城市生活污水中志贺氏菌的定量PCR检测研究-市政工程专业论文.docxVIP

The research of Quantitative PCR Detection of Shigella in Municipal sewage Speciality: Municipal engineering Author:Sun Tingting Supervisor: Prof. Wang Xiaochang Abstract Water pollution is one of the environment problem China facing seriously. In order to ease water shortage, we do more and more work to control water pollution, and the water quality of sewage after secondary treatment has been improved, but the absolute number 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 epithelium cells, causing diarrhea, abdominal pain, fever, et.al. and doing harm to human health. In view 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 the improvement of the sewage treatment process for removing pathogens and the evaluation of the risks of pathogens. This research selected membrane adsorption-elution as sample pre-treatment method for sewage water sample. We detected the concentrated Shigella using the selected primers ipaH8 and ipaH10. Through gradient PCR,we analyse the influence of the annealing temperature to the results of detection,and at last we choose 57 ℃ as the annealing temperature. The quantitative PCR method to detect the ipaH gene of Shigella was established on based of the qualitative detection method. Combining the distribution of Shigella in xi`an ,we solve the problem caused by the ipaH gene multiple-copy nature. In addition, we simulated the natural water, and researched the changing patterns of Shigella using the real-time polymerase chain reaction established above. At the same time, water samples from A and B two different of wastewater treatment plan in xian area was detected, and an analysis of the distrbution of Shigella present in secondary effluent was made. Key words:ipaH gene; Shigella;real-time polymerase chain reaction; multipl