Authors : Mohammad Shafiq Zahari, Siti Nazrina Camalxaman, Mohd Nazri Abu, Wan Shahriman Yushdie, Wan Yusoff, Evana Kamarudin, Azlin Sham Rambely and Mohd Fahmi Mastuki
Abstract: Although, molecular detection has been found to be sensitive and specific but DNA extraction step is still essential, which is known to be time-consuming, costly and a high risk of contamination. Y-PER reagent had previously demonstrated its effectiveness in disruption of some bacterial cells for releasing genomic DNA with an extremely simple and rapid procedure. We therefore want to investigate if this reagent and technique could be applied on six common nosocomial pathogens including Escherichia coli, Staphylococccus aureus, Streptococcus pneumoniae, Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter spp. Two extraction methods were performed using boiling and Y-PER reagent technique and the resulting genomic DNA extractions were then subjected to PCR analysis by real-time multiplex PCR assay. DNA templates extracted by boiling method has shown a successful amplification for all of the six bacteria, while Y-PER technique only shows a positive amplification for Escherichia coli and Streptococcus pneumonia. Although, Y-PER method has a potential to be rapid and simple molecular diagnostic tool for releasing genomic DNA for PCR analysis, further optimization need to be done for many bacteria in order to fully utilize Y-PER as replacement for boiling method.
Mohammad Shafiq Zahari, Siti Nazrina Camalxaman, Mohd Nazri Abu, Wan Shahriman Yushdie, Wan Yusoff, Evana Kamarudin, Azlin Sham Rambely and Mohd Fahmi Mastuki, 2016. The Effectiveness of Yeast Protein Extraction Reagent (Y-Per) in Releasing Genomic DNA of Nosocomial Pathogens for Rapid Detection. Journal of Engineering and Applied Sciences, 11: 2683-2687.