Quantitative Real-Time PCR Based on SYBR Green Technology for the Identification of Philaenus italosignus Drosopoulos & Remane (Hemiptera Aphrophoridae)

基于 SYBR Green 技术的定量实时 PCR 鉴别 Philaenus italosignus Drosopoulos & Remane(半翅目扁蚜科)

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作者:Domenico Rizzo, Matteo Bracalini, Sara Campigli, Anita Nencioni, Francesco Porcelli, Guido Marchi, Daniele Da Lio, Linda Bartolini, Elisabetta Rossi, Patrizia Sacchetti, Tiziana Panzavolta

Abstract

The use of molecular tools to identify insect pests is a critical issue, especially when rapid and reliable tests are required. We proposed a protocol based on qPCR with SYBR Green technology to identify Philaenus italosignus (Hemiptera, Aphrophoridae). The species is one of the three spittlebugs able to transmit Xylella fastidiosa subsp. pauca ST53 in Italy, together with Philaenus spumarius and Neophilaenus campestris. Although less common than the other two species, its identification is key to verifying which role it can play when locally abundant. The proposed assay shows analytical specificity being inclusive with different populations of the target species and exclusive with non-target taxa, either taxonomically related or not. Moreover, it shows analytical sensibility, repeatability, and reproducibility, resulting in an excellent candidate for an official diagnostic method. The molecular test can discriminate P. italosignus from all non-target species, including the congeneric P. spumarius.

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