巴尔通体巢式PCR检测方法的建立与应用  

Establishment and application of a nested PCR detection method for Bartonella

作　　者：刘宏 易山宏 孙静 杨琛琛 雷巧玲 邵建伟 LIU Hong;YI Shanhong;SUN Jing;YANG Chenchen;LEI Qiaoling;SHAO Jianwei(School of Animal Science and Technology,Foshan University,Foshan 528225,China)

机构地区：[1]佛山大学动物科技学院,广东佛山528225

出　　处：《佛山科学技术学院学报(自然科学版)》2025年第1期85-91,共7页Journal of Foshan University(Natural Science Edition)

基　　金：广州市重点研发计划农业和社会发展科技项目(202103000008)。

摘　　要：为建立一种应用于巴尔通体(Bartonella)高灵敏度、高特异性的检测方法,根据巴尔通体16S保守序列设计特异性引物,通过优化反应体系及反应条件,建立巢式PCR检测方法,并利用该方法对在2023年广州市花都区采集的100份褐家鼠脾脏样品进行检测。结果显示:所建立的巢式PCR方法特异性良好,可特异性检测出巴尔通体阳性核酸,与其他对照菌株贝氏柯克斯体、无形体、钩端螺旋体、立克次体和伯氏螺旋体阳性核酸均无交叉反应;该方法灵敏度高,最低检出限为7.66×10^(1)copies·μL^(-1);应用该方法完成100份临床样品快速检测,检出4种巴尔通体,其中1种与人类疾病有关。表明建立的巴尔通体巢式PCR检测方法特异性强、灵敏度高,可为巴尔通体的临床样品检测提供技术支持。In order to develop a highly sensitive and specific detection method for Bartonella,specific primers were designed based on the conserved 16S sequence of Bartonella.By optimizing the reaction system and conditions,a nested PCR detection method was established.This method was applied to analyze 100 spleen samples collected from Rattus norvegicus in Huadu district,Guangzhou city in 2023.The results showed that the method exhibited excellent specificity,accurately detecting without any cross-reactivity with control strains of Coxiella burnetii,Anaplasmataceae,Leptospira,Rickettsia and Borrelia.Moreover,the method displayed high sensitivity,with a minimum detection limit of 7.66×10^(1)copies·μL^(-1).Furthermore,the established method successfully detected four different Bartonella species in 100 clinical samples,including one species associated with human diseases.The nested PCR method for Bartonella detection demonstrated specificity,sensitivity,and reproducibility,providing valuable technical support for the detection of Bartonella in clinical samples.

关 键 词：巴尔通体 巢式PCR 检测方法 应用 

分 类 号：S855[农业科学—临床兽医学]