Rapid Diagnosis of Methicillin-Resistant Staphylococcus aureus Bacteremia by Nested Polymerase Chain Reaction

Abstract

ObjectiveThe purpose of this study was to establish a rapid and sensitive diagnostic method for methicillinresistant Staphylococcus aureus (MRSA) bacteremia in postoperative patients. Summary Background DataAs a result of diffusion and abuse of third-generation cephalosporin antibiotics in the 1980s in Japan, an outbreak of MRSA infection has been posed. In the field of surgery, severe postoperative infections with MRSA such as MRSA bacteremia, which may lead to multiple organ failure, have emerged with a high mortality. MethodsThirty-five patients with high fever (above 38.5 C) or watery diarrhea or both within 2 weeks after gastrointestinal major surgery and 6 healthy volunteers were examined. Nested polymerase chain reaction was used to detect mecA and toxic shock syndrome toxin-1 (TSST-1) genes in blood specimens. ResultsThe mecA and TSST-1 genes were not detected in the blood samples of any of the six healthy volunteers. In all 12 samples from which MRSA colonies were isolated by blood culture, mecA and TSST-1 genes were detected. Although it took at least 48 hours to identify MRSA by the blood culture method, the presence of mecA and TSST-1 genes was determined by nested polymerase chain reaction method within only 3 to 4 hours after blood sampling. ConclusionsThis method, as a sensitive and rapid monitoring system for MRSA bacteremia, would be clinically beneficial for prevention of cross infection and for early determination of appropriate treatment for infected patients. 665

Molecular Detection of Antimicrobial Resistance

“…A limited study by Kitagawa et al (151) in Japan demonstrated the ability of PCR to directly detect MRSA in clinical specimen. This group developed a nested multiplex PCR for mecA and TSST-1 for use with 1-ml blood samples.…”

Polymerase Chain Reaction: Types, Utilities and Limitations

“…Contamination can be controlled using primers designed to anneal at different temperatures. Contamination can also be controlled by adding ultra-pure oil to make a physical separation of two mixtures of amplification [15,17,18]. …”

Bioinformatics Analysis Of Novel Non-Coding Motifs In Pathogenic Bacterial Genome

“…The main problems with these systems are the presence of inhibitors and the amplification of non-targets, which can give false negative results (Kasai et al 2000 andStrommenger et al 2003). Detection of MecA gene is also used in many hospitals which can be done by either by radio labeling, PCR amplification or simply by enrichment culture broths (Kitagawa et al 1996).But this method is not very well established as it can only detect nasal and wound infections (Brown et al 2005 andOberdorfer et al2006). Another DNA screening can be done to find the Panton-Valentine leukocidin (PVL) gene, which is also a toxin producer in MRSA infection (Shrestha et al 2002).…”