title: Loop-mediated isothermal amplification for the rapid detection of Haemophilus parasuis.
abstract: Haemophilus parasuis infection is of considerable economic importance in the swine industry due to the high costs associated with treatment and loss of animals all over the world. In the present study, loop-mediated isothermal amplification (LAMP) is described for the rapid and specific detection of this species. A primer set derived from the inf B gene of H. parasuis was used to validate the assay using 15 H. parasuis reference strains, 39 clinical isolates, 75 positive samples, and 18 other pathogens. The results indicated that positive reactions were confirmed for all H. parasuis strains and specimens by LAMP after 45 min reaction at 65 °C in a water bath, and no cross-reactivity was observed from other non-H. parasuis strains. The detection limit of the conventional PCR was 25 copies, while that of the LAMP was five copies per tube. Therefore, the sensitivity of LAMP was higher than that of PCR. LAMP is likely to be more suitable as a routine diagnostic tool, especially in clinics without complicated equipment such as thermal cycling machines and electrophoresis apparatus. In these scenarios, the H. parasuis LAMP assay has the potential for field diagnosis.
authors.affiliation: Key Laboratory of Animal Virology of Ministry of Agriculture, Key Laboratory of Grazing Animal Disease of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.
authors.forename: Hao-Tai
authors.initials: HT
authors.lastname: Chen
authors.forename: Yue-Feng
authors.initials: YF
authors.lastname: Chu
authors.forename: Yong-Sheng
authors.initials: YS
authors.lastname: Liu
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Zhong-Xin
authors.initials: ZX
authors.lastname: Lu
mesh_terms: D000818:Animals; D001431:Bacteriological Techniques / Q000379:methods*; D003429:Cross Reactions; D017931:DNA Primers / Q000235:genetics; D006192:Haemophilus Infections / Q000175:diagnosis / Q000662:veterinary*; D044137:Haemophilus parasuis / Q000235:genetics / Q000302:isolation & purification*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D039881:Prokaryotic Initiation Factor-2 / Q000235:genetics; D012680:Sensitivity and Specificity; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000382:microbiology; D013696:Temperature
chemical_list: D017931:DNA Primers; D039881:Prokaryotic Initiation Factor-2