Diagnosis of Caprine Brucellosis by Serology and Multiple PCR

Abstract

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.