Orf, caused by orf virus (ORFV), is an important zoonotic disease that infects goat and sheep, leading to huge economic losses. ORFV can also cause cutaneous lesions in people who come in close contact with the diseased animals. Although accurate diagnostic methods for ORFV infection exist, there is a need for a rapid, specific, and sensitive method for easy clinical application. Here, we successfully established a recombinase-aided amplification (RAA) assay for rapid detection of ORFV. The analytical sensitivity of the assay for ORFV detection is 1 × 101 copies per reaction. Moreover, no cross-reaction was observed with other common DNA viruses. A total of 45 archived suspected ORFV infected nasal scab skin samples were examined by RAA and SYBR Green-based real-time polymerase chain reaction (PCR). Compared with the real-time PCR assay, the kappa values of the RAA assay for ORFV detection was 0.845 (p <0.001), indicating that both assay results were fully in agreement. In conclusion, this detection assay provides a rapid, sensitive, and specific method for ORFV detection and is suitable for ORFV clinical testing.

中文翻译：

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开发用于检测orf病毒的重组酶辅助扩增测定法。

由orf病毒（ORFV）引起的Orf是一种重要的人畜共患病，感染山羊和绵羊，导致巨大的经济损失。ORFV还可导致与患病动物密切接触的人的皮肤损害。尽管存在用于ORFV感染的准确诊断方法，但是需要快速，特异性和灵敏的方法以易于临床应用。在这里，我们成功地建立了用于快速检测ORFV的重组酶辅助扩增（RAA）分析。每个反应的ORFV检测分析灵敏度为1×101拷贝。此外，未观察到与其他常见DNA病毒的交叉反应。通过RAA和基于SYBR Green的实时聚合酶链反应（PCR）检查了总共45份怀疑是ORFV感染的鼻sc皮肤感染样本。与实时PCR分析相比，用于ORFV检测的RAA分析的kappa值为0.845（p <0.001），表明两个分析结果完全吻合。总而言之，这种检测方法为ORFV检测提供了一种快速，灵敏且特异的方法，适用于ORFV临床测试。