RESUMEN
The Reticuloendotheliosis virus (REV) group of retroviruses infects a wide range of avian species, including chickens, turkeys, ducks, geese, quail, and prairie chickens. The objective of the present study was to develop a highly sensitive and specific diagnostic test for the detection of REV in whole blood samples. In order to increase the diagnostic sensitivity, a duplex real-time polymerase chain reaction (PCR) that detects both the envelope protein gene (env) and the long terminal repeat (LTR) region of REV was designed. This assay demonstrated greater analytical and diagnostic sensitivity than the gel-based PCR assay when using DNA extracted from whole blood by both phenol-chloroform and magnetic bead methods. In general, threshold cycle values in the duplex real-time PCR assay were lower from DNA extracted using the magnetic bead system compared to DNA extracted by the phenol-chloroform method. Data presented herein show the successful development of a rapid and accurate test procedure, with high-throughput capability, for the diagnosis of REV infection using avian blood samples.
Asunto(s)
Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Virus de la Reticuloendoteliosis Aviar/aislamiento & purificación , Reticuloendoteliosis Aviar/diagnóstico , Secuencias Repetidas Terminales/genética , Proteínas del Envoltorio Viral/genética , Animales , Fraccionamiento Químico/métodos , ADN Viral/genética , Patos , Galliformes , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Reticuloendoteliosis Aviar/sangre , Reticuloendoteliosis Aviar/virología , Sensibilidad y EspecificidadRESUMEN
Two commercial flocks of Chinese partridge experienced increased mortality associated with a wasting disease at 120-day old in June 2006. Postmortem examination of dead chickens consistently showed visceral tissues mainly proventriculus, liver and spleen were diffuse enlargement. Microscopic examination revealed masses of immature lymphocytes with frequent mitotic figures were seen in various tissues including proventriculus, liver, spleen, kidney, heart, lung, thymus and intestine. Reticuloendotheliosis virus (REV) was isolated from each of four blood samples. Viral antigens were observed in cultured CEF (SPF embryos came from the Ji-nan poultry institute) inoculated blood samples via on indirect immunofluorescent assay. Three hundred bp fragments of LTR of REV obtained from liver samples of six chickens by PCR. This disease has not previously been reported in Chinese partridge. Chinese partridge may represent a potential reservoir of infection for other Chinese local chickens.
Asunto(s)
Galliformes , Enfermedades de las Aves de Corral/epidemiología , Enfermedades de las Aves de Corral/patología , Reticuloendoteliosis Aviar/veterinaria , Animales , China/epidemiología , Femenino , Riñón/citología , Riñón/patología , Hígado/patología , Linfocitos , Ovario/patología , Enfermedades de las Aves de Corral/diagnóstico , Enfermedades de las Aves de Corral/virología , Virus de la Reticuloendoteliosis Aviar/aislamiento & purificación , Reticuloendoteliosis Aviar/sangre , Reticuloendoteliosis Aviar/epidemiología , Reticuloendoteliosis Aviar/virología , Bazo/patologíaRESUMEN
Reticuloendotheliosis virus (REV) is widespread in the world. No related data has been reported in Taiwan. To determine the REV infection status, antibody determination and virus isolation were performed on chickens in Taiwan. The results revealed that serological flock prevalence for the REV antibody reached 92.8% (39/42) amongst breeders (> 16 weeks old). Two different REV isolates were identified by reverse transcriptase-polymerase chain reaction, electron microscopic, immunofluorescent, and western blot assays after isolation. One of these viruses was isolated from a broiler breeder farm and the other was isolated from a Taiwan Country Chicken farm. Despite their different origins, the percent identity of the nucleotide sequences of the env gene of these two isolates was 99.7%. These two strains were similar to the FPV-UI-REV strain, featuring 99.7% and 99.8% percent identity. Indeed, REV infection would appear to be quite common amongst chickens.