TY - JOUR
T1 - The Effectiveness of Yeast Protein Extraction Reagent (Y-Per) in Releasing Genomic DNA of Nosocomial Pathogens for Rapid Detection
AU - Fahmi Mastuki, Mohd AU - Shafiq Zahari, Mohammad AU - Nazrina Camalxaman, Siti AU - Nazri Abu, Mohd AU - Shahriman Yushdie, Wan AU - Yusoff, Wan AU - Kamarudin, Evana AU - Sham Rambely, Azlin
JO - Journal of Engineering and Applied Sciences
VL - 11
IS - 12
SP - 2683
EP - 2687
PY - 2016
DA - 2001/08/19
SN - 1816-949x
DO - jeasci.2016.2683.2687
UR - https://makhillpublications.co/view-article.php?doi=jeasci.2016.2683.2687
KW - Yeast Protein Extraction Reagent (Y-PER)
KW -Nosocomial pathogens
KW -SYBR green real-time PCR
KW -diagnostic
KW -molecular
AB - 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.
ER -