Multiplex PCR Assay for Simultaneous Detection of Nine Clinically Relevant Antibiotic Resistance Genes inStaphylococcus aureus

Abstract

In this study we describe a multiplex PCR assay for the detection of nine clinically relevant antibiotic resistance genes of Staphylococcus aureus. Conditions were optimized to amplify fragments of mecA (encoding methicillin resistance), aacA-aphD (aminoglycoside resistance), tetK, tetM (tetracycline resistance), erm(A), erm(C) (macrolide-lincosamide-streptogramin B resistance), vat(A), vat(B), and vat(C) (streptogramin A resistance) simultaneously in one PCR amplification. An additional primer pair for the amplification of a fragment of the staphylococcal 16S rDNA was included as a positive control. The multiplex PCR assay was evaluated on 30 different S. aureus isolates, and the PCR results correlated with the phenotypic antibiotic resistance data obtained by the broth microdilution assay. The multiplex PCR assay offers a rapid, simple, and accurate identification of antibiotic resistance profiles and could be used in clinical diagnosis as well as for the surveillance of the spread of antibiotic resistance determinants in epidemiological studies.

Correlation Between the Genotype Detection Determined by Rt- PCR Assays and the Phenotypic Detection Determined by Antibiotic Susceptibility Patterns of Methicillin-Resistant Staphylococcus Aureus Clinicalisolates in Algeria.

“…Overall, we found high correlation between these two methods. This correlation has been demonstrated previously in other studies [6,14,15,17].…”

“…In recent years, several studies have demonstrated the high capacity of molecular methods such as Pulsed-field gel electrophoresis (PFGE), RT-PCR and PCR-based methods to increase the rapidity and accuracy of resistance testing [1,6,14,15,16,21,22].…”

“…5(5), 950-955 951 Since then, the resistance rate of methicillin-resistant S. aureus (MRSA) has gradually increased and become a serious problem in both community and hospital infections worldwide in 1970 [12]. Until the mid-1990s, the majority of MRSA isolates exhibited multiple resistance to several antimicrobial agents including tetracyclines, aminoglycosides, macrolides, lincosamides and fluoroquinolones [5,12,15,16].…”

Antimicrobial Resistance of Coagulase-negative Species of Staphylococci Isolated from the Meat of Wild Pheasants (Phasianus Colchicus)

“…The total genomic DNA was isolated from staphylococcal strains as described by Hein et al (2005) and further tested by polymerase chain reaction (PCR) method according to Strommenger et al (2003). Primers 16S1 (5-CAGCTCGTGTCGTGAGATGT) and 16S2 (5-AATCATTTGTCCCACCTTCG) rDNA (Generi Biotech s.r.o., Hradec Králové, Czech Republic) typical for the genus Staphylococcus were used for PCR.…”

A Fatal Case of Infective Endocarditis Caused by Community-Associated Methicillin-ResistantStaphylococcus aureusST72 in Korea

“…To determine whether the isolate would have the characteristic features of CA-MRSA, we conducted genetic studies. The detection of mecA gene and typing of staphylococcal cassette chromosome mec (SCCmec) were performed separately using PCR methods with each specific primer for mecA [4] and for SCCmec [5]. The isolate had mecA gene and SCCmec type IV A (Fig.…”