title: Rapid identification and differentiation of Theileria sergenti and Theileria sinensis using a loop-mediated isothermal amplification (LAMP) assay.
abstract: The present study developed and validated a species-specific loop-mediated isothermal amplification (LAMP) assay for the rapid detection and discrimination of two benign bovine Theileria species -T. sergenti and T. sinensis. The LAMP assay is inexpensive and easy to perform and involves a rapid reaction-the amplification can be performed in 55 min or 50 min under isothermal conditions of 61°C or 63°C, respectively, by employing a set of four species-specific primer mixtures. The results can be checked using agarose gels. The optimal assay conditions, under which the assay exhibited with no cross-reaction with other closely related tick-borne parasites (T. annulata, Babesia bovis, B. bigemina, B. major, B. ovata, B. U. sp., Anaplasma marginale) or between the two Theileria species of interest, was established. The assay is approximately 10-fold more sensitive than the conventional specific PCR assay. The LAMP assay was validated using DNA from 6 standard stocks in the laboratory and was evaluated for its diagnostic utility using blood samples collected from experimentally and naturally infection cattle or yaks in China. These findings indicate that this Theileria species-specific LAMP assay may have potential clinical applications for the detection and differentiation of two benign bovine Theileria species -T. sergenti and T. sinensis, especially in endemic countries.
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou, Gansu 730046, PR China.
authors.forename: Aihong
authors.initials: A
authors.lastname: Liu
authors.forename: Guiquan
authors.initials: G
authors.lastname: Guan
authors.forename: Pengfei
authors.initials: P
authors.lastname: Du
authors.forename: Huitian
authors.initials: H
authors.lastname: Gou
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.forename: Zhijie
authors.initials: Z
authors.lastname: Liu
authors.forename: Milin
authors.initials: M
authors.lastname: Ma
authors.forename: Qiaoyun
authors.initials: Q
authors.lastname: Ren
authors.forename: Junlong
authors.initials: J
authors.lastname: Liu
authors.forename: Jifei
authors.initials: J
authors.lastname: Yang
authors.forename: Youquan
authors.initials: Y
authors.lastname: Li
authors.forename: Qinli
authors.initials: Q
authors.lastname: Niu
authors.forename: Qi
authors.initials: Q
authors.lastname: Bai
authors.forename: Hong
authors.initials: H
authors.lastname: Yin
authors.forename: Jianxun
authors.initials: J
authors.lastname: Luo
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000469:parasitology*; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000592:standards*; D010274:Parasitology / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013045:Species Specificity; D016794:Theileria / Q000235:genetics / Q000502:physiology*; D013801:Theileriasis / Q000175:diagnosis / Q000469:parasitology*
chemical_list: D021903:DNA, Ribosomal Spacer