title: Loop-mediated isothermal amplification assay targeting the omp25 gene for rapid detection of Brucella spp.
abstract: A novel loop-mediated isothermal amplification (LAMP) assay was established to detect Brucella species DNA in milk and blood samples of animals and humans. This LAMP assay based on the sequence of highly repetitive omp25 gene was able to detect 9fg/μl Brucella spp. DNA with high sensitivity, which was 10 times higher than the nested PCR. The LAMP was evaluated for its specificity using 19 strains of six Brucella species and 28 related non-Brucella micro-organism strains as controls. The target 19 Brucella strains were all amplified, and no cross-reaction was found with all the non-Brucella micro-organism strains. Both nested PCR and LAMP assays were then used to detect Brucella spp. DNA in 78 milk samples and 113 blood samples from animals and 11 blood samples from humans, and the established LAMP assay yielded 99.0% concordance rate with the nested PCR. The LAMP assay should be a potential tool with high convenience, rapidity, sensitivity and specificity for the diagnosis of Brucellosis.
authors.affiliation: Key Laboratory of Veterinary Public Health of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, China.
authors.forename: Guo-Zhen
authors.initials: GZ
authors.lastname: Lin
authors.forename: Fu-Ying
authors.initials: FY
authors.lastname: Zheng
authors.forename: Ji-Zhang
authors.initials: JZ
authors.lastname: Zhou
authors.forename: Xiao-Wei
authors.initials: XW
authors.lastname: Gong
authors.forename: Guang-Hua
authors.initials: GH
authors.lastname: Wang
authors.forename: Xiao-An
authors.initials: XA
authors.lastname: Cao
authors.forename: Chang-Qing
authors.initials: CQ
authors.lastname: Qiu
mesh_terms: D000818:Animals; D001419:Bacteria / Q000145:classification / Q000235:genetics; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D002002:Brucella / Q000145:classification / Q000235:genetics*; D002006:Brucellosis / Q000097:blood / Q000175:diagnosis / Q000382:microbiology*; D002417:Cattle; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000097:blood / Q000235:genetics; D005260:Female; D006801:Humans; D008892:Milk / Q000382:microbiology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012756:Sheep; D013045:Species Specificity; D013696:Temperature
chemical_list: D001425:Bacterial Outer Membrane Proteins; D017931:DNA Primers; D004269:DNA, Bacterial