title: An assessment of whole blood and fractions by nested PCR as a DNA source for diagnosing canine ehrlichiosis and anaplasmosis.
abstract: Ehrlichiosis and anaplasmosis are tick-borne diseases. Ehrlichia canis and Anaplasma platys infect mainly white cells and platelets, respectively. The main DNA source for PCR is peripheral blood, but the potential of blood cell fractions has not been extensively investigated. This study aims at assessment of whole blood (WB) and blood fractions potential in nested PCR (nPCR) to diagnose canine ehrlichiosis and anaplasmosis. The 16S rRNA gene was amplified in 71.4, 17.8, 31.57, and 30% of the WB, granulocyte (G), mononuclear cells (M), and buffy coat (BC) samples. Compared to the WB, the sensitivity of the PCR was 42.86% for the M, and BC fractions, 21.43% for the G, and 33.33% for the blood clot (C). There was fair agreement between the WB and M, BC and C, and slight with the G. Fair agreement occurred between the nPCR and morulae in the blood smear. One animal was coinfected with A. platys and E. canis. This study provided the first evidence of A. platys infection in dogs in Paraíba, Brazil, and demonstrated that WB is a better DNA source than blood fractions to detect Ehrlichia and Anaplasma by nPCR, probably because of the plasma bacterial concentration following host cell lysis.
authors.affiliation: Departamento de Ciências Biológicas, Universidade Federal de Pernambuco, Avenue Professor Moraes Rego, s/n, Cidade Universitária, 50.670-901 Recife, PE, Brazil.
authors.forename: Tereza Emmanuelle de Farias
authors.initials: TE
authors.lastname: Rotondano
authors.forename: Alzira Maria Paiva
authors.initials: AM
authors.lastname: de Almeida
authors.forename: Elane Maria Camboim
authors.initials: EM
authors.lastname: Lustosa
authors.forename: Aline Antas
authors.initials: AA
authors.lastname: Cordeiro
authors.forename: Expedito Kennedy Alves
authors.initials: EK
authors.lastname: Camboim
authors.forename: Sérgio Santos
authors.initials: SS
authors.lastname: de Azevedo
authors.forename: Paulo Paes
authors.initials: PP
authors.lastname: de Andrade
authors.forename: Marcia Almeida
authors.initials: MA
authors.lastname: de Melo
mesh_terms: D000709:Anaplasma / Q000235:genetics / Q000302:isolation & purification*; D000712:Anaplasmosis / Q000097:blood / Q000175:diagnosis* / Q000382:microbiology; D000818:Animals; D001938:Brazil; D048429:Cell Size; D060085:Coinfection / Q000097:blood / Q000382:microbiology / Q000662:veterinary; D004269:DNA, Bacterial / Q000097:blood* / Q000235:genetics; D004283:Dog Diseases / Q000097:blood / Q000382:microbiology; D004285:Dogs; D041061:Ehrlichia canis / Q000235:genetics / Q000302:isolation & purification*; D016873:Ehrlichiosis / Q000097:blood / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D005798:Genes, Bacterial; D020459:Genes, rRNA; D006098:Granulocytes / Q000382:microbiology; D006403:Hematologic Tests / Q000379:methods; D007963:Leukocytes, Mononuclear / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000032:analysis / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S