Detection of cucumber mosaic cucumovirus in weed species: a cautionary report on nonspecific reactions in ELISA

E. Sikora, M. Andrianifahanana, John F. Murphy
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引用次数: 7

Abstract

The reliability of enzyme-linked immunosorbent assay (ELISA) to detect cucumber mosaic cucumovirus (CMV) coat protein (CP) in three weed species (Taraxacum officinale, Gnaphalium obtusifolium, and Conyza canadensis) was compared with other detection methods, including Western blot analysis and a bioassay. Analysis of extracts prepared from these species by double antibody sandwich (DAS) ELISA yielded high absorbance values (A4O5nm) that were indicative of CMV concentrations of purified CMV standards of greater than 50 mg/mL. However, results from an ELISA procedure designed such that no viral antigen should be detected suggested that the strong ELISA absorbance values were nonspecific reactions with material(s) in the weed extracts. These findings were substantiated when no CMV CP was detected by Western blot analysis or infectious CMV was detected in bioassays involving inoculation of extracts onto an indicator host (Cucurbita pepo ‘Crookneck’ (squash)). In both DAS ELISA and indirect ELISA procedures, t...
黄瓜花叶黄瓜病毒在杂草中的检测:酶联免疫吸附法非特异性反应的警示报告
比较了酶联免疫吸附法(ELISA)检测三种杂草(Taraxacum officinale、Gnaphalium obtusiolium和Conyza canadensis)中黄瓜花叶病毒(CMV)外膜蛋白(CP)的可靠性。双抗体夹心(DAS)酶联免疫吸附法(ELISA)对这些物种的提取物进行分析,获得了高吸光度值(A4O5nm),表明纯化的CMV标准物的CMV浓度大于50 mg/mL。然而,ELISA程序设计的结果表明,不应检测到病毒抗原,强ELISA吸光度值是与杂草提取物中的物质的非特异性反应。当Western blot分析未检测到CMV CP或将提取物接种到指示宿主(Cucurbita pepo ' Crookneck '(南瓜))上的生物测定中检测到感染性CMV时,这些发现得到了证实。在DAS ELISA和间接ELISA方法中,t…
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