Phytopathology. 1999 Jun;89(6):522-8. doi: 10.1094/PHYTO.1999.89.6.522.
ABSTRACT Immuno-capillary zone electrophoresis (I-CZE) is a technique that combines the specificity afforded by serological assays with the sensitivity, rapidity, and automation in detection provided by capillary zone electrophoresis. Cymbidium mosaic potexvirus (CymMV) and odontoglossum ringspot tobamovirus (ORSV) were detected in their purified forms as well as in the crude saps of infected Nicotiana benthamiana leaves and Oncidium orchid flowers. The two orchid virus-antibody complexes were resolved via the combined actions of electrophoretic migration and electro-osmotic flow along a buffer-filled, uncoated fused-silica capillary. The I-CZE fractions collected from both CymMV- and ORSV-antibody complex peaks, as well as the RNA purified from them, retained their infectivity upon inoculation onto Chenopodium quinoa. I-CZE assays were able to detect as little as 10 fg each of both CymMV and ORSV in their purified forms as well as in the crude saps of infected N. benthamiana and Oncidium orchid. As multiple samples can be analyzed rapidly, I-CZE offers an ideal diagnostic technique for routine mass-indexing programs such as virus-free certification, breeding for virus-resistant cultivars, plant quarantine, and germ plasm screening. This is the first report of the application of I-CZE for the detection of plant viruses.
摘要 免疫毛细管区带电泳（I-CZE）是一种将血清学检测的特异性与毛细管区带电泳的灵敏度、快速性和自动化检测相结合的技术。在纯化形式以及感染的烟草和文心兰叶片粗提物中检测到了蝴蝶兰 mosaic 马铃薯 Y 病毒（CymMV）和文心兰环斑烟草花叶病毒（ORSV）。两种兰花病毒-抗体复合物通过电泳迁移和缓冲液填充的未涂层熔融硅毛细管中的电渗流的联合作用得以分离。从 CymMV-和 ORSV-抗体复合物峰中收集的 I-CZE 馏分以及从中纯化的 RNA 在接种藜麦后仍保留其感染性。I-CZE 分析能够检测到低至 10 fg 的纯化形式以及感染的烟草和文心兰叶片粗提物中的 CymMV 和 ORSV。由于可以快速分析多个样本，因此 I-CZE 为常规大规模指数检测程序（例如无病毒认证、抗病毒品种选育、植物检疫和种质筛选）提供了理想的诊断技术。这是首次应用 I-CZE 检测植物病毒的报道。
