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Your Position :Home->Past Journals Catalog->2019 Vol.38 No.1

Application of Loop-Mediated Isothermal Amplification Assay of Pseudomonas aeruginosa in Microbial Control of Laboratory Mice
Author of the article:XIANG Qin1, WANG Huijuan1,2, LIU Yang1, WU Yue3, CAO Min1, LAI Guoqi1, HE Mingzhong1*
Author's Workplace:1. Laboratory Animal Center of Chongqing Medical University, Chongqing 400016, China;
2. School of Medicine, Zhejiang University, Hangzhou 310058, China;
3. Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Chongqing Medical University, Chongqing 400010, China
Key Words:Pseudomonas aeruginosa; loop-mediated isothermal amplification; mice
Abstract:In order to established an assay of loop-mediated isothermal amplification (LAMP) for Pseudomonas aeruginosa detection in laboratory mice. Specific primers for LAMP were designed based on P. aeruginosa oprL gene. P. aeruginosa-specific LAMP assay was established through optimizing the reaction system, and the optimal temperature of LAMP was found to be 66 ℃. The inner and loop primer concentration were 70 μmol·L-1 and 30 μmol·L-1, respectively. A total of 87 mouse serum samples were detected and compared with the standard assay of P. aeruginosa according to GB/T 14926.17-2001, and the P. aeruginosa-positive samples were further verified by PCR. Validation of the specificity and reproducibility of the LAMP assay was also performed. Finally, the limit of LAMP detection for P. aeruginosa was 103 times more sensitive than normal PCR assay. The detection rate of P. aeruginosa by LAMP assay in serum samples was higher than that by GB/T 14926.17-2001 assay (10/87 vs. 0/87). Therefore, we demonstrate that LAMP assay has the advantages of specificity, sensitivity, repeatability, and stability in detecting P. aeruginosa in mice.
2019,38(1): 28-36 收稿日期:2018-06-21
分类号:Q95-33
基金项目:国家自然科学基金面上项目(81570541);重庆市科委民生项目(cstc2017shmsA130097)
作者简介:向钦(1991—),女,硕士研究生,实验动物学,E-mail:xqin0512@163.com
*通信作者:何明忠,E-mail:564290461@qq.com
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