山羊布鲁氏菌病的血清学和多重PCR诊断

Raúl A. Cuevas-Jacquez, J. L. Ortega-Sánchez
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引用次数: 2

摘要

我们的目的是通过血清学和多重聚合酶链反应(PCR)来诊断山羊布鲁氏菌病。实地工作是在墨西哥杜兰戈州Mapimí市San jos de Bellavista y Bermejillo的Ejidos la Victoria Municipality otlahualilo公共土地上进行的。与生产商举行了会议,向他们解释进行这项调查的目的和可获得的利益。114只土种动物与萨宁、高山种动物杂交,颈静脉取血,其中114只在不加抗凝剂的试管中取血,用于血清检测,进行玫瑰孟加拉平板试验(RBPT); 114只全血,用于处理多重PCR。采用苯酚:氯仿:异戊醇法对产布鲁氏菌RB51和梅尔氏布鲁氏菌RM1对照菌株进行DNA提取,提取布鲁氏菌属、产布鲁氏菌、梅尔氏布鲁氏菌、猪布鲁氏菌和IS711的寡核苷酸序列。普通血清阳性率为26.31%,特拉瓦利洛市血清阳性率为41.86%,Mapimí血清阳性率为18.18%。在多重PCR样本分析中,我们发现30份样本对应于melitensis,获得了100%的灵敏度和特异性。本研究中描述的PCR技术与RBPT具有100%的灵敏度和特异性,允许在属和种之间同时鉴定,实施多重PCR变异,能够鉴定布鲁氏菌属的不同种,后者导致更好的疾病诊断。
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Diagnosis of Caprine Brucellosis by Serology and Multiple PCR
Our aim was to perform the diagnosis of caprine brucellosis by serology and multiple Polymerase Chain Reactions (PCR). The field work was conducted on the Ejidos la Victoria MunicipioTlahualilo Public Lands of San José de Bellavista y Bermejillo, Mapimí Municipality, state of Durango, Mexico. Meetings were held with the producers to explain to them the objectives and benefits to be obtained with the carrying out of this investigation. Samples were taken from 114 native breed animals crossed with Saanen and Alpine breed animals from the jugular vein, obtaining 114 blood samples in tubes without anticoagulant for the obtention of serum in order to process the Rose Bengal Plate Test (RBPT) and 114 whole blood samples for processing multiple PCR. We carried out DNA extraction of control strains of Brucella abortus RB51 and Brucella melitensis RM1 employing the phenol:chloroform:isoamyl alcohol method with the oligonucleotide sequence of Brucella genus, Brucella abortus, Brucella melitensis, Brucella suis, and IS711. General seroprevalence was 26.31% and seroprevalence by Tlahualilo Municipality was 41.86%, while this in Mapimí was 18.18%. In the multiple PCR sample analysis, we found that 30 samples corresponded to B. melitensis, obtaining 100% sensitivity and specificity. The PCR technique described in this study presented 100% sensitivity and specificity with the RBPT, allowing for the simultaneous identification, between and genus and species, the implementation of the multiple-PCR variant capable of identifying different species of the Brucella genus, the latter leading to a better diagnosis of the disease.
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