基于探针的实时定量 PCR 和 RPA-Cas12a 分子诊断检测番茄鳞翅目害虫烟粉虱（Phthorimaea absoluta）。

Probe-based quantitative PCR and RPA-Cas12a molecular diagnostics for detection of the tomato pest Phthorimaea absoluta (Lepidoptera: Gelechiidae).

机构信息

Department of Entomology and Nematology, College of Agricultural and Environmental Sciences, University of California Davis, One Shields Ave, Davis, CA 95616, USA.

Contained Research Facility, University of California, Davis, 555 Hopkins Rd, Davis, CA 95616, USA.

出版信息

J Econ Entomol. 2023 Jun 13;116(3):993-1001. doi: 10.1093/jee/toad079.

DOI:10.1093/jee/toad079

PMID:37177893

Abstract

The tomato pest Phthorimaea absoluta Meyrick is highly invasive but has not yet invaded North America. However, several morphologically similar species are already present, making detection of P. absoluta presence and invasion challenging. We designed a quantitative PCR molecular diagnostic to differentiate P. absoluta, P. operculella (Zeller), or Keiferia lycopersicella (Walsingham) (Lepidoptera: Gelechiidae) DNA. Additionally, we developed an RPA-Cas12a molecular diagnostic that allows for the isothermal detection of P. absoluta DNA, eliminating the need for a thermocycler. The results of the RPA-Cas12a diagnostic can be visualized simply using a UV light source and cell phone camera. We expect these diagnostics to improve quarantine and prevention measures against this serious agricultural threat.

摘要

番茄潜叶蛾 Phthorimaea absoluta Meyrick 具有很强的入侵性，但尚未入侵北美。然而，已有几种形态相似的物种存在，这使得检测番茄潜叶蛾的存在和入侵变得具有挑战性。我们设计了一种定量 PCR 分子诊断方法，用于区分番茄潜叶蛾、卷叶蛾（Zeller）或番茄夜蛾（Keiferia lycopersicella）（鳞翅目：草螟科）的 DNA。此外，我们开发了一种 RPA-Cas12a 分子诊断方法，允许等温检测番茄潜叶蛾的 DNA，无需使用热循环仪。RPA-Cas12a 诊断的结果可以简单地使用紫外光源和手机摄像头进行可视化。我们期望这些诊断方法能够改进针对这一严重农业威胁的检疫和预防措施。