Evaluation of Propidium Monoazide as a Tool to Differentiate Live from Dead MIC Microorganisms

摘要

We describe the advancement of an activity-based qPCR assay which can distinguish live from dead corrosion influencing microorganisms in oil and gas pipeline environments. While the current body of knowledge is expanding for the types of microbial organisms present in waters used for natural gas and oil production, there is scant information on the activity of these potentially deleterious or beneficial organisms in source waters and produced waters associated with the industry. This assay was developed using pure cultures of sulfate reducing bacteria (SRB), sulfate reducing Archaea (SRA), iron oxidizing bacteria (IOB), denitrifying bacteria (DNB), and archaea (AR). Propidium monoazide-qPCR was then adapted to differentiate live from dead bacteria Pseudomonas putida bacterial cells once immobilized on membrane filters. The assay has been validated with mock environmental samples, where produced waters of various turbidity and origin have been sterilized and spiked with known quantities of living and dead P.putida. We discuss the limitations and possible future optimization methods for Propidium monazide-qPCR techniques in the industry.