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目的建立城市污水二级处理出水中伤寒沙门菌的定量PCR检测方法。方法以构建的伤寒沙门菌重组质粒作为标准品,采用whatman2号(孔径为8μm)定性滤纸对水样预过滤,硝酸纤维素膜浓缩水样,选用伤寒沙门菌特异性引物ST3和ST4进行实时荧光定量PCR(quantitative PCR,QPCR)检测。结果在2.8×101～2.8×108基因组相当量拷贝数(genome equivalent copy,GEC)模板量的线性范围内,所得回归方程线性关系较好(r=0.995 5)。该方法的检测灵敏度为1.4×101 GEC/μl。结论该方法快速、特异、灵敏,适用于城市污水厂二级处理出水的伤寒沙门菌的定量检测。
Objective To establish a quantitative PCR method for the detection of Salmonella typhi in secondary effluent from municipal wastewater treatment. Methods The constructed Salmonella typhi recombinant plasmids were used as standard samples. The samples of whatman number 2 (pore size 8μm) were used to pre-filter the water samples and the nitrocellulose membrane was used to concentrate the water samples. Salmonella typhimurium specific primers ST3 and ST4 were used for real-time fluorescence Quantitative PCR (QPCR) detection. The results showed that the regression equation was linear within the linear range of the amount of genome equivalent copy (GEC) template from 2.8 × 101 to 2.8 × 108 (r = 0.995 5). The detection sensitivity of this method is 1.4 × 101 GEC / μl. Conclusion The method is rapid, specific and sensitive and is suitable for the quantitative detection of Salmonella typhi isolated from secondary effluent of urban sewage treatment plants.