significance of mecA gene presence detection as a confirmation method of methicillin resistant Staphylococcus aureus identification. For this reason, along with multiplex PCR set of primers(165rDNK, nuc, mecA) for detection mecA gene, there was also used multiplex PCR set of primers (spa, mecA, pvl, mecALGA251) for differentiation mecALGA251 from mecA, with simultaneous detection of luk-PV and spa gene fragments. In all 70 investigated isolates there was detected the presence of specific 16 SrDNK fragment and nuc gene which encodes a thermostable S. aureus nuclease, while in 5 out of 70 S. aureus isolates, there was proven mecA gene presence using two multiplex PCR tests. In the investigated strains there was determined neither mecC (mecALGA251)gene presence, nor Panton Valentine Leukocidin encoding gene. By application cefoxitin disk-diffusion, latex-agglutination and two multiplex PCR tests, the identical results in identification 5 methicillin resistant out of 70 investigated S. aureus strains were obtained. In our investigation there was determined a complete correlation between the results of phenotypic and genotypic identification of methicillin resistant S. aureus.
Highlights
STAPHYLOCOCCUS AUREUS ISOLATED FROM ANIMALS
In this work there was evaluated the method of detection of methicillin resistant Staphylococcus aureus
three phenotypic tests in investigation procedure of 70 strains of S.aureus isolated from animals
Summary
Radosavljević V., Žutić Jadranka, Pavlović Ljiljana, Bošković Tamara, Radanović O., Žutić M.**. Vet. glasnik 68 (1-2) 89 - 99 (2014) Radosavljević V. i sar.: Metode detekcije i tipizacije meticilin-rezistentnih Staphylococcus aureus izolovanih iz životinja. Vet. glasnik 68 (1-2) 89 - 99 (2014) Radosavljević V. i sar.: Metode detekcije i tipizacije meticilin-rezistentnih Staphylococcus aureus izolovanih iz životinja vremenom detekcijom lukF-PV i spa genskih fragmenata, što omogućava i direktno spa tipiziranje sekvenciranjem PCR amplikona (Stegger i sar., 2012). Svih 70 sojeva S. aureus je ispitano na prisustvo produkta mecA gena, penicillin vezujućeg proteina (PBP2a) pomoću lateks aglutinacionog testa (Slidex MRSA Detection, bioMerieux). Za potvrdu rezistencije prema meticilinu detekcijom mecA gena i istovremeno utvrđivanje prisustva mecC (mecALGA251) gena, spa gena i Panton Valentin Leukocidin (PVL ili lukF-PV) kodirajućih gena, korišćen je multipleks PCR prema Stegger i sar. Veličina zona inhibicije oko diskova cefoksitina u disk difuzionoj metodi i rezultat PCR detekcije mecA gena
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