title: Loop-mediated isothermal amplification (LAMP) method based on two species-specific primer sets for the rapid identification of Chinese Babesia bovis and B. bigemina.
abstract: Bovine babesiosis is a tick-transmitted hemoprotozoan disease that is mainly caused by Babesia bovis and/or Babesia bigemina and is characterized by significant morbidity and mortality worldwide. This disease is widespread in most parts of China. However, it is difficult to rapidly discriminate between the B. bovis and B. bigemina species. To detect and distinguish these species, a loop-mediated isothermal amplification (LAMP) platform that targets specific sequences of the internal transcribed spacer (ITS) genes was developed. Specificity testing revealed that there was no cross-reaction with the other tick-borne parasites B. ovate, B. major, unnamed bovine Babesia, Theileria annulata, Theileria sinensis, Theileria sergenti, and Anaplasma marginale, or with bovine white blood cells. The sensitivity of the LAMP method was 0.1 pg DNA for both B. bovis and B. bigemina, which was superior to that of the classical PCR methods. This assay was evaluated for its diagnostic utility using blood samples collected from experimentally and naturally infected cattle in China. These findings indicate that the Babesia species-specific LAMP assay may have potential clinical application in the detection and differentiation of Babesia species, particularly in countries in which babesiosis is endemic.
authors: 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: Aihong
authors: A
authors: Liu
authors: Guiquan
authors: G
authors: Guan
authors: Pengfei
authors: P
authors: Du
authors: Huitian
authors: H
authors: Gou
authors: Zhijie
authors: Z
authors: Liu
authors: Junlong
authors: J
authors: Liu
authors: Milin
authors: M
authors: Ma
authors: Jifei
authors: J
authors: Yang
authors: Youquan
authors: Y
authors: Li
authors: Qinli
authors: Q
authors: Niu
authors: Qiaoyun
authors: Q
authors: Ren
authors: Qi
authors: Q
authors: Bai
authors: Hong
authors: H
authors: Yin
authors: Jianxun
authors: J
authors: Luo
mesh_terms: D000818:Animals; D001403:Babesia / Q000145:classification* / Q000235:genetics*; D001404:Babesiosis / Q000453:epidemiology / Q000469:parasitology / Q000662:veterinary*; D002417:Cattle; D002681:China / Q000453:epidemiology; D021901:DNA, Intergenic / Q000235:genetics; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D021901:DNA, Intergenic