油田硫酸盐还原菌快速定量检测方法

现有油田废水中硫酸盐还原菌检测周期长、检测费用较高,本研究应用聚合酶链式反应(PCR)技术与倍比稀释法(MPN)相结合的DSR-MPN-PCR法,对硫酸盐还原菌进行快速定量检测.从废水中制备了直接用于PCR扩增的菌液,保证了定量准确性;建立以硫酸盐还原菌亚硫酸盐还原酶基因(Dsr)为靶位点的通用探针DSR1F和DSR5R的反应体系和扩增条件.结果表明,该方法检测灵敏度明显比液体稀释培养法高2个数量级,真实地表征了废水中实际的SRB菌数量,整个操作过程需要3～4　h,检测结果非常稳定,降低了检测费用,可以在生产中应用.

Rapid Quantitative Detection of Sulfate Reducing Bacteria in Oilfield

It take long time and high cost to measure sulfate reducing bacteria (SRB) in wastewater of oil field. A rapid quantitative method was developped by combining polymerase chain reaction(PCR) and most probable number(MPN) to measure sulfate reducing bacteria (SRB) in wastewater of oil field. The bacterium solution was directly prepared from wastewater for PCR amplification, which ensured quantitative accuracy. Reaction system and amplification condition were designed using universal primers DSR1F and DSR5R of dissimilatory sulfite reductase in SRB. The result showed that the accuracy of this method is two magnitude higher than that of MPN. The whole measuring process taked 3~4 hours and the reproducibility of this method is extremely stable, being fit to practical process.