A viral pseudotype based approach to measuring morbillivirus neutralising antibodies

该项目的目标是开发一种快速、敏感和特异的诊断测试。这项测试将被用于诊断一种名为PPR的绵羊和山羊的病毒感染，或小反刍动物的害虫。PPR是对全世界畜牧业的威胁，已被宣布为通过接种疫苗进行全球控制和根除的优先目标。要想成功地控制和接种小反刍动物病毒(PPRV)的全球控制和疫苗接种方案，将需要提供一种高度准确和快速的诊断测试，这种测试无需昂贵的专门实验室隔离设施，因为必须在世界各地的实验室进行。然而，目前对猪繁殖与呼吸综合征病毒的规定测试是活病毒中和测试，这是一种缓慢的测试，需要实验室在高水平的生物遏制下操作，并且在敏感性和灵活性方面都受到限制。为了发展一种改进的PPRV诊断试验，我们使用了一种技术，即在水疱性口炎病毒(VSV)来源的无害载体病毒上表达PPRV表面糖蛋白，以产生VSV(PPRV)“假型”。病毒伪型由一种病毒(在这种情况下是VSV)的结构蛋白和遗传物质组成，携带另一种不同病毒的表面蛋白(在本实验中我们使用PPRV H和F蛋白)。VSV(PPRV)伪型可用于检测PPRV特异性抗体，比传统的活病毒检测更快(3天，而不是1-2周)。该检测方法显示出比活病毒测试更高的灵敏度(至少是活病毒测试的10倍)，并可以测量对代表不同病毒谱系的不同病毒株的抗体反应。最后，该分析避开了对高水平生物遏制的要求，可以在世界各地任何拥有基本细胞培养设施的实验室安全地进行。在这个项目中，我们将通过生成基于VSV(PPRV)的诊断测试来协助世界动物卫生组织的全球PPRV控制和根除计划。通过这项测试，我们将能够识别受感染的动物，并调查可能导致该病毒传播给牲畜的野生动物种群。这些信息将有助于确保疫苗接种方案针对传播风险高的地区的易感人群，从而提高成功根除猪繁殖与呼吸综合征病毒的机会。

The goal of this project is to develop a diagnostic test that is rapid, sensitive and specific. This test will be used to diagnose a viral infection of sheep and goats called PPR, or peste des petits ruminants. PPR is a threat to the livestock industry worldwide and has been declared a priority target for global control and eradication by vaccination. The success of a global control and vaccination programme against peste des petits ruminants virus (PPRV) will require the availability of a highly accurate and rapid diagnostic test that can be performed without the need for expensive specialized laboratory containment facilities since it will have to be carried out in laboratories across the world. However, the current prescribed test for PPRV is the live virus neutralisation test, a test that is slow, requires a laboratory operating under a high level of biocontainment, and is limited in both sensitivity and flexibility. In order to develop an improved PPRV diagnostic test, we have used a technique in which PPRV surface glycoproteins are expressed on a harmless carrier virus derived from vesicular stomatitis virus (VSV) to generate VSV(PPRV) "pseudotypes". A viral pseudotype consists of the structural proteins and genetic material of one virus (in this case VSV), carrying the surface proteins of another, different virus (in this assay we use the PPRV H and F proteins). The VSV(PPRV) pseudotypes can be used to measure PPRV specific antibodies more rapidly than conventional live virus tests (three days compared with 1 to 2 weeks). The assay displays greater sensitivity (at least 10-times more sensitive) than live virus tests and can measure antibody responses against diverse strains of virus representative of distinct viral lineages in circulation. Finally, the assay circumvents the requirement for high level biocontainment and may be performed safely in any laboratory worldwide with basic cell culture facilities.In this project we will assist the World Animal Health Organization's Global Control and Eradication program for PPRV by generating a VSV(PPRV)-based diagnostic test. With this test, we will be able to identify infected animals and investigate populations of wild animals which may be responsible for transmitting this virus to livestock. This information will help ensure that vaccination programmes target susceptible populations in areas with a high risk of transmission and therefore improve the chances of successfully eradicating PPRV.