RESUMEN
A chromatographic strip assay was developed for rapid detection of serum antibodies to non-structural protein of foot-and-mouth disease virus. The assay was based on Escherichia coli-expressed 3ABC non-structural protein and an immunochromatographic technique, which shortened the detection time to about one hour. The sensitivity of the assay was determined to be 96.8% for infected pigs; its specificity was 100% for naïve pigs and 98.8% for vaccinated pigs. In the experimentally infected pigs, anti-3ABC antibodies were detectable from eight days post-infection until the end of the study, 34 days post-infection. The performance of this assay was comparable to that of two commercial ELISA kits, Ceditest FMDV-NS and UBI FMDV NS EIA, and was better than that of CHEKIT FMD-3ABC po. Given its advantages of instant testing and quantitative measurement, this assay has potential as a useful tool for rapid on-farm diagnosis of foot-and-mouth disease.
Asunto(s)
Anticuerpos Antivirales/sangre , Virus de la Fiebre Aftosa/inmunología , Fiebre Aftosa/diagnóstico , Enfermedades de los Porcinos/inmunología , Proteínas no Estructurales Virales/inmunología , Animales , Clonación Molecular , Escherichia coli/genética , Escherichia coli/metabolismo , Fiebre Aftosa/virología , Virus de la Fiebre Aftosa/genética , Virus de la Fiebre Aftosa/aislamiento & purificación , ARN Viral/química , ARN Viral/genética , Tiras Reactivas , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Sensibilidad y Especificidad , Porcinos , Enfermedades de los Porcinos/diagnóstico , Enfermedades de los Porcinos/virología , Proteínas no Estructurales Virales/biosíntesis , Proteínas no Estructurales Virales/genéticaRESUMEN
For investigating the feasibility of using reactive dyes as an immunoassay marker, a dichlorine triazine dye, Procion Blue MX-7RX, was employed to stain the antibody against human serum albumin (anti-HSA). With the color intensity revealed in the immunochromatographic test strip as the objective variables, the optimal dyeing conditions were found as follows: pH 11.4, temperature 35.7 degrees C, molar ratio 188 (mol dye/mol antibody), and reaction time 45.6 min. The dyed-anti-HSA revealed a maximal color intensity of 8738 without apparent loss of antigen binding affinity.
