massimo ancora - izs
TRANSCRIPT
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Massimo Ancora
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FINANZIAMENTO: Ministero della Salute, Dipartimento per la Sanità Pubblica Veterinaria, la Nutrizione e la Sicurezza degli Alimenti
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Gram –
Immobile
Aerobio
Asporigeno
Brucella sp.
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PCR real-time
Controllo interno di amplificazione (IAC)
Automazione
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PCR real time
7900 HT Fast PCR real-timeSystem (Applied Biosystems)
AmpliTaq Gold DNA Polymerase 3’-5’ TaqMan probe
Target diagnostico: IS711
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Maggiore sensibilità e specificità
Sequenza primer Brucella
Sequenza Brassica rapa esculenta
Sequenza probe Acetil CoA carbossilasi
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Maxwell® 16 Cell DNA purification kit (Promega)
MicroLab STARlet Robotic workstation – Hamilton
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6 da 2958 GE a 2,3 GE (4 replicati/diluizione)
: R2 tra 0,99 e 1 Efficienza >98% Coefficiente angolare tra -3,14 e -3,37
GE
Brucella abortus bv1
ufc/g
Brucella abortus bv3
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< Specificità
< Efficienza
< Sensibilità
< Sensibilità
No IAC
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192 campioni totali
19 campioni positivi batteriologico
41 campioni positivi PCR real-time
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Cammà C., Ancora M., Polci A., Di Provvido A., Di Giannatale E., Alessiani A., Lelli R. 2008. Development of a RealTime PCR method with an internal amplification control for high throughput detection of Brucella spp. Brucellosis 2008 International Research Conference. Royal Holloway College, University of London, UK, 10-13 September 2008, p.171 Di Provvido A., Ancora M., Cammà C., Catalani M., Scacchia M., Lelli R. 2008. Evaluation of RealTime PCR for a rapid diagnosis of Brucella spp. in cattle tissues. Brucellosis 2008 International Research Conference. Royal Halloway College, University of London, UK, 10-13 September 2008, p. 174
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Diagnosi precoce e specifica per la tutela della salute pubblica
Kit diagnostico commerciale
Future
Utilizzo presso il CVL, Windhoek (Namibia) NAPHL, Asmara (Eritrea)
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Grazie per l’attenzione