Development and Field Testing of a Real-Time PCR Assay for Caprine Arthritis-Encephalitis-Virus (CAEV)

Brajon G;Mandas D;Liciardi M;Taccori F;MELONI, MAURO;Corrias F;Montaldo C;Coghe F;Casciari C;Giammarioli M;ORRU, GERMANO

2012-01-01

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Abstract

Abstract: caprine arthritis/encephalitis (CAE) of goats and occasionally sheep, are persistent virus infections caused by a lentivirus (CAEV). This viral infection results in arthritis in adult animals and encephalitis in kids. Prognosis for the encephalitic form is normally poor, with substantial economic loss for the farm. In this context an early/fast laboratory diagnosis for CAEV infection could be useful for effective prophylactic action. In this work we performed a quantitative real time PCR designed on the CAEV env gene to detect/quantify in goat/sheep samples, viral RNA or proviral DNA forms of CAEV. This procedure was validated in 15 sheep experimentally infected with CAEV or with a high correlated lentivirus ( visna maedi, MVV) ; in addition, a total of 37 clinical goat specimens recruited in CAEV positive herds were analyzed and compared using serological analysis (Elisa and AGID). All samples infected with MVV resulted negative. In sheep, experimentally infected with CAEV , proviral DNA was detectable 15 days post infection, whereas the serological methods revealed an indicative positivity after 40-60 days. This method showed a sensitivity of 102 env fragments /PCR) with a linear dynamic range of quantitation from 102 to 107 env fragments/PCR, the correlation coefficient R2 was 0.98. All subjects with clinical diagnosis for Caprine Arthritis-Encephalitis (CAE) resulted CAEV DNA positive.