Quantitative detection of the oil-degrading bacterium Acinetobacter sp. strain MUB1 by hybridization probe based real-time PCR
| Autor: | Prayad Pokethitiyook, Günter Korge, Suchart Upatham, Maleeya Kruatrachue, Sudarat Suanjit, Annemarie Hofmann, Seksan Phrommanich, Suksiri Vichasri Grams |
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| Rok vydání: | 2006 |
| Předmět: |
Detection limit
Strain (chemistry) Acinetobacter Hybridization probe Biology biology.organism_classification Microbiology Molecular biology DNA extraction Polymerase Chain Reaction law.invention Standard curve Real-time polymerase chain reaction Biodegradation Environmental Petroleum law Computer Systems Genes Bacterial Cytochrome P-450 CYP4A DNA Probes Polymerase chain reaction Soil Microbiology |
| Zdroj: | Microbiological research. 164(4) |
| ISSN: | 1618-0623 |
| Popis: | Quantitative detection of the oil-degrading bacterium Acinetobacter sp. strain MUB1 was performed using the SoilMaster() 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+/-3.0x10(6) and 5.4+/-3.0x10(2)CFUml(-1) cell suspension. The detection limit was about 540CFUml(-1), 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+/-3.0x10(4)CFUg(-1) 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. |
| Databáze: | OpenAIRE |
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