实时荧光定量PCR法检测环境假单胞菌属细菌丰度

Detection of abundance of Pseudomonas in environmental samples by real-time quantitative PCR

应用实时荧光定量PCR法，建立了总细菌及假单胞菌属的标准曲线，并以环境样本中假单胞菌属与总细菌的比值反映其细菌丰度．应用该方法评价人工除藻反应器中组合填料、无纺布、弹性填料等介质对太湖水中溶藻细菌之一的假单胞菌属的富集情况．结果显示：该方法对总细菌的检测范围为10^3～10^8个基因拷贝/μL（R^2=0．997）；假单胞菌属的检测范围为1～10^5个基因拷贝／μL（R^2=0．994）．对除藻反应器溶藻细菌富集效果的评价显示，所建立的方法能有效检测反应器中填料对该类溶藻细菌的富集程度．初步证明所建立的方法可有效定量假单胞菌属在环境样本中的丰度．

A real-time quantitative PCR （polymerase chain reaction） （RTQ-PCR） method is established to quantitatively detect the abundance of algicidal bacteria of Pseudomonas in environmental samples in this study. The standard abundance curves of total bacteria and Pseudomonas were established respectively and the abundance of Pseudomonas was demonstrated by the Ct ratio of Pseudomonas and total bacteria. The abundance of algicidal bacteria of Pseudomonas on the artificial mediums of algae treatment reactor in Taihu Lake were then detected by the method. The results show that the determination ranges of total bacteria and Pseudomonas are 10^3 -10^8gene copies/μL （R^2 = 0. 997 ） and 1 - 10^5 gene copies/μL （R^2 =0. 994）, respectively. The accumulation of Pseudomonas on the artificial mediums in reactor can be found effectively by the assay. It is believed that RTQ-PCR method can detect the abundance of algicidal bacteria of Pseudomonas in environmental samples quantitatively.