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Development of a Lightcycler-based reverse transcription polymerase chain reaction for the detection of foot-and-mouth disease virus
Journal of Veterinary Science ; : 207-212, 2005.
Artigo em Inglês | WPRIM | ID: wpr-128177
ABSTRACT
One step TaqMan real-time reverse transcription polymerase chain reaction (R/T RT-PCR)using a set of primers/probes was developed for the detection of foot-and-mouth disease (FMD)virus. The gene-specific probes labeled fluorogen for the internal ribosomal entry site, Leader sequence and 2B regions were used to detect FMD virus (FMDV). This assay specifically detected FMDV both in cell culture preparations and clinical samples, and was capable of distinguishing FMD from other viral diseases similar to clinical signs (swine vesicular disease, vesicular stomatitis and bovine viral diarrhea). This assay was shown to be 1000-fold more sensitive than the conventional RT-PCR method. The detection limits of this assay was 1 TCID 50 /ml of the FMDV RNA concentration. Quantification was obtained by a standard curves plotting threshold cycle values versus known infectivity titer. The assay was sensitive, specific and rapid enough to detect FMDV RNA genome in probang samples. As such, the described method is reliable and provides faster disease diagnostics than the conventional RT-PCR procedure to detect FMDV.
Assuntos

Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Sensibilidade e Especificidade / Taq Polimerase / Reação em Cadeia da Polimerase Via Transcriptase Reversa / Vírus da Febre Aftosa / Febre Aftosa Tipo de estudo: Estudo diagnóstico Limite: Animais Idioma: Inglês Revista: Journal of Veterinary Science Ano de publicação: 2005 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Sensibilidade e Especificidade / Taq Polimerase / Reação em Cadeia da Polimerase Via Transcriptase Reversa / Vírus da Febre Aftosa / Febre Aftosa Tipo de estudo: Estudo diagnóstico Limite: Animais Idioma: Inglês Revista: Journal of Veterinary Science Ano de publicação: 2005 Tipo de documento: Artigo