Development and application of a universal and simplified multiplex RT-PCR assay to detect five potato viruses

文献类型： 外文期刊

第一作者： Wen, Jingzhi

作者： Wen, Jingzhi;Zhang, Wei;Fan, Guoquan;Gao, Yanling;Bai, Yanju;Qiu, Cailing;Zhang, Shu;Shen, Yu;Zhang, Zhixiang;Meng, Xianxin

作者机构：

关键词： Potato;PVX;PVY;PLRV;PVM;PVA;Multiplex RT-PCR;Detection

期刊名称：JOURNAL OF GENERAL PLANT PATHOLOGY （ 影响因子：1.449； 五年影响因子：1.416 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： Potato virus X (PVX), Potato virus Y (PVY), Potato leafroll virus (PLRV), Potato virus M (PVM), and Potato virus A (PVA) commonly cause mixed infections and major economic losses in potato crops, so a multiplex reverse transcription polymerase chain reaction (RT-PCR) assay has been greatly needed to detect all five viruses simultaneously. Previously published primers were evaluated, and five pairs of highly specific primers were selected for use. The quintuplex RT-PCR assay was optimized for primer concentration, annealing temperature, number of PCR cycles, and extension time. Finally, the quintuplex RT-PCR assay was successfully established. The amplified target fragments of 711 base pairs (bp) for PVX, 447 bp for PVY, 336 bp for PLRV, 520 bp for PVM and 273 bp for PVA, and the viral origins of the PCR products were confirmed by sequencing. This system can be easily customized for simplex, duplex, triplex, tetraplex, and quintuplex RT-PCRs by changing only the primer pairs while keeping the final reaction volume and the concentrations of other reagents constant to detect any combination of PVX, PVY, PLRV, PVM, and PVA viruses. In this way, the detection systems can be customized to the characteristics of viral infections specific to China's potato-producing regions for excellent detection efficiency and meeting the needs of potato producers. The detection limit of this assay was up to 10(-3) dilution in tetraplex and quintuplex RT-PCR, and even higher in simplex, duplex, and triplex RT-PCR. The efficiency of the quintuplex RT-PCR was compared with that of a DAS-enzyme-linked immunosorbent assay to detect the five viruses in 320 field potato samples, 195 in vitro plantlets and pooled potato samples. This new multiplex RT-PCR method is not only effective but also faster, cheaper, more sensitive, and more reliable than DAS-ELISA. It is especially suitable for the testing of in vitro plantlets and pooled samples of postharvest seed tubers.

分类号： S432.1

  • 相关文献

[1]Impact of aphid alarm pheromone release on virus transmission efficiency: When pest control strategy could induce higher virus dispersion. Liu, Ying-Jie,Liu, Yong,Lin, Fang-Jing,Bosquee, Emilie,Francis, Frederic,Chen, Ju-Lian.

[2]Development and Validation of a Multiplex Reverse Transcription PCR Assay for Simultaneous Detection of Three Papaya Viruses. Tuo, Decai,Shen, Wentao,Yang, Yong,Yan, Pu,Li, Xiaoying,Zhou, Peng,Tuo, Decai. 2014

[3]A multiplex RT-PCR assay for simultaneous detection of four viruses from sweet cherry. Zong, Xiaojuan,Wang, Wenwen,Wei, Hairong,Wang, Jiawei,Chen, Xin,Xu, Li,Zhu, Dongzi,Tan, Yue,Liu, Qingzhong.

[4]A Novel Recombined Potato virus Y Isolate in China. Han, Shuxin,Gao, Yanling,Zhang, Wei,Zhang, Junhua,Gao, Yanling,Fan, Guoquan,Zhang, Wei,Qiu, Cailing,Zhang, Shu,Bai, Yanju,Spetz, Carl. 2017

[5]Establishment of a multiplex RT-PCR assay to detect different lineages of swine H1 and H3 influenza A viruses. Fu, Guanghua,Liu, Mengda,Zeng, Wenshu,Pu, Juan,Bi, Yuhai,Liu, Jinhua,Fu, Guanghua,Ma, Guangpeng,Liu, Jinhua. 2010

[6]Multiplex RT-PCR detection and distribution of four apple viruses in China. Ji, Z.,Zhao, X.,Duan, H.,Hui, T.,Wang, S.,Wang, Y.,Cao, K.,Ji, Z.. 2013

[7]Rapid discrimination of H5 and H9 subtypes of avian influenza viruses and Newcastle disease virus by multiplex RT-PCR. Chen, Hao-tai,Zhang, Jie,Sun, De-hui,Zhang, Ji-le,Cai, Xue-peng,Liu, Xiang-tao,Ding, Yao-zhong,Ma, Li-na,Yang, Sheng-hai,Jin, Lei,Liu, Yongsheng. 2008

[8]Simultaneous detection of wheat dwarf virus, northern cereal mosaic virus, barley yellow striate mosaic virus and rice black-streaked dwarf virus in wheat by multiplex RT-PCR. Liu, Yan,Liu, Wenwen,Wang, Xifeng,Zhang, Peipei,Massart, Sebastien.

