Anaplasmosis is a disease by an intraerythrocytic rickettsial pathogen in cattle, which mainly caused by Anaplasma marginale (A. marginale). Loop-mediated isothermal amplification (LAMP) is a novel DNA amplification technology by which the target DNA can be efficiently amplified with high specificity and sensitivity under isothermal conditions. In this study, a LAMP method was developed for rapid and easy diagnosis of cattle Anaplasmosis. Three groups of specific primers targeting major surface protein 5 gene (msp5), were designed and optimized, followed by confirmation of the specificity and sensitivity of LAMP assay. The result demonstrated that A. marginale-positive sample was efficiently amplified by LAMP assay and no presence of LAMP amplicon in the four blood-borne parasites control samples, Besnoitia besnoiti, Trypanosome sp, Toxoplasma gondii and Besnoitia bigemina, was confirmed by either visual inspection or agarose gel electrophoresis. One femtogram of total genomic DNA from A. marginale-positive blood was qualified to be detected by LAMP assay, which was 100 times more sensitive than that of conventional PCR. The data demonstrated that LAMP assay was a potential of high specificity, sensitivity and time-effectiveness to diagnose Anaplasmosis in cattle herds, especially for undeveloped countries.

Key words: Anaplasma marginale, Anaplasmosis, Cattle, Loop-mediated isothermal amplification, msp5