用于敏感定量检测抗病毒 I 型干扰素的种属非依赖型生物测定法。

BACKGROUND

Studies of the host response to infection often require quantitative measurement of the antiviral type I interferons (IFN-alpha/beta) in biological samples. The amount of IFN is either determined via its ability to suppress a sensitive indicator virus, by an IFN-responding reporter cell line, or by ELISA. These assays however are either time-consuming and lack convenient readouts, or they are rather insensitive and restricted to IFN from a particular host species.

RESULTS

An IFN-sensitive, Renilla luciferase-expressing Rift Valley fever virus (RVFV-Ren) was generated using reverse genetics. Human, murine and avian cells were tested for their susceptibility to RVFV-Ren after treatment with species-specific IFNs. RVFV-Ren was able to infect cells of all three species, and IFN-mediated inhibition of viral reporter activity occurred in a dose-dependent manner. The sensitivity limit was found to be 1 U/ml IFN, and comparison with a standard curve allowed to determine the activity of an unknown sample.

CONCLUSIONS

RVFV-Ren replicates in cells of several species and is highly sensitive to pre-treatment with IFN. These properties allowed the development of a rapid, sensitive, and species-independent antiviral assay with a convenient luciferase-based readout.

背景

研究宿主对感染的反应通常需要定量测量生物样本中的抗病毒 I 型干扰素（IFN-α/β）。干扰素的数量可以通过其抑制敏感指示病毒的能力、通过干扰素反应报告细胞系或通过 ELISA 来确定。然而，这些测定方法要么耗时且缺乏方便的读数，要么灵敏度较低且仅限于来自特定宿主物种的 IFN。

结果

使用反向遗传学生成了一种 IFN 敏感的、表达 Renilla 荧光素酶的裂谷热病毒（RVFV-Ren）。在用种特异性 IFN 处理后，测试了人、鼠和禽细胞对 RVFV-Ren 的易感性。RVFV-Ren 能够感染这三种细胞，IFN 介导的病毒报告活性抑制呈剂量依赖性。灵敏度下限为 1 U/ml IFN，与标准曲线的比较允许确定未知样品的活性。