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Title: Quantitative detection of the oil-degrading bacterium Acinetobacter sp. strain MUB1 by hybridization probe based real-time PCR
Authors: Seksan Phrommanich
Sudarat Suanjit
Suchart Upatham
Suksiri Vichasri Grams
Maleeya Kruatrachue
Prayad Pokethitiyook
Günter Korge
Annemarie Hofmann
Burapha University
Mahidol University
Freie Universitat Berlin
Keywords: Immunology and Microbiology
Issue Date: 1-Jan-2009
Citation: Microbiological Research. Vol.164, No.4 (2009), 486-492
Abstract: Quantitative detection of the oil-degrading bacterium Acinetobacter sp. strain MUB1 was performed using the SoilMaster<sup>™</sup> DNA Extraction Kit (Epicentre, Madison, Wisconsin) and hybridization probe based real-time PCR. The detection target was the alkane hydroxylase gene (alkM). Standard curve construction showed a linear relation between log values of cell concentrations and real-time PCR threshold cycles over five orders of magnitude between 5.4±3.0×10<sup>6</sup> and 5.4±3.0×10<sup>2</sup> CFU ml<sup>-1</sup> cell suspension. The detection limit was about 540 CFU ml<sup>-1</sup>, which was ten times more sensitive than conventional PCR. The quantification of Acinetobacter sp. strain MUB1 cells in soil samples resulted in 46.67%, 82.41%, and 87.59% DNA recovery with a detection limit of 5.4±3.0×10<sup>4</sup> CFU g<sup>-1</sup> dry soil. In this study, a method was developed for the specific, sensitive, and rapid quantification of the Acinetobacter sp. strain MUB1 in soil samples. © 2007 Elsevier GmbH. All rights reserved.
ISSN: 09445013
Appears in Collections:Scopus 2006-2010

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