期刊
JOURNAL OF MICROBIOLOGICAL METHODS
卷 90, 期 3, 页码 292-297出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.mimet.2012.05.024
关键词
Loop-mediated isothermal amplification (LAMP); Strand displacement reaction; Rapid detection; Brucella spp.; Milk; Blood
资金
- Guangzhou Medicine and Hygiene Foundation [2006-zdi-11]
Brucella spp. are facultative intracellular bacteria that infect humans and animals. In this study, the loop-mediated isothermal amplification (LAMP) was used to detect the Brucella-specific gene omp25. Reaction conditions were optimized as temperature 65 degrees C, reaction time 60 min, Mg2+ concentration 8.0 mmol/L, polymerase content Bst DNA, 0.5 mu/L, deoxyribonucleotide concentration 1.6 mmol/L, and inner/outer primer ratio 1:8. The LAMP method was evaluated with 4 Brucella species and 29 non-Brucelia bacteria species. Positive reactions were observed on all the 4 Brucella species but not on any non-Brucella species. The limit of detection of the LAMP method was 3.81 CFU Brucella spp. Using the LAMP method, 7 of 110 raw milk samples and 5 of 59 sheep blood samples were detected positive of Brucella spp. Results indicated that LAMP is a fast, specific, sensitive, inexpensive, and suitable method for diagnosis of Brucella spp. infection. (C) 2012 Elsevier B.V. All rights reserved.
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