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. marginale DNA 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 msp1α gene was developed for detection and quantification of A. marginale in bovine blood. The real-time PCR assay is able to detect as few as one copу of msp1α gene 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. marginale in infected cattle, and for the evaluation of the efficacy of anti-rickettsial drugs and anaplasmosis vaccines.

牛无形体病是由tick传播的立克次体病，在许多国家造成重大的经济损失。牛无形体病的主要病原是边缘无形体（Rickettsiales，Anaplasmataceae）。迄今为止，已经提出了几种PCR检测方法，用于边缘A曲霉DNA检测，但是它们中的大多数不提供内部扩增对照，这可以防止假阴性结果，并且对于通过PCR检测病原体DNA检测结果的可靠性是必需的。 。在本研究中，开发了一种基于msp1α基因的物种特异性且高度保守的片段的实时PCR检测试剂盒，用于检测和定量分析margin缘拟南芥。在牛血中。实时荧光定量PCR检测每个反应最多可检测到多达msp1α基因。为防止假阴性结果，提供了同时扩增和检测牛基因组DNA片段作为内源性内部扩增对照（IAC）的功能。所述测定法可以被用作用于检测和定量一种高度特异和灵敏的方法A.边缘无在感染牛，以及用于抗立克次体药和边虫病疫苗的效力的评估。