Detection of Cymbidium Mosaic Potexvirus and Odontoglossum Ringspot Tobamovirus Using Immuno-Capillary Zone Electrophoresis

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.

A Basic Approach Towards the Development of Bioelectric Bacterial Biosensors for the Detection of Plant Viruses

“…2008). Other detection technologies include capillary zone electrophoresis (Eun and Wong 1999), liquid chromatography and matrix‐assisted desorption‐ionization (MALDI)‐mass spectrometry, quartz crystal microbalance‐based nucleic acid biosensor and nucleic acid sequence‐based amplification (NASBA) (Wong 2002). Eun et al.…”

Performances and application of antisera produced by recombinant capsid proteins of Cymbidium mosaic virus and Odontoglossum ringspot virus

“…tion methods based on targeting viral nucleic acids [such as reverse transcription-polymerase chain reaction (RT-PCR), hybridization and molecular beacon] (Lim et al 1993;Hu and Wong 1998;Seoh et al 1998;Lee and Chang 2006;Sherpa et al 2006), viral capsid proteins (CP) [such as enzyme-linked immunosorbent assay (ELISA), quartz crystal microbalance (QCM) immunosensors, immuno-capillary zone electrophoresis (I-CZE), liquid chromatography/mass spectrometry (LC/MS) and matrix-assisted laser desorption-ionization (MALDI) mass spectrometry] (Arunasalam and Pearson 1989;Wong and Chng 1993;Eun and Wong 1999;Tan et al 2000;Eun et al 2002) or both [such as immunocapture-PCR (IC-PCR)] (Barry et al 1996) were developed. Among these detection methods, ELISA is cost-effective and shows satisfying detection sensitivity (1-10 ng ml −1 ; Hull 2002).…”

Orchid Biology: Reviews and Perspectives, VIII

“…Immuno-capillary zone electrophoresis (CZE) is able to detect as little as 10 fg each of both CymMV and ORSV in their crude saps ofinfected Oncidium orchid ( Eun and Wong, 1999). Immuno-capillary zone electrophoresis.…”