Abstract：
Pentastiridius leporinus (Hemiptera: Cixiidae) is the main vector of an emerging and fast spreading sugar beet disease, the syndrome \'basses richesses\' (SBR), in different European countries. The disease is caused by the γ-3-proteobacterium \'Candidatus Arsenophonus phytopathogenicus\' and the phytoplasma \'Candidatus Phytoplasma solani\' which are exclusively transmitted by planthoppers and can lead to a significant loss of sugar content and yield. Monitoring of this insect vector is important for disease management. However, the morphological identification is time consuming and challenging as two additional cixiid species Reptalus quinquecostatus and Hyalesthes obsoletus with a very close morphology have been reported in sugar beet fields. Further, identification of females and nymphs of P. leporinus at species level based on taxonomic key is not possible. In this study, an isothermal nucleic acid amplification based on recombinase polymerase amplification (RPA) was developed to specifically detect P. leporinus. In addition, real-time RPA was developed to detect both adults (male and female) and nymph stages using pure or crude nucleic acid extracts. The sensitivity of the real-time RPA for detection of P. leporinus was comparable to real-time PCR, but a shorter time (< 7 min) was required. This is a first report for real-time RPA application for P. leporinus detection using crude nucleic acid templates which can be applied for fast and specific detection of this vector in the field.
摘要：
Pentastiridiusleporinus（半翅目：Cixiidae）是一种新兴且快速传播的甜菜病的主要媒介，“贝斯富有”综合征(SBR)，在不同的欧洲国家。该病是由γ-3-变形杆菌“念珠菌”和“念珠菌”引起的，它们仅由飞虱传播，可导致糖含量和产量的显着损失。监测这种昆虫媒介对于疾病管理很重要。然而,形态学鉴定是耗时且具有挑战性的，因为在甜菜田中已经报道了另外两个形态非常接近的cixiid物种Reptalusquinquecostatus和Hyalesthes过时。Further,根据分类学关键在物种水平上鉴定假单胞菌的雌性和若虫是不可能的。在这项研究中,开发了一种基于重组酶聚合酶扩增（RPA）的等温核酸扩增技术来特异性检测假单胞菌。此外,使用纯或粗核酸提取物开发了实时RPA来检测成年（男性和女性）和若虫阶段。实时RPA的灵敏度与实时PCR相当。但需要更短的时间（<7分钟）。这是使用粗核酸模板实时RPA应用于小孢子菌检测的第一份报告，该模板可用于该载体的快速和特异性检测。
