Design and application of a Staphylococcus-specific single strand conformation polymorphism-PCR analysis to monitor Staphylococcus populations diversity and dynamics during production of raw milk cheese

Abstract

Aim: Development of a nested-PCR single strand conformation polymorphism (SSCP) assay targeting the 16S rRNA genes of the Staphylococcus genus, to monitor staphylococci in cheese. Methods and Results: New primer sets to specifically amplify 16S rDNA of staphylococci were designed to be used in a nested-PCR SSCP assay. The method was efficient in discriminating the staphylococcal species most frequently found in cheese. It was validated by monitoring Staphylococcus populations in three productions of raw milk cheese. Analysis of milk samples revealed dominant SSCP peaks corresponding to Staphylococcus aureus, Staphylococcus equorum and Staphylococcus saprophyticus. After 12 h, the S. aureus peak became dominant. Conclusions: The combination of specific Staphylococcus nested-PCR and SSCP allows rapid and direct monitoring of staphylococci diversity and dynamics in milk and cheese. In the core of the cheeses studied, S. aureus may have ecological advantages against other Staphylococcus populations. Significance and Impact of the Study: This approach is a promising tool to study the ecology of staphylococci in cheeses and in other food samples.

Application of Different Molecular Techniques for Characterization of Catalase‐Positive Cocci Isolated from Sucuk

“…S. saprophyticus is a widespread species generally detected in several fermented meat products and various types of cheese, and even in the food processing environment due to its good adaptation capability (Delbès and Montel, ; Rea and others, ; Soares and others, ). S. saprophyticus has previously been reported to be the dominant species in dry‐fermented Greek sausages (Papamanoli and others ; Drosinos and others ), Argentinian sausages (Fontana and others ), and Iberian sausages (Benito and others ) or the codominant species in Italian‐type fermented sausages (Mauriello and others ).…”

Sources of the adventitious microflora of a smear-ripened cheese

“…1996; Irlinger et al. 1997; Delbès and Montel 2005) and from the surface of fermented meat products, where the curing process and the salt are considered to be the main sources of contamination (Vilar et al. 2000; Gardini et al.…”

Application of PCR-Based Methods to Dairy Products and to Non-Dairy Probiotic Products

“…Single-strand conformation polymorphism-PCR (SSCP-PCR) is another PCR-based method for microbial diversity investigation that has been applied to dairy products (Duthoit et al, 2003;Feurer et al, 2004a;Feurer et al, 2004b;Delbes and Montel, 2005;Duthoit et al, 2005;Callon et al, 2006;Delbes et al, 2007;Saubusse et al, 2007;Mounier et al, 2009). This technique is based on the sequence-dependent differential intra-molecular folding of single strand DNA, which alters the migration speed of the molecules under non-denaturing conditions.…”

“…The hypervariable regions are flanked by conserved sequences, which can serve for amplification with "universal" primers (Baker et al, 2003). The variable V1 Bonetta et al, 2008), V3 Ercolini et al, 2001;Ogier et al, 2002;Duthoit et al, 2003;Ercolini et al, 2003;Mauriello et al, 2003;Andrighetto et al, 2004;Ercolini et al, 2004;Feurer et al, 2004a;Feurer et al, 2004b;Lafarge et al, 2004;Ogier et al, 2004;Duthoit et al, 2005;Flórez and Mayo, 2006;Delbes et al, 2007;El-Baradei et al, 2007;Parayre et al, 2007;Abriouel et al, 2008;Ercolini et al, 2008;Gala et al, 2008;Van Hoorde et al, 2008;Alegría et al, 2009;Casalta et al, 2009;Dolci et al, 2009;Giannino et al, 2009;Mounier et al, 2009;Serhan et al, 2009;Dolci et al, 2010;Fontana et al, 2010;Van Hoorde et al, 2010;Masoud et al, 2011), V2 (Duthoit et al, 2003;Delbes and Montel, 2005;Saubusse et al, 2007), V4-V5 , V1-V3 (Randazzo et al, 2002), V4-V8 (Randazzo et al, 2006), V5-V6 (Le Bourhis et al, 2005;Le Bourhis et al, 2007) and V6-V8 …”