DETECTION OF SULFATE-REDUCING BACTERIA FROM VARIOUS ECOTOPES BY REAL-TIME PCR

Abstract

The study of detection effectiveness of sulfate-reducing bacteria (SRB) in the samples from various ecotopes by microbiological (cultural by serial dilutions) and molecular biological (by real-time PCR) methods with designed test-systems lyophilized on the silicon microchip was performed. The developed DSRM and SRB2 test-systems for detection of the functional gene dsrA presence, encoding one of the key enzyme of dissimilatory sulphate-reduction pathway — dissimilatory sulfite reductase were used. It was found that the minimal determined SRB titres in water samples were 104 cells/ml and in soil samples they were 102–105 cells/g of absolutely dry soil. In natural and man-caused samples, the amount of SRB detected by the microbiological method was correlated with calculated values determined by the molecular biological method, Pearson’s indexes were r = 0.41–0.69 (k = 11, P ≤ 0.01–0.05). Thus, real-time PCR assay with designed test systems lyophilized on the silicon microchips is a high-quality and rapid method for the detection of SRB in various natural and man-caused ecotopes.

Keywords

• dsra gene
• test-systems
• sulfate-reducing bacteria
• biocorrosion
• real-time pcr.