Quantitative detection of the oil-degrading bacterium Acinetobacter sp. strain MUB1 by hybridization probe based real-time PCR.
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.
Phrommanich
Seksan
S
Biological Science Graduate Program, Faculty of Science, Burapha University, Bangsaen, Chonburi, Thailand. seksan55@yahoo.com
Suanjit
Sudarat
S
Upatham
Suchart
S
Grams
Suksiri Vichasri
SV
Kruatrachue
Maleeya
M
Pokethitiyook
Prayad
P
Korge
Günter
G
Hofmann
Annemarie
A
D000150:Acinetobacter / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D001673:Biodegradation, Environmental; D003199:Computer Systems; D042926:Cytochrome P-450 CYP4A / Q000235:genetics* / Q000378:metabolism; D015342:DNA Probes; D005798:Genes, Bacterial; D010578:Petroleum / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012988:Soil Microbiology
D015342:DNA Probes; D010578:Petroleum; D042926:Cytochrome P-450 CYP4A