The Research Of Quantitative PCR Detection Of Shigella In Municipal Sewage

Water pollution is one of the environment problem China facing seriously. In orderto ease water shortage, we do more and more work to control water pollution, and thewater quality of sewage after secondary treatment has been improved, but the absolutenumber of bacteria in the sewage is still considerable, so it does harm to human health.Shigella is one of the typical pathogenic bacteria, it invades human intestinal epitheliumcells, causing diarrhea, abdominal pain, fever, et.al. and doing harm to human health. Inview of this, the detection of Shigella in urban wastewater reclamation and reuse,through which the presence of it can be known, is practically significant to theimprovement of the sewage treatment process for removing pathogens and theevaluation of the risks of pathogens.This research selected membrane adsorption-elution as sample pre-treatmentmethod for sewage water sample. We detected the concentrated Shigella using theselected primers ipaH8and ipaH10. Through gradient PCR,we analyse the influence ofthe annealing temperature to the results of detection,and at last we choose57℃astheannealing temperature. The quantitative PCR method to detect the ipaH gene of Shigellawas established on based of the qualitative detection method. Combining thedistribution of Shigella in xi`an,we solve the problem caused by the ipaH genemultiple-copy nature. In addition, we simulated the natural water, and researched thechanging patterns of Shigella using the real-time polymerase chain reaction establishedabove. At the same time, water samples from A and B two different of wastewatertreatment plan in xian area was detected, and an analysis of the distrbution of Shigella present in secondary effluent was made.