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Challenges with the Applicaion of qPCR Based Tests for Enterococci as a Rapid Beach Mangement Tool in Hawai'i

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Title:Challenges with the Applicaion of qPCR Based Tests for Enterococci as a Rapid Beach Mangement Tool in Hawai'i
Authors:Seruge, Jaline
Contributors:Kirs, Marek (advisor)
Oceanography (department)
Global Environmental Science (department)
Keywords:molecular biology
Date Issued:2017
Publisher:University of Hawaiʻi at Mānoa
Place of Publication:Honolulu
Abstract:Rapid quantitative PCR based methods (USEPA Method 1609 and 1611) that can
evaluate water quality within a few hours are important since urbanization and our
changing climate will alter inputs of bacteria into the environment, which can
compromise the health of Hawai’i residents and visitors. We identified that assay
interference in Hawaiian coastal waters is associated with the DNA extraction process,
but not with the PCR amplification procedure. Further we demonstrated that acidification
of water samples can alleviate assay interference associated with Hawai’i’s beach water
It is speculated that acidification of coastal water samples helped to dissolve
calcium carbonate rich coralline particles, common to subtropical and tropical coastal
waters, hence eliminating a substratum for DNA to bind. This study envisions, that once
the assay can be used to reliably quantify microorganisms (such as enterococci, humanassociated Bacteroides), these tests will be used for beach water quality evaluations at
popular beaches in the state of Hawai’i.
In this research, water matrixes for PCR inhibitors were tested from water
samples gathered at selected beaches on Oahu, by using the EPA Methods 1609 and
1611, Enterococci in Water by TaqMan ®Quantitative Polymerase Chain Reaction
(qPCR) with Internal Amplification Control (IAC) Assay. There is a relationship between
the sodium chloride and calcium carbonate on Oahu, and the inhibition of the rapid
method, used to identify enterococci. The corals in Hawai’i are unique and add in with
the water's composition that is interfering with the DNA extraction process, and
underestimating the rapid method results.
Pages/Duration:41 pages
Rights:All UHM dissertations and theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission from the copyright owner.
Rights Holder:Seruge, Jaline
Appears in Collections: Global Environmental Science Theses

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