A Rapid Quantitative Detection Method for anaerobic denitrifying bacteria (ADNB)

摘要

There was a long detection circle and higher cost in the quantitative detection of anaerobic deitrifying bacteria (ADNB) in the wastewater of bioreactor and oil field. To solve these issues, the ADNB-MPN-PCR method, combining polymerase chain reaction (PCR) technology with most probable number, was studied as a rapid quantitative detection method for ADNB. The bacterium solution directly used for PCR amplification was prepared from wastewater, which ensured the accuracy of quantification; the universal primers, ADNB341 and ADNB870, located at 16S rDNA genes of ADNB were designed, and the amplified fragment was 548bp; the reaction system and amplification conditions of PCR were also optimized. The results indicate that the order of magnitude of this method is two magnitudes higher than that of MPN-Gries, which truly represents the actual quantity of ADNB in wastewater. The whole procedure takes only 3～4 hours and the results of the detection are extremely stable, so the cost is reduced and ADNB-MPN-PCR can be applied in practical processes.