Human rabies post mortem diagnostic samples are often preserved in formalin. While immunohistochemistry (IHC) has been routinely used for rabies antigen detection in formalin-fixed tissue, the formalin fixation process causes nucleic acid fragmentation that may affect PCR amplification. This study reports the diagnosis of rabies in an individual from the Dominican Republic using both IHC and the LN34 pan-lyssavirus real-time RT-PCR assay on formalin-fixed brain tissue. The LN34 assay generates a 165 bp amplicon and demonstrated higher sensitivity than traditional PCR. Multiple efforts to amplify nucleic acid fragments larger than 300 bp using conventional PCR were unsuccessful, probably due to RNA fragmentation. Sequences generated from the LN34 amplicon linked the case to the rabies virus (RABV) strain circulating in the Ouest Department of Haiti to the border region between Haiti and the Dominican Republic. Direct sequencing of the LN34 amplicon allowed rapid and low-cost rabies genetic typing.

中文翻译：

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使用LN34泛狂犬病病毒实时RT-PCR检测狂犬病的诊断和福尔马林固定的人脑组织的快速基因分型。

验尸后的人类狂犬病诊断样品通常保存在福尔马林中。尽管免疫组织化学（IHC）已常规用于福尔马林固定组织中的狂犬病抗原检测，但福尔马林固定过程会导致核酸断裂，从而影响PCR扩增。这项研究报告了在福尔马林固定的脑组织上同时使用IHC和LN34泛狂犬病病毒实时RT-PCR测定法对多米尼加共和国个人的狂犬病诊断。LN34分析产生了165 bp的扩增子，并显示出比传统PCR更高的灵敏度。使用常规PCR扩增多于300 bp的核酸片段的多次尝试均未成功，这可能是由于RNA片段化所致。从LN34扩增子产生的序列将病例与狂犬病病毒（RABV）菌株联系起来，该病毒在海地西部部海地和多米尼加共和国之间的边界地区流通。LN34扩增子的直接测序可实现快速低成本的狂犬病基因分型。