State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Guiyang 550025, China.
Molecules. 2012 Jun 5;17(6):6886-900. doi: 10.3390/molecules17066886.
Southern rice black-streaked dwarf virus (SRBSDV), a new virus from Fiji, has seriously damaged rice crops in southern China and northern Vietnam in recent years. This virus is difficult to diagnose in the early stages of infection, and is very destructive at the late stage. In the present study, a dot immunobinding assay (DIBA) that has a high sensitivity for diagnosing SRBSDV was developed. Two kinds of treatment for the DIBA were evaluated to determine the most effective one for removing chlorophyll interferences via rice extraction. The first included several reagents to remove chlorophyll, namely, the alkaline reagents like magnesium oxide and alumina oxide, the adsorbent reagents like activated carbon and bentonite, as well as the extraction agent acetone. The second and third treatments, which were used to remove chlorophyll in blot membrane-nitrocellulose and polyvinylidene fluoride (PVDF), included several organic solvents containing methanol, ethanol, acetone, ethyl acetate, and diethyl ether. The results showed that activated carbon and methanol yielded the best contrasting purple color for the infected samples by decreasing the chlorophyll content.
南方水稻黑条矮缩病毒（SRBSDV）是一种来自斐济的新型病毒，近年来严重破坏了中国南方和越南北部的水稻作物。该病毒在感染初期难以诊断，在后期具有很强的破坏性。本研究开发了一种具有高灵敏度的用于诊断 SRBSDV 的斑点免疫结合测定法（DIBA）。评估了 DIBA 的两种处理方法，以确定通过水稻提取去除叶绿素干扰最有效的方法。第一种方法包括几种去除叶绿素的试剂，如氧化镁和氧化铝等碱性试剂、活性炭和膨润土等吸附剂试剂以及丙酮等提取剂。第二种和第三种处理方法，用于去除斑点膜-硝酸纤维素和聚偏二氟乙烯（PVDF）中的叶绿素，包括几种含有甲醇、乙醇、丙酮、乙酸乙酯和乙醚的有机溶剂。结果表明，通过降低叶绿素含量，活性炭和甲醇为受感染的样品产生了最佳的对比紫色，效果最好。
