中文摘要：玉米褪绿斑驳病毒会导致玉米坏死病并可通过感染玉米种子进行传播，该病毒的检测一直是一个挑战，因此，本文首先对新型检测方法的不对称RT-PCR引物比、NaCl浓度以及特异性和敏感性进行了优化，并据此开发了一种以未修饰金纳米粒子为比色探针，采用不对称聚合酶链式反应可视化检测玉米褪绿斑驳病毒的方法。使用该方法，在不借助任何昂贵的仪器和生化试剂的情况下，即可通过肉眼观察到浓度在23.9 pg /μL以下的的玉米褪绿斑驳病毒RNA。该方法具有高种属特异性、简单、低成本、可视化等优点，在中小型企业和出入境检疫局对玉米种子和植物组织中玉米褪绿斑驳病毒检测方面具有较大的应用潜力。
外文摘要：Maize chlorotic mottle virus causes corn lethal necrosis disease and can transmit via infected maize seeds. It remains a challenge to detect this virus to prevent its introduction and infection and spread transmission fields. For this purpose, visual detection of Maize chlorotic mottle virus was investigated in cooperation with a asymmetric polymerase chain reaction and unmodified gold nanoparticles (AuNPs). Detection relies on the fact that the dispersed AuNPs solution is red due to the intense surface plasmon absorption band at 520 nm. The color changed to grey after saline induction; single-stranded DNA (ssDNA) electrostatically adsorbed onto naked AuNPs stabilizes them against salt-induced aggregation, whereas the AuNPs solution in the presence of ssDNA of asymmetric RT-PCR target products of Maize chlorotic mottle virus remains red even after saline induction. A remarkable color change from red to grey immediately occurred in absence of ssDNA amplified from Maize chlorotic mottle virus with asymmetric RT-PCR. After optimization of the asymmetric RT-PCR primer ratios, concentration of NaCl, and evaluation of specificity and sensitivity of a novel assay, visual detection of Maize chlorotic mottle virus using unmodified AuNPs and asymmetric RT-PCR has been developed with simple preparation of samples in our study. Through this assay, as low as 23.9 pg mu L-1 of RNA of Maize chlorotic mottle virus was detected visually by the naked eye without the need for any sophisticated, expensive instrumentation and biochemical reagents. The results indicate that this assay is highly species-specific, simple, low-cost, and visible for easy detection of Maize chlorotic mottle virus in plant tissues. Therefore, visual detection of Maize chlorotic mottle virus is a potentially useful tool for middle-or small-scale corporations and entry-exit inspection and quarantine bureaux to detect Maize chlorotic mottle virus in maize seeds or plant tissues.
 
外文关键词：SINGLE-STRANDED-DNA; FRANKLINIELLA-WILLIAMSI; AMPLIFICATION; SILVER; TRANSMISSION; BIOSENSOR; MODE
作者：Wang, Lin; Liu, Zhanmin; Xia, Xueying; Huang, Junyi
作者单位：Shanghai Univ, Sch Life Sci, 333 Nanchen Rd, Shanghai 200444, Peoples R China
期刊名称：ANALYTICAL METHODS
期刊影响因子：1.915
出版年份：2016
出版刊次：8
点击下载：未修饰的金纳米粒子比色探针与不对称聚合酶链反应联用可视化检测玉米褪绿斑驳病毒