Nikolina RUSENOVA, Svetozar KRUSTEV, Anatoli ATANASOV, Anton RUSENOV, Spaska STANILOVA

Differentiation of Staphylococcus pseudintermedius in the Staphylococcus intermedius Group (SIG) by Conventional and Molecular Methods

Staphylococcus pseudintermedius is a member of the Staphylococcus intermedius group (SIG) comprising representatives with similar phenotypic characteristics. The purpose of this study was to optimise a method for accurate differentiation of S. pseudintermedius in clinical samples from dogs and cats based on PCR-RFLP analysis, following the key biochemical tests for SIG group assignment, and to determine the prevalence of S. pseudintermedius compared to other members of the SIG group. To implement this, 158 isolates from dogs and 17 from cats, 4 reference strains - S. intermedius ATCC 29663, S. pseudintermedius LMG 22219, Staphylococcus delphini ATCC 49171, Staphylococcus aureus ATCC 25922 and 5 field strains of S. aureus were analysed. The generated PCR-RFLP profile (pta gene, MboI restriction enzyme) clearly differentiated S. pseudintermedius within the SIG group by having two distinctive bands of 213 and 107 bp, respectively. The data showed a predominance of S. pseudintermedius in clinical samples from dogs, while no S. pseudintermedius were found among the cat’s samples. In conclusion, due to the atypical biochemical reactions found in some S. pseudintermedius isolates (14.6%), precise identification should be based on an extensive phenotypic assay together with a molecular biological method or the latter alone.

Staphylococcus intermedius Grubundaki (SIG) Staphylococcus pseudintermedius’un Geleneksel ve Moleküler Yöntemlerle Ayırdedilmesi

Staphylococcus pseudintermedius, benzer fenotipik özelliklere sahip temsilciler içeren Staphylococcus intermedius grubunun (SIG) bir üyesidir. Bu çalışmanın amacı, köpek ve kedilerden elde edilen klinik örneklerde SIG grubu ataması için temel biyokimyasal testleri takiben PCR-RFLP analizine dayanarak S. pseudintermedius’un doğru şekilde ayırdedilmesi için bir yöntemi optimize etmek ve SIG grubunun diğer üyelerine kıyasla S. pseudintermedius’un yaygınlığını belirlemekti. Bu amaçla, köpeklerden elde edilen 158 ve kedilerden elde edilen 17 izolat, 4 referans suş - S. intermedius ATCC 29663, S. pseudintermedius LMG 22219, Staphylococcus delphini ATCC 49171, Staphylococcus aureus ATCC 25922 ve S. aureus’un 5 saha suşu analiz edildi. Oluşturulan PCR-RFLP profili (pta geni, MboI kısıtlama enzimi), S. pseudintermedius’u sahip olduğu 213 ve 107 bp’lik iki farklı bantla SIG grubu içinde belirgin şekilde ayırdetti. Veriler, köpeklerden alınan klinik örneklerde S. pseudintermedius’un baskın olduğunu gösterirken, kedi örneklerinde S. pseudintermedius belirlenmedi. Sonuç olarak, bazı S. pseudintermedius izolatlarında bulunan atipik biyokimyasal reaksiyonlar (%14.6) nedeniyle kesin tanı, kapsamlı bir fenotipik test ile birlikte kullanılacak moleküler biyolojik bir yöntem veya sadece moleküler yönteme dayanmalıdır.

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