恙虫病东方体与莫氏立克次体双重实时荧光PCR检测方法建立及其在鼠类检测中的应用

Establishment of a Dual Real-Time Fluorescent PCR Detection Method for Detection of Orientiatsutsugamushi and Rickettsia Mooseriand Its Application in the Detection of Rodents

目的建立一种基于TaqMan-MGB探针技术的双重实时荧光PCR检测方法,用于鼠类携带恙虫病东方体和莫氏立克次体的检测工作开展。方法依据恙虫病东方体56-kDa蛋白基因序列设计恙虫病东方体引物及Taqman-MGB探针,参照文献合成莫氏立克次体引物及Taqman-MGB探针,建立恙虫病东方体及莫氏立克次体双重实时荧光PCR检测方法。结果利用本文建立的双重实时荧光PCR检测方法对288份鼠肺脏组织样品进行检测,检测结果与普通PCR检测结果一致。结论本文建立的双重实时荧光PCR检测方法,具有较好的特异性、敏感性及重复性,满足国境口岸鼠类携带恙虫病东方体及莫氏立克次体的监测检测工作需要。

Objective To Establish a dual real-time fluorescent PCR detection method based on a TaqMan-MGB probe technology for the detection of orientiatsutsugamushi and rickettsia moosericarried by rodents. Methods Based on 56 - kDa protein gene sequence,we designed orientiatsutsugamushi primers and Taqman-MGB probe and reference literature tosynthetic rickettsiamooseri primers and Taqman-MGB probe to establish a dual real-time fluorescent PCR detection method. Results A total of 288 rat lung tissue samples were detected by the dual-real-time fluorescence PCR method,and the results were consistent with the results of ordinary PCR. Conclusion The double real-time fluorescent PCR method established in this paper has good specificity,sensitivity and repeatability,whichcan meet that requirement of the monitor and detection of theorientiatsutsugamushiand rickettsia mooseriatthe frontier port.