title: A rapid, simple and sensitive loop-mediated isothermal amplification method to detect Anaplasma bovis in sheep and goats samples.
abstract: A loop-mediated isothermal amplification (LAMP) technique has been widely used in detecting the nucleic acid of various pathogenic bacteria. In this study, a set of four LAMP primers was designed to specifically test Anaplasma bovis. The LAMP assay was performed at 62°C for 60min in a water bath. The specificity was confirmed by amplifying A. bovis isolate, while no cross reaction was observed with other five pathogens (Anaplasma bovis, Anaplasma phagocytophilum, Theileria luwenshuni, Babesia motasi and Schistosoma japonicum). The sensitivity of LAMP was 5×100copies/μL, 100 times more than that of conventional PCR (5×102copies/μL). Of 120 blood DNA extracted from sheep and goats field samples, 81 (67.5%), 22 (18.3%) and 43 (35.8%) were positively detected by LAMP, conventional PCR and nested PCR, respectively. The findings indicated that the developed LAMP assay is a new convenient tool for rapid and cost-effective detection of A. bovis.
authors.affiliation: Parasitology Laboratory, College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, PR China.
authors.forename: Jinhong
authors.initials: J
authors.lastname: Wang
authors.affiliation: Parasitology Laboratory, College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, PR China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Parasitology Laboratory, College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, PR China.
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Cui
authors.affiliation: Parasitology Laboratory, College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, PR China.
authors.forename: Yaqun
authors.initials: Y
authors.lastname: Yan
authors.affiliation: Parasitology Laboratory, College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, PR China.
authors.forename: Xiaoxing
authors.initials: X
authors.lastname: Wang
authors.affiliation: Parasitology Laboratory, College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, PR China.
authors.forename: Rongjun
authors.initials: R
authors.lastname: Wang
authors.affiliation: Parasitology Laboratory, College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, PR China.
authors.forename: Fuchun
authors.initials: F
authors.lastname: Jian
authors.affiliation: Parasitology Laboratory, College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, PR China.
authors.forename: Longxian
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Parasitology Laboratory, College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, PR China. Electronic address: nnl1986@163.com.
authors.forename: Changshen
authors.initials: C
authors.lastname: Ning
mesh_terms: D000709:Anaplasma / Q000302:isolation & purification*; D000712:Anaplasmosis / Q000175:diagnosis* / Q000469:parasitology; D000818:Animals; D016054:DNA, Protozoan / Q000235:genetics; D015511:Goat Diseases / Q000175:diagnosis* / Q000469:parasitology; D006041:Goats; D021141:Nucleic Acid Amplification Techniques / Q000191:economics / Q000662:veterinary*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis* / Q000469:parasitology
chemical_list: D016054:DNA, Protozoan; D012336:RNA, Ribosomal, 16S
keywords: 16S rRNA; Anaplasma bovis; LAMP assay