RESUMEN
Foot-and-mouth disease caused by foot-and-mouth disease virus (FMDV) is one of the most highly contagious diseases of domestic animals, and leads to enormous economic loss. Currently there are two main prevention and control strategies for the disease: eradication of the infected animals in FMDV free countries, and vaccination of the susceptible animals in countries with endemic FMDV infection. Early discovery and diagnosis of the source of infection is therefore integral to the containment of FMDV. In this study, a two-step reverse transcription recombinase polymerase amplification assay combined with lateral flow detection (RPA-LFD) was developed to detect FMDV. With incubation at 38 °C, a region of the 2B gene on the FMDV genome was successfully amplified within 20 min using specific primers and a probe. The amplified RPA product can be visualized on a lateral flow dipstick. The RPA-LFD assay was highly sensitive, detecting down to 10 copies of plasmid DNA. There was no cross-reactivity with other pathogens causing vesicular lesions. In addition, 143 clinical samples were used to compare RPA-LFD with real-time PCR, with 98.6% concordance between the assays. Therefore, the developed RPA-LFD assay provides a rapid, simple, highly promising approach to be used as point-of-care diagnostics in the field.
Asunto(s)
Virus de la Fiebre Aftosa/aislamiento & purificación , Fiebre Aftosa/diagnóstico , Inmunoensayo/métodos , Técnicas de Diagnóstico Molecular/métodos , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Animales , Animales Domésticos , Cartilla de ADN/genética , Virus de la Fiebre Aftosa/genética , Sondas de Oligonucleótidos/genética , Sensibilidad y Especificidad , Temperatura , Factores de TiempoRESUMEN
Bovine ephemeral fever virus (BEFV) is a typical species of the genusEphemerovirus in the family Rhabdoviridae. Today, prevailing BEFV can be divided into three phylogeographic lineages, East Asia, Mideast, and Australia. In this study, we provide evidence that the whole East Asia lineage originates from a homologous recombination (HR) between the Mideast and Australia lineages that probably occurred in the 1940s. To our knowledge, HR has not been proposed before as the genetic mechanism of BEFV. According to the HR event and Bayesian estimation, the three BEFV lineages might originate from Africa, and may have spread to Asia and Australia through the Mideast. In addition, the population of the virus may have augmented significantly in the 2000s, suggesting that the risk for outbreaks of BEFV may be high at present.
