![]() Therefore, it is often necessary to rely on laboratory diagnosis. However, the clinical symptoms of infection with ORFV can be remarkably similar to those of other viruses (such as sheep pox virus and goat pox virus), making differential diagnosis difficult. ORFV can be routinely diagnosed on the basis of typical skin lesions, clinical signs, and epidemic symptoms. Orf is a zoonotic disease, and farm workers and others who have direct or indirect contact with ORFV-infected animals can occasionally become infected with ORFV through skin wounds. Orf outbreaks have occurred in many countries, causing huge economic losses to the breeding industry. Infected sheep experience decreased feed intake, slow growth and development, and rapid spread throughout the breeding area, causing serious economic losses to the farm. Symptoms such as muzzle skin or mucous membrane ulcers, papules, abscesses, scabs, and other symptoms are characteristic of ORFV infection in sheep. Lambs aged 1–6 months are most susceptible to ORFV infection. ORFV can cause infectious pustulosis in goats and sheep and is widespread in most sheep-raising countries in the world. It belongs to the genus parapoxvirus of the family Poxviridae. ORFV is an epitheliophilic double-stranded DNA virus. Compared with orthopoxvirus, paraxxvirus is relatively small in size and has a high content of G C in the genome. The members of parapoxvirus include Orf virus (ORFV), bovine papular stomatitis virus (BPSV), pseudocowpox virus (PCPV), and the newly discovered parapoxvirus of red deer in New Zealand (PVNZ). Parapoxvirus belongs to Chordopoxvirinae, a vertebrate Poxviridae, and is one of the most commonly studied genera of poxvirus. Poxvirus genomes evolved through the acquisition and loss of genes, especially gene replication and horizontal gene transfer, to adapt to and combat the host’s antiviral immune defenses to establish successful infection. The origin and evolution of poxviruses remain obscure, despite evidence that poxviruses emerged thousands of years ago. Recently, increasing attention has been given to it. Poxviridae is a widely known virus family with the largest number of members of any virus it is the largest and most complex virus known to harm mammals. The DNA extraction–free visual detection method of RAA established in this study can meet the needs of rapid onsite detection and grassroots laboratories and has important reference value and significance for the early diagnosis of diseased animals. Compared with qPCR, the sensitivity and specificity of the real-time RAA assay were 100%, and those of the visual RAA assay were 92.31% and 100.0%, respectively. The detection limits of the real-time RAA and visual judgment of the RAA assay at 95% probability were 13 and 21 copies per reaction for ORFV, respectively. This method is specific to ORFV and does not cross-react with other common DNA viruses. This study establishes a DNA extraction–free, real-time, visual recombinase–aided amplification (RAA) method for the rapid detection of ORFV. Although diagnostic methods for ORFV infection already exist, it is still necessary to develop a time-saving, labor-saving, specific, low-cost and visual diagnostic method for rapid detection of ORFV in the field and application in grassroots laboratories. Orf is widespread in most sheep-raising countries in the world, causing huge economic losses. kagit geri donusum ornekleri stream hÞ¬[[Sãƶþ zLj×6}¿Tíš* Æ`l ƒSy ŒgŽO ˜â²'9¿þ|«WËØ
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