| Abstrakt: |
Bovine anaplasmosis is a tick-borne rickettsial disease, causing significant economic losses in many countries. The main causative agent of bovine anaplasmosis is Anaplasma marginale(Rickettsiales, Anaplasmataceae). To date, several PCR assays for A. marginaleDNA detection were proposed, but most of them do not provide an internal amplification control, which allows to prevent false-negative results and is required for reliability of the results of pathogen DNA detection by PCR assay. In the present study, a real-time PCR assay based on the species-specific and highly conserved fragment of msp1αgene was developed for detection and quantification of A. marginalein bovine blood. The real-time PCR assay is able to detect as few as one copу of msp1αgene per reaction. To prevent false-negative results, simultaneous amplification and detection of the bovine genomic DNA fragment as an endogenous internal amplification control (IAC) was provided. The assay can be used as a highly specific and sensitive method for detection and quantification of A. marginalein infected cattle, and for the evaluation of the efficacy of anti-rickettsial drugs and anaplasmosis vaccines. |
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