Duplex Real-Time Polymerase Chain Reaction for Simultaneous Detection and Quantification of Anaplasma Marginale and Anaplasma Centrale | Zendy

Nicola Decaro | Zendy; Grazia Carelli | Zendy; Eleonora Lorusso | Zendy; Maria Stella Lucente | Zendy; Grazia Greco | Zendy; Alessio Lorusso | Zendy; Arianna Radogna | Zendy; Luigi Ceci | Zendy; Canio Buonavoglia | Zendy

Open Access

Duplex Real-Time Polymerase Chain Reaction for Simultaneous Detection and Quantification of Anaplasma Marginale and Anaplasma Centrale

Author(s) -

Nicola Decaro,

Grazia Carelli,

Eleonora Lorusso,

Maria Stella Lucente,

Grazia Greco,

Alessio Lorusso,

Arianna Radogna,

Luigi Ceci,

Canio Buonavoglia

Publication year - 2008

Publication title -

journal of veterinary diagnostic investigation

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.529

H-Index - 78

eISSN - 1943-4936

pISSN - 1040-6387

DOI - 10.1177/104063870802000511

Subject(s) - anaplasma , anaplasmosis , polymerase chain reaction , biology , real time polymerase chain reaction , virology , microbiology and biotechnology , multiplex , hybridization probe , anaplasmataceae , nested polymerase chain reaction , dna , gene , genetics , tick

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.

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