RESUMO
Nested reverse transcription (RT)-PCR and SYBR Green real time PCR protocols for the intravitam detection of rabies virus genomic RNA were tested with clinical samples for the first time from rabies suspected animals (n=12). With SYBR Green real time PCR, five saliva samples were detected as positive, hence confirming that, for the sake of ante- mortem detection of rabies in saliva of animals, the sensitivity of real time PCR is more than that of RT-PCR as well as immunofluorescence that could detect rabies in three saliva samples each.(AU)
Assuntos
Animais , Raiva/diagnóstico , Saliva/virologia , Doenças Negligenciadas/veterinária , Reação em Cadeia da Polimerase em Tempo Real/métodosRESUMO
Molecular techniques were adopted to diagnose rabies viral RNA from skin biopsy samples collected from 20 animals. Nested RT-PCR and TaqMan real time PCR techniques have successfully diagnosed rabies viral RNA in 9 and 11 skin samples, respectively. The sensitivity of both techniques was calculated in comparison to FAT applied on brain samples. Sensitivity of 76.47% was obtained with nested RT-PCR on skin biopsy samples while Taqman real time PCR revealed sensitivity of 92.85%. It was concluded that TaqMan real time PCR is a useful, specific, sensitive and better molecular approach for antemortem rabies diagnosis from skin samples of rabies suspected animals.(AU)
Assuntos
Raiva/diagnóstico , Lyssavirus/isolamento & purificação , Epiderme/patologia , Biópsia/veterinária , Análise Citogenética/veterinária , Reação em Cadeia da Polimerase em Tempo Real/métodosRESUMO
Nested reverse transcription (RT)-PCR and SYBR Green real time PCR protocols for the intravitam detection of rabies virus genomic RNA were tested with clinical samples for the first time from rabies suspected animals (n=12). With SYBR Green real time PCR, five saliva samples were detected as positive, hence confirming that, for the sake of antemortem detection of rabies in saliva of animals, the sensitivity of real time PCR is more than that of RT-PCR as well as immunofluorescence that could detect rabies in three saliva samples each.
RESUMO
Molecular techniques were adopted to diagnose rabies viral RNA from skin biopsy samples collected from 20 animals. Nested RT-PCR and TaqMan real time PCR techniques have successfully diagnosed rabies viral RNA in 9 and 11 skin samples, respectively. The sensitivity of both techniques was calculated in comparison to FAT applied on brain samples. Sensitivity of 76.47% was obtained with nested RT-PCR on skin biopsy samples while Taqman real time PCR revealed sensitivity of 92.85%. It was concluded that TaqMan real time PCR is a useful, specific, sensitive and better molecular approach for antemortem rabies diagnosis from skin samples of rabies suspected animals.