FAO Reference Center for Rabies, Istituto Zooprofilattico Sperimentale delle Venezie, 35020 Legnaro, PD, Italy.
Innovative Virology Laboratory, Research and Innovation Department, Istituto Zooprofilattico Sperimentale delle Venezie, 35020 Legnaro, PD, Italy.
Viruses. 2023 Mar 4;15(3):680. doi: 10.3390/v15030680.
Rabies is a zoonotic and fatal encephalitis caused by members of the genus. Among them, the most relevant species is , which is estimated to cause 60,000 human and most mammal rabies deaths annually worldwide. Nevertheless, all lyssaviruses can invariably cause rabies, and therefore their impact on animal and public health should not be neglected. For accurate and reliable surveillance, diagnosis should rely on broad-spectrum tests able to detect all known lyssaviruses, including the most divergent ones. In the present study, we evaluated four different pan-lyssavirus protocols widely used at an international level, including two real-time RT-PCR assays (namely LN34 and JW12/N165-146), a hemi-nested RT-PCR and a one-step RT-PCR. Additionally, an improved version of the LN34 assay ((n) LN34) was developed to increase primer-template complementarity with respect to all lyssavirus species. All protocols were evaluated in silico, and their performance was compared in vitro employing 18 lyssavirus RNAs (encompassing 15 species). The (n) LN34 assay showed enhanced sensitivity in detecting most lyssavirus species, with limits of detection ranging from 10 to 100 RNA copies/µL depending on the strain, while retaining high sensitivity against . The development of this protocol represents a step forward towards improved surveillance of the entire Lyssavirus genus.
狂犬病是一种由属成员引起的人畜共患和致命脑炎。其中，最相关的物种是 ，据估计，它每年在全球导致 60,000 例人类和大多数哺乳动物狂犬病死亡。然而，所有的狂犬病病毒都能引起狂犬病，因此不应忽视它们对动物和公共卫生的影响。为了进行准确和可靠的监测，诊断应依赖于能够检测所有已知狂犬病病毒的广谱测试，包括最具差异的病毒。在本研究中，我们评估了四种在国际上广泛使用的不同泛狂犬病病毒方案，包括两种实时 RT-PCR 检测方法（即 LN34 和 JW12/N165-146）、一种半巢式 RT-PCR 和一种一步法 RT-PCR。此外，还开发了 LN34 检测方法的改进版本（(n)LN34），以增加与所有狂犬病病毒物种的引物-模板互补性。所有方案均进行了计算机模拟评估，并通过体外试验比较了它们的性能，试验中使用了 18 种狂犬病病毒 RNA（涵盖 15 个种）。(n)LN34 检测方法显示出对大多数狂犬病病毒物种的检测灵敏度增强，检出限范围从 10 到 100 RNA 拷贝/µL 不等，具体取决于毒株，同时对 保留了高灵敏度。该方案的开发是朝着改进整个狂犬病属监测迈出的一步。
