Open AccessReal-time PCR assay for detection of <em>Staphylococcus aureus</em>, Panton-Valentine Leucocidin and Methicillin Resistance directly from clinical samples
Author(s) -
Liliana Galia,
Marco Ligozzi,
Anna Bertoncelli,
Annarita Mazzariol
Publication year - 2019
Publication title -
aims microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.565
H-Index - 6
ISSN - 2471-1888
DOI - 10.3934/microbiol.2019.2.138
Subject(s) - staphylococcus aureus , microbiology and biotechnology , panton–valentine leukocidin , context (archaeology) , methicillin resistant staphylococcus aureus , coagulase , sccmec , real time polymerase chain reaction , biology , leukocidin , virulence , polymerase chain reaction , staphylococcus , virology , medicine , bacteria , gene , paleontology , biochemistry , genetics
Rapid detection of Methicillin Resistant Staphylococcus aureus (MRSA) is an important concern for both treatment and implementation of infection control policies. The present study provides an 'in house' real-time PCR assay to detect directly nuc , pvl , and mecA genes. The assay is able to perform identification of MRSA, Methicillin-Sensitive S. aureus , Methicillin-Resistant Coagulase Negative Staphylococci and the Panton-Valentine leukocidin virulence gene from rectal and pharyngeal swab samples in a screening context. We found an analytical sensitivity of this current Triplex PCR assay of 514 CFU/mL. Analytical specificity was tested with different Gram-positive and Gram-negative species and yielded no false-positive PCR signal. The sensitivity and specificity of the Triplex Real Time PCR were both 100% for these targets when compared with the culture and conventional methods. This assay is readily adaptable for routine use in a microbiology laboratory, as it will enable the implementation of timely and properly guided therapy and infection control strategies.