Amphibians have been disappearing at an unprecedented rate worldwide. Among the proposed contributing factors are infectious diseases. Investigations have focused mainly on ranavirus and chytrids; however, additional agents may be relevant stressors. Two novel batrachoviruses have been discovered (ranid herpesvirus 3 [RaHV-3] and bufonid herpesvirus 1 [BfHV-1]). Their clinical role is still to be clarified; however, both have been associated with obvious skin lesions in their respective hosts. Herein we present 2 consensus PCR protocols that can be used to detect all of the known and, possibly, yet to be discovered batrachoviruses. We targeted a 200 nt long, highly conserved region of the DNA terminase gene. We established a sensitive protocol, which can detect both European batrachoviruses (European batrachovirus PCR protocol; RaHV-3 and BfHV-1) and a panbatrachovirus PCR protocol detecting all known batrachoviruses, including ranid herpesvirus 1 and 2 (RaHV-1, -2). The limit of detection (LOD) for the European batrachovirus protocol was 10¹ copies of RaHV-3 and 10² copies of BfHV-1 per reaction. The panbatrachovirus protocol could detect all known batrachoviruses with LODs of 10³ (RaHV-3, BfHV-1, RaHV-1) to 10⁴ copies (RaHV-2) per reaction. These novel detection tools can be used as a first line of detection when herpesviral infection in amphibians is suspected, followed by additional PCRs with herpesvirus-specific primers in the case of known viral species, or sequencing as in the case of novel batrachoviruses.

两栖动物在世界范围内以前所未有的速度消失。提议的促成因素包括传染病。调查主要集中在雷纳病毒和壶菌上；然而，额外的因素可能是相关的压力源。已经发现了两种新型蝙蝠病毒（ranid herpesvirus 3 [RaHV-3] 和 bufonid herpesvirus 1 [BfHV-1]）。它们的临床作用仍有待阐明；然而，两者都与各自宿主的明显皮肤损伤有关。在此，我们提出了 2 种共有 PCR 协​​议，可用于检测所有已知的和可能尚未发现的 batrachoviruses。我们的目标是 DNA 终止酶基因的一个 200 nt 长、高度保守的区域。我们建立了一个敏感的协议，可以检测两种欧洲蝙蝠病毒（European batrachovirus PCR protocol; RaHV-3 和 BfHV-1) 和检测所有已知 batrachoviruses 的 panbatrachovirus PCR 协​​议，包括ranid herpesvirus 1 和 2 (RaHV-1, -2)。欧洲 batrachovirus 协议的检测限 (LOD) 为 10^1个RaHV-3的副本和10个²拷贝BfHV-1每个反应。panbatrachovirus 协议可以检测所有已知的 batrachovirus，每个反应的LOD 为 10 ³ (RaHV-3、BfHV-1、RaHV-1) 到 10 ⁴拷贝 (RaHV-2)。当怀疑两栖动物的疱疹病毒感染时，这些新型检测工具可用作第一线检测，然后在已知病毒种类的情况下使用疱疹病毒特异性引物进行额外的 PCR，或在新型 batrachovirus 的情况下进行测序。