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Issue:ISSN 1000-7083
          CN 51-1193/Q
Director:Sichuan Association for Science and Technology
Sponsored by:Sichuan Society of Zoologists; Chengdu Giant Panda Breeding Research Foundation; Sichuan Association of Wildlife Conservation; Sichuan University
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Application of LAMP Assay of Pseudomonas aeruginosa in Microbial Control of Laboratory Mice
Author of the article:XIANG Qin, WANG Hui-juan, LIU Yang et al.
Author's Workplace:Laboratory Animal Center of Chongqing Medical University
Key Words:Pseudomonas aeruginosa; Loop-mediated isothermal amplification; mice
Abstract:In order to established a method of LAMP that detection Pseudomonas aeruginosa and application in laboratory mice microbial control. Specific primers for LAMP were designed based on Pseudomonas aeruginosa target gene sequences. Pseudomonas aeruginosa-specific LAMP method was established through optimizing the reaction systems, the temperature of LAMP is 66 degrees centigrade. The inner and loop primer concentration are 70 μmol/L and 30 μmol/L respectively. A total of 87 mice's serum samples were detected and compared with the standard detection method of Pseudomonas aeruginosa according to GB/T 14926.17-2001, positive samples were verified by PCR. We found that validation of the specificity and reproducibility of the method was performed. At the same time, the limit of LAMP detection of Pseudomonas aeruginosa was 103 times more sensitive than PCR. The detection rate of Pseudomonas aeruginosa was higher 11.5% with LAMP method in serum sample than with GB/T 14926.17-2001 method(10/87 vs. 0/87). Above all, we demonstrated that LAMP method has the advantages of specificity repeatability and stability in the detection of Pseudomonas aeruginosa in mice, providing a method for the detection of P. aeruginosa.
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