Development of a prmA genes quantification technique and assessment of the technique’s application potential for oil and gas reservoir exploration

Abstract

Propane-oxidizing bacteria in surface soils are often used to indicate the position of oil and gas reservoirs. As a potential replacement for the laborious traditional culture-dependent counting method, we applied real-time fluorescent quantitative polymerase chain reaction detection as a quick and accurate technology for quantification of propane-oxidizing bacteria. The propane monooxygenase gene was set as the target and the assay is based on SYBR Green I dye. The detection range was from 9.75 × 108 to 9.75 × 101 gene copies/µl, with the lowest detected concentration of 9.75 copies/µl. All coefficient of variation values of the threshold cycle in the reproducibility test were better than 1%. The technique showed good sensitivity, specificity, and reproducibility. We also quantified the propane-oxidizing bacteria in soils from three vertical 250 cm profiles collected from an oil field, a gas field, and a nonoil gas field using the established technique. The results indicated that the presence of propane monooxygenase A genes in soils can indicate an oil or gas reservoir. Therefore, this technique can satisfy the requirements for microbial exploration of oil and gas.