title: Optimization of a molecular method for the diagnosis of canine babesiosis.
abstract: Babesiosis is a hemolytic disease caused by protozoans of the genus Babesia (Apicomplexa). This disease occurs worldwide and is transmitted by ticks to a variety of mammals, including humans. The objective of the present study was to optimize a molecular approach for the detection of a fragment of 18S rDNA of Babesia canis, Babesia vogeli, Babesia rossi or Babesia gibsoni based on a single semi-nested Polymerase Chain Reaction (PCR), and compare the efficiency of this approach with that of a simple PCR protocol. To this end, 100 blood samples collected from dogs with suspected hemoparasite infections were analyzed. A comparison of the results of simple PCR and semi-nested PCR indicated a highly significant difference (p value = 0.0000). While only five (5%) of the samples tested positive using the simple protocol, 22 (22%) were positive using the snPCR technique. The results of this study reinforce the findings of previous studies, which have demonstrated the greater sensitivity of tests based on nested or semi-nested PCR. Therefore, to avoid false-negative results due to low levels of parasitemia, we suggest the preferential use of this protocol in epidemiological studies of canine babesiosis, particularly those that require reliable estimates of the prevalence of infection.
authors.affiliation: Laboratório de Tecnologia Biomolecular, Instituto de Ciências Biológicas, Universidade Federal do Pará ? UFPA, Belém, PA, Brasil.
authors.forename: Pablo Henrique Gonçalves
authors.initials: PH
authors.lastname: Moraes
authors.affiliation: Laboratório de Tecnologia Biomolecular, Instituto de Ciências Biológicas, Universidade Federal do Pará ? UFPA, Belém, PA, Brasil.
authors.forename: Claudia Pinheiro
authors.initials: CP
authors.lastname: Rufino
authors.affiliation: Laboratório de Tecnologia Biomolecular, Instituto de Ciências Biológicas, Universidade Federal do Pará ? UFPA, Belém, PA, Brasil.
authors.forename: Thais
authors.initials: T
authors.lastname: Reis
authors.affiliation: Laboratório de Tecnologia Biomolecular, Instituto de Ciências Biológicas, Universidade Federal do Pará ? UFPA, Belém, PA, Brasil.
authors.forename: Délia Cristina Figueira
authors.initials: DC
authors.lastname: Aguiar
authors.affiliation: Instituto da Saúde e Produção Animal, Universidade Federal Rural da Amazônia ? UFRA, Belém, PA, Brasil.
authors.forename: André Marcelo Conceição
authors.initials: AM
authors.lastname: Meneses
authors.affiliation: Laboratório de Tecnologia Biomolecular, Instituto de Ciências Biológicas, Universidade Federal do Pará ? UFPA, Belém, PA, Brasil.
authors.forename: Evonnildo Costa
authors.initials: EC
authors.lastname: Gonçalves
mesh_terms: D000818:Animals; D001403:Babesia / Q000235:genetics; D001404:Babesiosis / Q000175:diagnosis*; D004275:DNA, Ribosomal / Q000032:analysis; D004283:Dog Diseases / Q000175:diagnosis* / Q000469:parasitology*; D004285:Dogs; D025202:Molecular Diagnostic Techniques / Q000592:standards; D016133:Polymerase Chain Reaction
chemical_list: D004275:DNA, Ribosomal