aureus isolates with lukS gene was a vital step for appropriate therapy and controlling the dissemination of this potentially virulent pathogen. In Malaysia, the presence of MRSA has been reported [37, 38], and cases of MRSA infection and colonization have also been reported in the neighboring countries of Singapore and Indonesia [32, 33]. The presented pentaplex PCR assay is robust and practicable Selleck OICR-9429 for culture
confirmation purposes. However, the 104 CFU/mL analytical sensitivity of this current pentaplex PCR assay might not sensitive enough for the direct testing of clinical specimens. A previous study by Gosbell et al, in 2001 confirmed that MRSA-screen test gave excellent sensitivity and specificity for MRSA detection, and was quicker and cheaper than PCR , while other study showed lower sensitivity and specificity in detecting methicillin resistance in CoNS  and couldn’t Selleckchem Target Selective Inhibitor Library identify neither PVL toxin encoding gene among staphylococci nor differentiate between CA-MRSA and HA-MRSA. Hence the PCR assay developed in the
present study will be useful in the epidemiological screening of MRSA patients or carriers. We are currently evaluating this assay for screening for MRSA carriage in Malaysia. Conclusion The PCR assay was able to detect four genes that are essential for the identification of S. aureus and its methicillin-resistant genotypes simultaneously in a single test within 4 h. The built-in internal control in this assay prevented false-negative results. The diagnostic accuracy was determined using 230 clinical specimens and showed 97.6% of sensitivity and 99.3% of specificity in detecting methicillin-resistant staphylococci. Hence, this test can be used as an effective diagnostic and surveillance tool to monitor the spread and emergence of MRSA. Methods Study design This was a cross-sectional study in which Fossariinae the retrospective sample size was calculated by using PS software (Dupont & Plummer, 1997) using Dichotomous based on the sensitivity of the E-test and PCR at 100% and 98%
respectively [41, 42]. The Research and Ethics Committee, School of Medical Sciences, Universiti Sains Malaysia, approved the study protocol. Bacterial strains and clinical specimens The Staphylococcus spp. reference strains and other bacteria used in this study are listed in Table 1. A total of 230 retrospective Staphylococcus spp. that were isolated from routine clinical specimens obtained from Hospital Universiti Sains Malaysia, from March 2006 to February 2007, were used in this study. Among the 230 clinical isolates, 86 were from nasal samples, 45 from blood samples, 34 from pus samples, 19 each from body fluid, wounds and CSF samples, and eight from urine samples. Screening of Staphylococcus spp. from clinical specimens by the 17-AAG solubility dmso conventional method The clinical isolates were inoculated onto Columbia blood agar (Merck, NJ, USA) plates with 5% sheep blood for 24 h at 37°C.