Rowhani Adib, Biardi Lenka, Routh Geoffrey, Daubert Steve D, Golino Deborah A
Plant Pathology Specialist.
Postgraduate Researcher.
Plant Dis. 1998 Aug;82(8):880-884. doi: 10.1094/PDIS.1998.82.8.880.
Diagnostic methods employing the polymerase chain reaction (PCR) provide the most sensitive means currently available for detecting viruses in woody plants. A new technique has been tested that does not rely on gel electrophoresis or molecular hybridization to detect virus-specific PCR products. This colorimetric method for detection of PCR products from woody plants was demonstrated to be at least as sensitive as gel analysis. When combined with immunocapture of virions from plant sap, colorimetric detection provides a means to apply PCR technology to a large number of samples. Here, we report on the use of this technique for detection and quantitation of a walnut isolate of cherry leafroll virus (CLRV-W), citrus tristeza virus (CTV), prune dwarf virus (PDV), prunus necrotic ringspot virus (PNRSV), and tomato ringspot virus (ToRSV) in woody and herbaceous plants. For purified virus preparations, detection limits ranged from 100 pg/ml to 100 ag/ml. We also describe the colorimetric PCR detection of CTV in pooled samples.
采用聚合酶链反应（PCR）的诊断方法是目前检测木本植物中病毒最灵敏的手段。一种不依赖凝胶电泳或分子杂交来检测病毒特异性PCR产物的新技术已得到测试。这种用于检测木本植物PCR产物的比色法被证明至少与凝胶分析一样灵敏。当与从植物汁液中免疫捕获病毒粒子相结合时，比色检测提供了一种将PCR技术应用于大量样品的方法。在此，我们报告了该技术用于检测和定量木本和草本植物中核桃分离株的樱桃卷叶病毒（CLRV-W）、柑橘衰退病毒（CTV）、李矮缩病毒（PDV）、李坏死环斑病毒（PNRSV）和番茄环斑病毒（ToRSV）的情况。对于纯化的病毒制剂，检测限范围为100 pg/ml至100 ag/ml。我们还描述了在混合样品中CTV的比色PCR检测。
