Detection of Nocardia sp. H17-1 by PCR during Bioremediation of Crude Oil-Contaminated Soil

Abstract

For the detection of the oil-degrading bacterium, Nocardia sp. H17-1, inoculated during the bioremediation of oil-contaminated soil, a species-specific primer was constructed based on the 16S rDNA sequence of this strain. Two forward primers and two reverse primers were designed and tested against both closely and distantly related bacterial strains. All the primers designed were specific to the Nocardia sp. H17-1. Particularly, primer sets NH169F-NH972R and NH575F-NH972R could be used to detect 50 fg of template DNA and 1.2×104 CFU/g of sandy soil. These two PCR primer sets successfully detected the H17-1 strain in the oil-contaminated soil samples containing heterogeneous DNA. We also conformed the primer specificity by restriction-enzyme cleavage of the PCR products and denaturing gradient gel electrophoresis.