Kumar Ankeet, Tatu Utpal
Department of Biochemistry, Division of Biological Sciences, Indian Institute of Science, Bangalore, India.
Arch Microbiol. 2025 Jan 22;207(2):39. doi: 10.1007/s00203-025-04235-0.
Rabies is a deadly neurotropic, zoonotic disease with a mortality rate of 100% after symptoms appear. Rabies virus (RABV) is the primary cause of rabies disease in humans, and it mainly spreads via dog bites in developing countries. Over the course of RABV evolution, multiple RABV variants, called clades, have emerged. However, our understanding of these clades is limited, as the only method to identify a clade is sequencing, followed by phylogeny. In this study, we have developed a rapid, PCR-based method for typing two RABV clades. We utilised highly conserved amino acid changes specific to the Arctic and Africa-2 clades of the rabies virus (RABV). A single nucleotide substitution from adenine to thymine at position 178 within the nucleoprotein gene was found to be clade-specific in the Arctic clade. Similarly, adenine at position 638 is a distinctive marker for the Africa-2 clade. The assay demonstrated high specificity and offers the added benefit of PCR-based amplification, enabling virus detection even when viral titers are low. The assay can identify the Arctic clade and Africa-2 clade without sequencing and is highly specific and sensitive. Furthermore, this method can be adapted to detect other RABV clades and a wide range of viruses.
狂犬病是一种致命的嗜神经性人畜共患病,症状出现后死亡率为100%。狂犬病病毒(RABV)是人类狂犬病的主要病因,在发展中国家主要通过狗咬伤传播。在RABV的进化过程中,出现了多种RABV变体,称为进化枝。然而,我们对这些进化枝的了解有限,因为识别进化枝的唯一方法是测序,然后进行系统发育分析。在本研究中,我们开发了一种基于PCR的快速方法来分型两种RABV进化枝。我们利用了狂犬病病毒(RABV)北极进化枝和非洲2进化枝特有的高度保守氨基酸变化。在核蛋白基因内第178位的腺嘌呤到胸腺嘧啶的单核苷酸替换在北极进化枝中是进化枝特异性的。同样,第638位的腺嘌呤是非洲2进化枝的独特标记。该检测方法具有高特异性,并具有基于PCR扩增的额外优势,即使病毒滴度较低也能进行病毒检测。该检测方法无需测序即可识别北极进化枝和非洲2进化枝,且具有高度特异性和敏感性。此外,该方法可适用于检测其他RABV进化枝和多种病毒。