Karina Yew-Hoong Gin
Department of Civil and Environmental Engineering, National University of Singapore,
Singapore
Research Article
The Effects of Pretreatments on qPCR Detection of Bacteria - Bacteria
Indicator, Enterococcus as a Model
Author(s): Shin Giek Goh and Karina Yew-Hoong GinShin Giek Goh and Karina Yew-Hoong Gin
Current recreational water guidelines (e.g. USEPA and WHO) recommend enterococci as the indicator for both freshwaters and marine waters. Culture-based method has been used for decades to quantify the presence of enterococci. In year 2012, USEPA guideline for recreational water included qPCR as the alternative method to detect enterococci. qPCR method does not require long incubation time as of culture-based method. In addition, it offers the advantages of high specificity and sensitivity detection. Nevertheless, qPCR hampered by three main limiting factors. There are limited target cell in small volume of sample being analyzed by qPCR, the presence of inhibitors that can easily affect the sensitive detection of qPCR and the persistence of free DNA (DNA released from dead cell) that caused the false-positive signal in qPCR. As such, the upstream treatments are crucial to the accuracy .. View More»
DOI:
10.4172/1948-5948.1000193