猪鼻腔棉拭子中胸膜肺炎放线杆菌检测与分离方法的优化  

Optimization of the detection and isolation method for Actinobacillus pleuropneumonia in swine nasal swab sample

作　　者：孙裴[1] 施雷[1] 魏建忠[1] 耿照玉[1] 殷宗俊[1] 顾秋成[1] 王海洋[1] 李好磊 郭志英[1] 李郁[1] 

机构地区：[1]安徽农业大学动物科技学院,合肥230036

出　　处：《安徽农业大学学报》2015年第5期706-710,共5页Journal of Anhui Agricultural University

基　　金：安徽省畜禽产业共性技术研究院;现代农业产业技术体系建设专项资金(CARS-36-生猪);安徽省生猪产业技术体系资金项目共同资助

摘　　要：为提高APP亚临床感染的核酸检出率和细菌分离率,通过检测3种PCR的敏感性和特异性,运用3种PCR方法分别检测母猪鼻腔棉拭子中APP核酸;并同时使用4种选择性培养基分离母猪鼻腔棉拭子中APP。结果显示,3种PCR的最低检测限度均为102 CFU,且除APP外与其他细菌不发生反应;dsb E-PCR方法的App核酸检出率(23.7%)显著高于oml A-PCR(8.1%)和Apx IVA-PCR(14.8%),三者符合率较高,分别为84.4%(dsb E/oml A-PCR)、91.1%(dsb E/Apx IVA-PCR)和93.3%(oml A/Apx IVA-PCR);m PPLO的APP分离率(11.9%)显著高于TSA(0.7%)、CA(5.2%)和BA(2.2%),四者符合率较高,分别为88.9%(m PPLO/TSA)、93.3%(m PPLO/CA)、90.37%(m PPLO/BA)、95.6%(TSA/CA)、98.5%(TSA/BA)和97.0%(CA/BA);dsb E-PCR检测结果与m PPLO分离结果的符合率较高(88.1%)。说明dsb E-PCR和m PPLO的检测分离效率最高,可综合2种方法,完善APP检测分离操作程序,为APP亚临床感染的诊断和治疗以及疫苗免疫提供科学依据。In order to improve the nucleic acid detection rate and bacterial isolation rate of APP subclinical infection, the sensitivity and specificity of three PCR methods used to detect APP nucleic acid nasal swab in sows were evaluated. The result of bacteria isolation using four selective media was also investigated. The results showed that the minimum detection limit using three PCR methods was 102 CFU, and it had no nonspecific amplification with other bacteria. The nucleic acid detection rate of dsb E-PCR method（23.7%） was significantly higher than that of oml A-PCR（8.1%） and Apx IVA-PCR（14.8%）, and the coincidence rates were 84.4%（dsb E/oml A-PCR）, 91.1%（dsb E/Apx IVA-PCR） 93.3%（oml A/Apx IVA-PCR）. The APP isolation rate of m PPLO（11.9%） was significantly higher than that of TSA（0.7%）, CA（5.2%） and BA（2.2%）, and the coincidence rates were 88.9%（m PPLO/TSA）, 93.3%（m PPLO/CA）, 90.37%（m PPLO/BA）, 95.6%（TSA/CA）, 98.5%（TSA/BA） and 97.0%（CA/BA）. The coincidence rates of the detection results of dsb E-PCR and m PPLO were relatively high（88.1%）. It indicated that Dsb EPCR and m PPLO methods have high separation efficiency, which can be used together to improve APP detection and separation procedures, and provide scientific basis for diagnosis and treatment of subclinical infection of APP.

关 键 词：胸膜肺炎放线杆菌 PCR 选择性培养基 优化 

分 类 号：S855.1[农业科学—临床兽医学]