School of Clinical Veterinary Sciences, University of Bristol, Langford House, Langford, North Somerset, UK.
Exp Parasitol. 2010 Dec;126(4):506-9. doi: 10.1016/j.exppara.2010.05.026. Epub 2010 May 31.
Polymerase chain reaction (PCR) analysis is regularly used to detect pathogens within arthropod vectors, but has also been applied to investigate vector DNA. This study details a novel highly sensitive quantitative PCR (qPCR) which detects and quantifies DNA from Ixodes ricinus, the European vector of Anaplasma phagocytophilum. By pairing this with a qPCR to detect A. phagocytophilum, valid comparisons of pathogen load can be made between different sized tick-tissue samples. These qPCRs were validated in I. ricinus that were fed A. phagocytophilum-infected blood using an artificial membrane feeder. Pathogens were detected in the tick haemolymph within 36h, indicating that successful infection had taken place. This study illustrates the application of vector-targeted qPCRs to confirm and validate pathogen load in samples as part of investigations of vector-pathogen interactions.
聚合酶链式反应（PCR）分析通常用于检测节肢动物载体中的病原体，但也已应用于研究载体 DNA。本研究详细介绍了一种新型的高灵敏度定量 PCR（qPCR），可检测和定量欧洲蜱传阿尼姆斯噬细胞体的载体——硬蜱的 DNA。通过将其与检测阿尼姆斯噬细胞体的 qPCR 配对，可以在不同大小的蜱组织样本之间对病原体载量进行有效比较。这些 qPCR 在使用人工膜饲养器喂食感染阿尼姆斯噬细胞体的血液的硬蜱中得到了验证。在 36 小时内，在蜱的血淋巴中检测到病原体，表明已经成功感染。本研究说明了针对载体的 qPCR 在确认和验证样本中病原体载量方面的应用，这是对载体-病原体相互作用进行调查的一部分。
