Department of Plant Protection, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran.
Biodiversity & Conservation, Manaaki Whenua Landcare Research, Lincoln, New Zealand.
PLoS One. 2018 Oct 4;13(10):e0205136. doi: 10.1371/journal.pone.0205136. eCollection 2018.
Tephritid fruit flies are ranked as one of the most damaging groups of insect pests. Morphological identification of fruit flies is mainly performed on adults due to the lack of adequate identification keys for immature stages. The peach fruit fly, Bactrocera zonata (Saunders), infests some of the principal commercial fruits and vegetables. It is, therefore important to avert its global dispersal, particularly by accurately identifying this species at ports of entry. In this study, a TaqMan real-time polymerase chain reaction (PCR) was developed for the accurate identification and sensitive detection of the peach fruit fly. A novel set of primers and probe were designed to specifically identify the mitochondrial cytochrome oxidase I (COI) gene. All specimens of peach fruit fly (including various life stages) were detected, and no cross reactivity with other tested tephritids were observed. Since this assay performed equally well with crushed insects and purified DNA, we note added efficiency by eliminating DNA extraction step. Considering the speed, specificity as well as sensitivity of the assay, Taqman real-time PCR can be used as a swift and specific method for pest species at ports of entry.
实蝇科水果蝇被列为最具破坏性的昆虫害虫之一。由于缺乏成熟阶段的充分鉴定标准,水果蝇的形态鉴定主要在成虫期进行。桃实蝇,Bactrocera zonata(桑德斯),侵害一些主要的商业水果和蔬菜。因此,防止其在全球范围内扩散非常重要,特别是要在入境口岸准确识别这种物种。在这项研究中,开发了一种 TaqMan 实时聚合酶链反应(PCR),用于准确识别和灵敏检测桃实蝇。设计了一套新的引物和探针,用于特异性识别线粒体细胞色素氧化酶 I(COI)基因。所有桃实蝇标本(包括各种生命阶段)均被检测到,与其他测试的实蝇科昆虫没有交叉反应。由于该检测方法对粉碎昆虫和纯化 DNA 的效果相同,因此我们注意到通过消除 DNA 提取步骤提高了效率。鉴于该检测方法的速度、特异性和灵敏度,Taqman 实时 PCR 可用于入境口岸的害虫物种。