RESUMEN
An indirect ELISA for the detection of japanese quail IgG specific to Newcastle disease virus (NDV) was developed. The secondary anti-quail IgG was produced in Balb/c mice, by inoculating Freund's complete adjuvant emulsified japanese quail-IgG extract. The purification of IgG was achieved using the caprilic acid method. The ELISA was compared to the haemagglutination-inhibition (HI) test for antibodies to NDV. ELISA cut-off point was established through TG-ROC analysis. Total correlation was observed between the ELISA and the HI, being the ELISA efficient in the identification of positive and negative sera, with high sensitivity and specificity (100 percent). These results validate the use of the indirect ELISA as an alternative for the detection of NDV-specific IgG in japanese quail sera, with the advantage of high sensitivity and automation
Asunto(s)
Animales , Coturnix , Ensayo de Inmunoadsorción Enzimática , Inmunoglobulina G/aislamiento & purificación , Virus de la Enfermedad de Newcastle/aislamiento & purificaciónAsunto(s)
Baculoviridae/genética , ADN Recombinante/análisis , Transfección , Animales , Células Cultivadas , ADN Recombinante/aislamiento & purificación , Desoxirribonucleasa EcoRI , Desoxirribonucleasas de Localización Especificada Tipo II , Vectores Genéticos , Reacción en Cadena de la Polimerasa , Spodoptera/metabolismoRESUMEN
A large number of DNA sequences corresponding to human and animal transcripts have been filed in data banks, as cDNAs or ESTs (expression sequence tags). However, the actual function of their corresponding gene products is still largely unknown. Several of these genes may play a role in regulation of important biological processes such as cell division, differentiation, malignant transformation and oncogenesis. Elucidation of gene function is based on 2 main approaches, namely, overexpression and expression interference, which respectively mimick or suppress a given phenotype. The currently available tools and experimental approaches to gene functional analysis and the most recent advances in mass cDNA screening by functional analysis are discussed.
Asunto(s)
Elementos sin Sentido (Genética) , Expresión Génica , ARN Catalítico , Transgenes , Animales , ADN Recombinante , HumanosRESUMEN
A large number of DNA sequences corresponding to human and animal transcripts have been filed in data banks, as cDNAs or ESTs (expression sequence tags). However, the actual function of their corresponding gene products is still largely unknown. Several of these genes may play a role in regulation of important biological processes such as cell division, differentiation, malignant transformation and oncogenesis. Elucidation of gene function is based on 2 main approaches, namely, overexpression and expression interference, which respectively mimick or suppress a given phenotype. The currently available tools and experimental approaches to gene functional analysis and the most recent advances in mass cDNA screening by functional analysis are discussed
