A Novel Approach Based on Real-Time PCR with High-Resolution Melting Analysis for the Simultaneous Identification of Staphylococcus aureus and Staphylococcus argenteus .

Autor: Chieffi D; Institute of Sciences of Food Production, National Research Council of Italy (CNR-ISPA), 70126 Bari, Italy., Bongiorno D; Medical Molecular Microbiology and Antibiotic Resistance Laboratory (MMARLab), Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, 95123 Catania, Italy., Licitra A; Institute of Sciences of Food Production, National Research Council of Italy (CNR-ISPA), 70126 Bari, Italy., Campanile F; Medical Molecular Microbiology and Antibiotic Resistance Laboratory (MMARLab), Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, 95123 Catania, Italy., Fusco V; Institute of Sciences of Food Production, National Research Council of Italy (CNR-ISPA), 70126 Bari, Italy.
Jazyk: angličtina
Zdroj: Foods (Basel, Switzerland) [Foods] 2024 Sep 22; Vol. 13 (18). Date of Electronic Publication: 2024 Sep 22.
DOI: 10.3390/foods13183004
Abstrakt: Staphylococcus ( S .) aureus is a pathogenic bacterium able to cause several diseases in humans and animals as well as foodborne intoxications. S. argenteus , being phenotypically and genotypically related to S. aureus , is part of the so-called S. aureus complex and recently recognized as an emerging pathogen able to cause, like S. aureus , several diseases both in humans and animals, and foodborne poisoning outbreaks. However, it has been reported that the widely used conventional PCR of Brakstad et al. [ Journal of Clinical Microbiology , 30 (7), 1654-1660, (1992)] targeting the thermostable nuclease gene may provide false-positive S. aureus , as it is able to amplify also S. argenteus . Here, we developed a novel two-step approach that, following the PCR of Brakstad et al. (1992), discriminates S. aureus from S. argenteus by a real-time PCR with high-resolution melting analysis (rt-PCR-HRM). In particular, targeting a polymorphic 137 bp region of the sodA gene, our developed rt-PCR-HRM method clearly discriminated S. aureus from S. argenteus , showing a remarkable difference in their amplification product melting temperatures (approximately 1.3 °C) as well as distinct melting curve shapes. The good sensitivity, reproducibility, user friendliness, and cost effectiveness of the developed method are advantageous attributes that will allow not only its easy employment to correctly identify misidentified isolates present in various collections of S. aureus, but also expand the still lacking knowledge on the prevalence and distribution of S. argenteus .
Databáze: MEDLINE