A highly selective direct method of detecting sulphate-reducing bacteria in crude oil

Abstract

Aims: The aim of this work was to develop a highly selective method of detecting sulphate-reducing bacteria (SRB) in crude oil. Methods: A pair of PCR primers was designed based on an alignment of the nucleotide sequence of the 16S rRNA genes from the Desulfovibrionaceae family. DNA extraction from crude oil was performed by the method using zirconia beads and a stool kit. Results: The PCR specifically detected Desulfovibrio and Desulfomicrobium in a sediment sample. When nucleic acids extracted directly from crude oil were used for the PCR, 16S rRNA genes of Desulfovibrio and Thermodesulforhabdus norvegicus were detected. Impact of study: A simple direct method for detection of the SRB in crude oil using PCR was established.