Real-Time PCR Method for the Rapid Detection and Quantification of Pathogenic <i>Staphylococcus</i> Species Based on Novel Molecular Target Genes
Coagulase-positive <i>Staphylococcus aureus</i> is a foodborne pathogen considered one of the causes of food-related disease outbreaks. Like <i>S. aureus</i>, <i>Staphylococcus capitis</i>, <i>Staphylococcus caprae</i>, and <i>S. epidermidis</...
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| Main Authors: | , , , , , |
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| Format: | article |
| Language: | EN |
| Published: |
MDPI AG
2021
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| Subjects: | |
| Online Access: | https://doaj.org/article/fb6b32542d5842ebb4a7fead2b6e46cb |
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| Summary: | Coagulase-positive <i>Staphylococcus aureus</i> is a foodborne pathogen considered one of the causes of food-related disease outbreaks. Like <i>S. aureus</i>, <i>Staphylococcus capitis</i>, <i>Staphylococcus caprae</i>, and <i>S. epidermidis</i> are opportunistic pathogens causing clinical infections and food contamination. The objective of our study was to develop a rapid, accurate, and monitoring technique to detect four <i>Staphylococcus</i> species in food. Four novel molecular targets (GntR family transcriptional regulator for <i>S. aureus</i>, phosphomannomutase for <i>S. epidermidis</i>, FAD-dependent urate hydroxylase for <i>S. capitis</i>, and Gram-positive signal peptide protein for <i>S. caprae</i>) were mined based on pan-genome analysis. Primers targeting molecular target genes showed 100% specificity for 100 non-target reference strains. The detection limit in pure cultures and artificially contaminated food samples was 10<sup>2</sup> colony-forming unit/mL for <i>S. aureus</i>, <i>S. capitis</i>, <i>S. caprae</i>, and <i>S. epidermidis</i>. Moreover, real-time polymerase chain reaction successfully detected strains isolated from various food matrices. Thus, our method allows an accurate and rapid monitoring of <i>Staphylococcus</i> species and may help control staphylococcal contamination of food. |
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