机构：[1]Molecular Diagnosis Laboratory, Faculty of Life Science and Technology, Kunming University of Science and Technology[2]Molecular Medicine Center of Yunnan Province, Kunming, Yunnan 650500[3]Department of Clinical Laboratory, First People's Hospital of Yunnan Province, Kunming, Yunnan 650034, P.R. China医技片检验科云南省第一人民医院
Pseudomonas aeruginosa is one of the three most pathogenic bacteria that frequently cause life-threatening opportunistic human infections, pneumonia, and lower respiratory tract infections in immunocompromised hosts, particularly in the burns ward. The present study aimed to establish a loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of P. aeruginosa-specific gene hypothetical protein (GenBank ID: 882161). The gene was obtained through local and online BLAST, and specific primers were designed for this gene. Reaction conditions were optimized at 65 degrees C for 30 min and 80 degrees C for 2 min, whereas the reaction system contained 5.2 mM Mg2+, 8 U Bst 2.0 DNA polymerase, 1.4 mM deoxyribonucleotide and 0.2 and 1.6 mu M of the outer and inner primers, respectively. The LAMP method was evaluated using 150 P. aeruginosa and 170 non-P. aeruginosa strains. Positive reactions were observed on 150 P. aeruginosa strains, whereas all non-P. aeruginosa strains exhibited negative results. Plasmids with the specific gene and mouse blood with P. aeruginosa were used for sensitivity assay. The detection limit of LAMP was 1 bacterium/reaction. Results indicated that the LAMP method targeted to hypothetical protein is a fast, specific, sensitive, inexpensive and suitable method for detection of P. aeruginosa.
基金：
Yunnan Science and Technology Commission [2015BC001, 2015DH010]
第一作者机构：[1]Molecular Diagnosis Laboratory, Faculty of Life Science and Technology, Kunming University of Science and Technology[2]Molecular Medicine Center of Yunnan Province, Kunming, Yunnan 650500
通讯作者：
通讯机构：[1]Molecular Diagnosis Laboratory, Faculty of Life Science and Technology, Kunming University of Science and Technology[2]Molecular Medicine Center of Yunnan Province, Kunming, Yunnan 650500[*1]Molecular Diagnosis Laboratory, Faculty of Life Science and Technology, Kunming University of Science and Technology, 727 Jingming South Road, Kunming, Yunnan 650500, P.R. China
推荐引用方式(GB/T 7714)：
Li Chao,Shi Yaoqiang,Yang Guangying,et al.Establishment of loop-mediated isothermal amplification for rapid detection of Pseudomonas aeruginosa[J].EXPERIMENTAL AND THERAPEUTIC MEDICINE.2019,17(1):131-136.doi:10.3892/etm.2018.6910.
APA：
Li, Chao,Shi, Yaoqiang,Yang, Guangying,Xia, Xue Shan,Mao, Xiaoqin...&Song, Yuzhu.(2019).Establishment of loop-mediated isothermal amplification for rapid detection of Pseudomonas aeruginosa.EXPERIMENTAL AND THERAPEUTIC MEDICINE,17,(1)
MLA：
Li, Chao,et al."Establishment of loop-mediated isothermal amplification for rapid detection of Pseudomonas aeruginosa".EXPERIMENTAL AND THERAPEUTIC MEDICINE 17..1(2019):131-136