[9]A two-step multiplex RT-PCR method for simultaneous detection of soil-borne wheat mosaic virus and wheat spindle streak mosaic virus from France. Gitton, F,Diao, A,Ducrot, O,Antoniw, JF,Adams, MJ,Maraite, H. 1999

[10]A one-step multiplex RT-PCR assay for simultaneous detection of four viruses that infect peach. Zhao, Z.,Jiang, D.,Li, S.,Yu, Y.,Jiang, D.,Wu, Z.,Zhao, Z.. 2013

[11]A Multiplex RT-PCR Assay to Detect and Discriminate Porcine Reproductive and Respiratory Syndrome Viruses in Clinical Specimens. Yang, Keli,Tian, Yongxiang,Zhou, Danna,Duan, Zhengying,Guo, Rui,Liu, Zewen,Yuan, Fangyan,Liu, Wei. 2017

[12]Rely on science-technology value chain, strength the management of technological innovation strategic alliance of potato industry. Qu, Huijuan,Liu, Dinghui,Wang, Hong,Huang, Gang. 2011

[13]Construction of Reference Chromosome-Scale Pseudomolecules for Potato: Integrating the Potato Genome with Genetic and Physical Maps. Sharma, Sanjeev Kumar,McLean, Karen,Waugh, Robbie,Bryan, Glenn J.,Bolser, Daniel,Martin, David Michael Alan,de Boer, Jan,Visser, Richard G. F.,Bachem, Christian W. B.,Sonderkaer, Mads,Amoros, Walter,Munive, Susan,Simon, Reinhard,Bonierbale, Merideth,Carboni, Martin Federico,Feingold, Sergio E.,D'Ambrosio, Juan Martin,de la Cruz, German,Di Genova, Alex,Maass, Alejandro,Di Genova, Alex,Maass, Alejandro,Douches, David S.,Hamilton, John P.,Eguiluz, Maria,Guzman, Frank,Lozano, Roberto,Martinez, Diana,Ponce, Olga,Ramirez, Manuel,Torres, Yerisf,Orjeda, Gisella,Guo, Xiao,Li, Guangcun,Hackett, Christine A.,Li, Ying,Thomson, Susan J.,Zhang, Zhonghua,Huang, Sanwen,Mejia, Nilo,Nagy, Istvan,Milbourne, Dan,Jacobs, Jeanne M. E.,Jacobs, Jeanne M. E.,Veilleux, Richard E.. 2013

[14]Soil nutrient loss due to tuber crop harvesting and its environmental impact in the North China Plain. Yu Han-qing,Li Yong,Zhou Na,Li Xiao-yu,Chappell, Adrian,Poesen, Jean. 2016

[15]Potato Spectrum and the Digital Image Feature Parameters on the Response of the Nitrogen Level and Its Application. Zheng Shun-lin,Wan Nian-xin,Zhao Ting-ting,Yuan Ji-chao,Zheng Shun-lin,Yuan Ji-chao,He Wei,Hu Jian-jun. 2016

[16]Potential and actual yields of potato at different elevations and in different seasons in subtropical Southwest China. Struik, PC,Wang, J,Zhang, X. 1998

[17]Effects of Exogenous Chitosan on Physiological Characteristics of Potato Seedlings Under Drought Stress and Rehydration. Jiao, Zhili,Li, Yong,Lu, Dianqiu,Li, Juanjuan,Xu, Xiaoyan,Wang, Jingying,Li, Hui. 2012

[18]Late Blight Management in China. Hu, Tongle,Wang, Fengyi,He, Wei,Xie, Kaiyun. 2009

[19]Cell division and cell enlargement during potato tuber formation. Xu, X,Vreugdenhil, D,van Lammeren, AAM. 1998

[20]Planting time and seed density effects on potato in subtropical China. Struik, PC,He, Q,Zhang, X. 1998

作者其他论文 更多>>

微信小程序