两种方法分离食品中葡萄球菌的鉴定

I. Regecová, J. Výrostková, František Zigo, M. Pipová, P. Jevinová, S. Demjanová
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摘要

从阿拉斯加鳕鱼肉(Theragra chalcogramma)、大西洋鲭鱼肉(Scomber scombrus)、大西洋鲱鱼肉(Clupea harengus)以及野鸡(Phasianus colchicus)、野兔(Oryctolagus cuniculus)和布林德扎奶酪的股肌样品中分离出193株葡萄球菌。在这些分离株中检测到单个葡萄球菌物种的典型表型表现。采用基质辅助激光解吸电离质谱法对葡萄球菌进行物种鉴定,鉴定出金黄色葡萄球菌、表皮葡萄球菌、头葡萄球菌、溶血葡萄球菌、沃纳葡萄球菌5种。随后,通过PCR检测证实了16S rDNA基因的存在;金黄色葡萄球菌的特定序列;利用sodA基因的特异序列ser0107作为推测的转录调控基因,对表皮葡萄球菌、瓦尔纳氏葡萄球菌、溶血葡萄球菌和头孢葡萄球菌进行了鉴定。将PCR法和基质辅助激光解吸电离质谱法鉴定结果进行比较,所有病例在属水平上的鉴定结果相同(100%)。然而,结果的差异在物种水平上得到证实,其中PCR检测和基质辅助激光解吸电离质谱鉴定的一致性仅为80%。分离株的得分范围为1.953 ~ 2.564。
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IDENTIFICATION OF STAPHYLOCOCCUS SPP. ISOLATED FROM FOOD BY TWO METHODS
One hundred and ninety-three staphylococcal strains were isolated from samples of alaska pollock meat (Theragra chalcogramma), atlantic mackerel meat (Scomber scombrus),  atlantic herring meat (Clupea harengus), and femoral muscle samples of the wild pheasants (Phasianus colchicus), wild rabbits (Oryctolagus cuniculus) and bryndza cheese. Phenotypic manifestations typical of individual staphylococcal species were detected in these isolates. Species identification was also performed by matrix-assisted-laser-desorption-ionization-mass-spectrometry on the basis of which 5 species of staphylococci were determined: S. aureus, S. epidermidis, S. capitis, S. haemolyticus, S. warneri. Subsequently, the presence of the 16S rDNA gene was confirmed by PCR assay; a specific sequence for S. aureus species; putative transcriptional regulator gene serp0107, a specific sequence of the sodA gene was used to identify S. epidermidis and to identify S. warneri, S. haemolyticus, S. capitis species. When comparing the results of identification by PCR assay and matrix-assisted-laser-desorption-ionization-mass-spectrometry, the identification at the genus level was identical in all cases (100%). However, the discrepancies in results were confirmed at the species level, where congruence for identification of isolates by PCR assay and matrix-assisted-laser-desorption-ionization-mass-spectrometry was only 80%. For these isolates, the score value ranged from 1.953 to 2.564.
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