油田硫酸盐还原菌APS-MPN-PCR快速定量检测方法

为了解决现有油田污水中硫酸盐还原菌检测周期长、检测费用较高的问题,应用聚合酶链式反应(PCR)技术与倍比稀释法(MPN)相结合的APS-MPN-PCR法,对硫酸盐还原菌进行快速定量检测.从污水中制备了直接用于PCR扩增的菌液,保证了定量检测的准确性;建立以腺苷酰硫酸还原酶基因(APS)为靶位点的通用探针APS7F和APS8R的反应体系和扩增条件.研究结果表明,与液体稀释培养法相比,该方法检测灵敏度提高了两个数量级,操作时间缩短(整个检测过程只需要3~4 h),检测费用也降低.该方法检测结果非常稳定,真实的表征了污水中实际的SRB菌数量,可以在生产中应用.

A fast quantitative detection method of oilfield sulfate reducing bacteria

At present,the detection cycle of oilfield sulfate reducing bacteria is longer and detection cost is higher.To solve these problems,a fast quantitative detection method of oilfield sulfate reducing bacteria ——APS-MPN-PCR method—— is proposed,and which is to combine polymerase chain reaction(PCR) with most probable number(MPN).The bacterium solution for PCR amplification is directly prepared using oilfield wastewater,which ensures the accuracy of quantitative detection.And the reaction system and amplification condition are designed using universal probes APS7F and APS8R of adenosine-5'-phosphosulfate reductase in SRB.The research results show that,compared with MPN method,this method makes the detection accuracy increase two orders of magnitude.So the detection results of this method can precisely reflects the actual number of SRB in oilfield wastewater.The whole detection process only takes 3~4 hours and the detection results are extremely stable.Therefore,it can be applied in practice.