10253 Detection of SRP Activity by Quantification of MRNA for the Dissimulatory (Bi) Sulfite Reductase Gene (DSRA) by Reverse Transciptasea Quantitative PCR

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Product Number: 51300-10253-SG

ISBN: 10253 2010 CP

Author: Andrew C. Price, Laura Acuna Alvarez, Corinne Whitby and Jan Larsen

Publication Date: 2010

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Molecular biological methods have been used for some years to identify and quantify active microorganisms present in a commercial oil reservoir where biogenic sulfide production is routinely controlled by nitrate injection. In order to gain a more complete understanding of the effects of nitrate injection on the activity of sulfate reducing prokaryotes (SRP, (which encompasses sulfate reducing Bacteria (SRB)) and sulfate reducing Archaea (SRA)), the mRNA for dsrA present in produced water samples was quantified by reverse transcription quantitative PCR (RT-qPCR); mRNA for dsrA should only be produced by SRP actively reducing sulfate. The aims of this study were: to help further our understanding on the mode of action of nitrate on SRP activity e.g. competitive inhibition by nitrate utilising Bacteria (NUB), nitrite toxicity, change in reduction-oxidation potential or a metabolism switch from sulfate to nitrate reduction, and; to provide a rapid monitoring tool for SRP activity.

Since messenger RNA is known to be unstable and is rapidly processed within cells, the first task was to design a laboratory experiment to demonstrate that mRNA for dsrA could be detected and quantified in produced water samples. Produced water samples were spiked with a SRP culture grown from the produced water sample and the mRNA for dsrA was successfully detected and quantified.

Keywords. dissimilatory (bi) sulfite reductase, dsrAB, dsrA, Halfdan oilfield, hydrogen sulfide (H2S), MIC, molecular microbiology methods (MMM), most probable number (MPN), nitrate injection, nitrate utilizing bacteria (NUB), quantitative polymerase chain reaction (qPCR), reservoir souring, reverse transcription quantitative polymerase chain reaction (RT-qPCR), sulfate-reducing Archaea (SRA), sulfate-reducing Bacteria (SRB), sulfate-reducing prokaryotes (SRP)