title: Molecular and serological detection of A. centrale- and A. marginale-infected cattle grazing within an endemic area.
abstract: A reverse line blot hybridization (RLB) one-stage nested PCR (nPCR) for Anaplasma centrale and a nested PCR for Anaplasma marginale were used to detect infected cattle grazing within an endemic region in Israel. A novel set of PCR primers and oligonucleotide probes based on a 16S ribosomal RNA gene was designed for RLB detection of both Anaplasma species, and the performance of the molecular assays compared. The immunofluorescent antibody test (IFA) was used to detect antibodies to both Anaplasma species, whereas, a highly sensitive and specific competitive enzyme-linked immunosorbent assay (cELISA) was used to detect antibodies in A. centrale-vaccinated cattle. The RLB and the nested PCR procedures showed bacteremia with sensitivity of 50 infected erythrocytes per milliliter. Up to 93% of the A. centrale vaccinates carried specific antibodies that were detected by cELISA, and up to 71% of the vaccinated cattle were found to be naturally infected with A. marginale according to the PCR and the RLB assays. Nevertheless, no severe outbreaks of A. marginale infection occurred among vaccinated herds in this endemic region. It appears that both, molecular tools and serology are useful for evaluation of the vaccine efficacy. In the light of wide natural field infection with A. marginale, strong recommendations to continue the A. centrale vaccination program regime will continue until a new generation of non-blood-based vaccine will be developed.
authors.affiliation: Division of Parasitology, Kimron Veterinary Institute, P.O. Box 12, Bet Dagan 50250, Israel.
authors.forename: T
authors.initials: T
authors.lastname: Molad
authors.forename: M L
authors.initials: ML
authors.lastname: Mazuz
authors.forename: L
authors.initials: L
authors.lastname: Fleiderovitz
authors.forename: L
authors.initials: L
authors.lastname: Fish
authors.forename: I
authors.initials: I
authors.lastname: Savitsky
authors.forename: Y
authors.initials: Y
authors.lastname: Krigel
authors.forename: B
authors.initials: B
authors.lastname: Leibovitz
authors.forename: J
authors.initials: J
authors.lastname: Molloy
authors.forename: F
authors.initials: F
authors.lastname: Jongejan
authors.forename: V
authors.initials: V
authors.lastname: Shkap
mesh_terms: D042321:Anaplasma centrale / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D042322:Anaplasma marginale / Q000235:genetics / Q000302:isolation & purification*; D000712:Anaplasmosis / Q000175:diagnosis* / Q000382:microbiology / Q000517:prevention & control; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D001428:Bacterial Vaccines / Q000276:immunology; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000382:microbiology / Q000517:prevention & control; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry; D004269:DNA, Bacterial / Q000737:chemistry; D019353:Endemic Diseases / Q000662:veterinary; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D007557:Israel; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity
chemical_list: D000907:Antibodies, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial; C450434:major surface protein 1b, Anaplasma