Poxvirus and Rabies Branch, Division of High-Consequence Pathogens and Pathology, National Center for Emerging and Zoonotic Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, GA, United States.
J Virol Methods. 2013 Dec;194(1-2):229-34. doi: 10.1016/j.jviromet.2013.08.027. Epub 2013 Sep 12.
The genus Parapoxvirus (PAPV) is comprised traditionally of orf virus (ORFV), pseudocowpox virus (PCPV) and bovine papular stomatitis virus (BPSV), which cause infections of ruminants and their handlers in the U.S. and worldwide. Unlike orthopoxvirus infections, which can cause systemic or localized infections, PAPV infections present normally as benign, self-limited and localized skin lesions; infections do not confer lifelong immunity. In recent years, related potentially to enhanced awareness and the availability of diagnostic methods, there has been an observed increase in reported cases of PAPV in animals and humans. This study describes TaqMan based real-time PCR assays for both generic and specific detection of PAPV species for surveillance and outbreak investigations. These assays target highly conserved PAPV RNA polymerase gene sequences and are capable of detecting three known species of PAPVs (ORFV, PCPV, and BPSV). The assays were evaluated using a panel of PAPV DNA derived from human infections or animal specimen remainders. The sensitivities of all four assays were determined using droplet digital PCR; fewer than 10 copies of clinical PAPV DNA can be detected consistently. These assays provide a reliable and sensitive method for rapid confirmation and characterization PAPV infections with varying clinical presentations.
副痘病毒（PAPV）属传统上包含口疮病毒（ORFV）、假性牛痘病毒（PCPV）和牛丘疹性口炎病毒（BPSV），这些病毒在美国和全球范围内引起反刍动物及其饲养员的感染。与可引起全身或局部感染的正痘病毒感染不同，PAPV 感染通常表现为良性、自限性和局部皮肤损伤；感染不会产生终身免疫力。近年来，由于人们对 PAPV 的认识增强以及诊断方法的可用性提高，观察到动物和人类 PAPV 报告病例有所增加。本研究描述了用于 PAPV 种属通用和特异性检测的 TaqMan 实时 PCR 检测方法，用于监测和暴发调查。这些检测方法针对 PAPV RNA 聚合酶基因的高度保守序列，能够检测三种已知的 PAPV 种（ORFV、PCPV 和 BPSV）。使用源自人类感染或动物标本残余物的 PAPV DNA 进行了检测方法的评估。使用液滴数字 PCR 确定了所有四种检测方法的灵敏度；可以持续检测到少于 10 个拷贝的临床 PAPV DNA。这些检测方法为具有不同临床表现的 PAPV 感染的快速确认和特征分析提供了可靠和敏感的方法。
