Duplex real-time polymerase chain reaction for simultaneous detection and quantification of Anaplasma marginale and Anaplasma centrale.
Anaplasma marginale and Anaplasma centrale are rickettsial pathogens responsible for acute disease and mild infections, respectively, in cattle herds. A duplex real-time polymerase chain reaction (PCR) assay with probes labeled with different fluorophores was developed for simultaneous detection and quantification of A. marginale and A. centrale DNA in bovine blood samples. The assay was able to detect as few as 10(1) and 10(2) DNA copies for A. marginale and A. centrale, respectively, with optimal specificity and reproducibility. Analysis by real-time and nested PCR carried out on 54 samples previously tested by reverse line blot hybridization showed that the established duplex real-time PCR assay can detect and quantify the 2 Anaplasma spp., even if present simultaneously in the same blood samples. Such an assay could be used in pathogenesis studies on bovine acute anaplasmosis.
Decaro
Nicola
N
Department of Public Health and Zootechnics, Faculty of Veterinary Medicine, University of Bari, Valenzano, Bari, Italy. n.decaro@veterinaria.uniba.it
Carelli
Grazia
G
Lorusso
Eleonora
E
Lucente
Maria Stella
MS
Greco
Grazia
G
Lorusso
Alessio
A
Radogna
Arianna
A
Ceci
Luigi
L
Buonavoglia
Canio
C
D042321:Anaplasma centrale / Q000235:genetics* / Q000302:isolation & purification; D042322:Anaplasma marginale / Q000235:genetics* / Q000302:isolation & purification; D000712:Anaplasmosis / Q000175:diagnosis*; D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000382:microbiology*; D003937:Diagnosis, Differential; D015511:Goat Diseases / Q000175:diagnosis / Q000382:microbiology; D006041:Goats; D007558:Italy; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012756:Sheep; D012757:Sheep Diseases / Q000175:diagnosis / Q000382:microbiology