Kyasanur Forest disease virus (KFDV) and Alkhurma hemorrhagic fever virus (AHFV) are enveloped, positive-stranded RNA viruses of clinical importance with complex enzootic life cycles involving hematophagous ticks which feed on small and large mammals. Humans and monkeys are dead-end hosts for these viruses. Recent trends in epidemiological data suggest both virus incidences are steadily increasing and their geographical distribution expanding out of previously known circulation regions. For the detection and discrimination of these two closely related flaviviruses, we have developed quantitative real-time RT-PCR assays with 100 % sensitivity and 100 % specificity for KFDV, 100 % sensitivity and 99.4 % specificity for AHFV as determined using 550 clinical samples collected between 2015–2018 from Western Ghats region of India. This rapid and sensitive assay will enable researchers to accurately diagnose the presence of the virus during viremia in human and animal blood samples, and also from tick specimens. Incorporation of these new tests into a routine diagnosis will help in the diagnosis of KFDV as well as AHFV in the endemic areas and also would provide an early warning of the spread of this virus to newer regions with similar epidemiology.

Kyasanur森林病病毒（KFDV）和Alkhurma出血热病毒（AHFV）是具有临床重要性的有包膜正链RNA病毒，具有复杂的生化生命周期，涉及以大小食者为食的he虫。人类和猴子是这些病毒的死胡同。流行病学数据的最新趋势表明，这两种病毒的发病率都在稳步增加，其地理分布也从先前已知的流行区域扩展。为了检测和区分这两种密切相关的黄病毒，我们开发了定量实时RT-PCR分析，对KFDV的敏感性为100％，特异性为100％，对AHFV的敏感性为100％，特异性为99.4％（使用收集的550份临床样品测定） 2015年至2018年之间，来自印度西高止山脉地区。这种快速而灵敏的测定方法将使研究人员能够准确地诊断人和动物血样以及壁虱标本在病毒血症期间的病毒存在。将这些新检测方法纳入常规诊断将有助于在流行地区诊断KFDV以及AHFV，也将提供该病毒向具有类似流行病学特征的较新区域传播的预警。