Tang Ya-bing, Xing Da, Zhu De-bin, Liu Jin-feng
MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, South China Normal University, Guangzhou, China.
Anal Chim Acta. 2007 Jan 23;582(2):275-80. doi: 10.1016/j.aca.2006.09.021. Epub 2006 Sep 16.
Recently, we have reported an electrochemiluminescence polymerase chain reaction (ECL-PCR) method for detection of genetically modified organisms. The ECL-PCR method was further improved in the current study by introducing a multi-purpose nucleic acid sequence that was specific to the tris(bipyridine) ruthenium (TBR) labeled probe, into the 5' terminal of the primers. The method was applied to detect plant viruses. Conserved sequence of the plant viruses was amplified by PCR. The product was hybridized with a biotin labeled probe and a TBR labeled probe. The hybridization product was separated by streptavidin-coated magnetic beads, and detected by measuring the ECL signals of the TBR labeled. Under the optimized conditions, the experiment results show that the detection limit is 50 fmol of PCR products, and the signal-to-noise ratio is in excess of 14.6. The method was used to detect banana streak virus, banana bunchy top virus, and papaya leaf curl virus. The experiment results show that this method could reliably identity viruses infected plant samples. The improved ECL-PCR approach has higher sensitivity and lower cost than previous approach. It can effectively detect the plant viruses with simplicity, stability, and high sensitivity.
最近，我们报道了一种用于检测转基因生物的电化学发光聚合酶链反应（ECL-PCR）方法。在当前研究中，通过将对三联吡啶钌（TBR）标记探针特异的多用途核酸序列引入引物的5'末端，对ECL-PCR方法进行了进一步改进。该方法应用于检测植物病毒。通过PCR扩增植物病毒的保守序列。产物与生物素标记探针和TBR标记探针杂交。杂交产物用链霉亲和素包被的磁珠分离，并通过测量TBR标记的电化学发光信号进行检测。在优化条件下，实验结果表明检测限为50 fmol的PCR产物，信噪比超过14.6。该方法用于检测香蕉条纹病毒、香蕉束顶病毒和木瓜叶卷曲病毒。实验结果表明，该方法能够可靠地鉴定病毒感染的植物样品。改进后的ECL-PCR方法比以前的方法具有更高的灵敏度和更低的成本。它可以简单、稳定且高灵敏度地有效检测植物病毒。
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