食品工业科技2020，Vol.41Issue(11)：267-272,293,7.DOI:10.13386/j.issn1002-0306.2020.11.041
铜绿假单胞菌LAMP快速检测方法建立及微滴数字PCR验证
Establishment of LAMP Rapid Detection Method for Pseudomonas aeruginosa and Verification of Droplet Digital PCR
摘要
Abstract
Objective: A rapid method for detecting Pseudomonas aeruginosa was establish by loop-mediated isothermal amplification, and verified by microdroplet digital PCP assay. Methods: Using the rpod gene of Pseudomonas aeruginosa as the target gene, a rapid detection method for Pseudomonas aeruginosa in juice drink was established by design of primers. For the reliability, specificity and senisitivity of the method was verified by the droplet digital PCP. Results: The established LAMP rapid detection method had obviously color change. In the detection of droplets digital PCR, the concentration of Pseudomonas aeruginosa suspension was used as abscissa, and the number of copy amplified by digital PCR was used as ordinate. The standard curve was y=0.07497 x-9.3516.The linear relationship was good,R2=0.9999.The detection range of Pseudomonas aeruginosa was 1.5 × 102~1.5 × 105 CFU/mL The method had good specificity, high sensitivity and accuracy. The detection limit of standard Pseudomonas aeruginosa was 1 ng/μL. The standard deviation were 0.57，0.71，4.24，14.8，and the RSD value between 0.66% to 22.8% . 128 samples of juice drinks were detected, 126 samples were not colored, and 2 samples were colored.The valves were determined by ddPCR method, and the concentrations were 1.64 × 104 and 2.29 × 102 CFU/mL, respectively.关键词
铜绿假单胞菌/环介导等温扩增/快速检测/微滴数字PCRKey words
Pseudomonas aeruginosa/loop-mediated isothermal amplification/rapid detection/droplet digital PCR分类
轻工纺织引用本文复制引用
李羽翡1,祖新1,李羽翠2,张德荣3*..铜绿假单胞菌LAMP快速检测方法建立及微滴数字PCR验证[J].食品工业科技,2020,41(11):267-272,293,7.基金项目
甘肃省食品药品重点研发项目(2018GSFDA038). （2018GSFDA038）
