Laboratório de Genética Molecular de Micro-organismos, Departamento de Microbiologia, Instituto de Biotecnologia Aplicada à Agropecuária - BIOAGRO, Universidade Federal de Viçosa, Viçosa, Brazil.
FEMS Microbiol Lett. 2014 Mar;352(1):32-7. doi: 10.1111/1574-6968.12363. Epub 2014 Jan 6.
Bacterial respiratory diseases are responsible for considerable mortality, morbidity and economic losses in the swine industry. Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia, is one of the most important disease agents, but its identification and surveillance can be impaired by the existence of many other related bacteria in normal swine microbiota. In this work, we have evaluated a BOX-A1R-based repetitive extragenic palindromic-PCR (BOX-PCR) sequence characterised amplified region (SCAR) marker for the specific identification of A. pleuropneumoniae and its use in a multiplex PCR to detect additionally Haemophilus parasuis and Pasteurella multocida, two other major respiratory pathogens of pigs that are members of the family Pasteurellaceae. PCRs based on the BOX-SCAR fragment developed were rapid, sensitive and differentiated A. pleuropneumoniae from all swine-related members of the Pasteurellaceae family tested. Single and multiplex BOX-SCAR fragment-based PCRs can be used to identify A. pleuropneumoniae from other bacterial swine pathogens and will be useful in surveillance and epidemiological studies.
细菌性呼吸道疾病是养猪业中导致高死亡率、高发病率和高经济损失的主要原因之一。胸膜肺炎放线杆菌是引起猪传染性胸膜肺炎的病原体之一,也是最重要的病原体之一,但由于正常猪微生物群中存在许多其他相关细菌,其鉴定和监测可能会受到影响。在这项工作中,我们评估了一种基于 BOX-A1R 的重复外回文 palindromic-PCR(BOX-PCR)序列特征扩增区(SCAR)标记,用于特异性鉴定胸膜肺炎放线杆菌,并将其用于多重 PCR 检测另外两种猪的主要呼吸道病原体副猪嗜血杆菌和多杀性巴氏杆菌,它们都是巴氏杆菌科的成员。基于开发的 BOX-SCAR 片段的 PCR 快速、敏感,并将胸膜肺炎放线杆菌与测试的巴氏杆菌科所有与猪相关的成员区分开来。基于单个和多重 BOX-SCAR 片段的 PCR 可用于从其他猪病原体细菌中鉴定胸膜肺炎放线杆菌,将有助于监测和流行病学研究。