论文部分内容阅读
根据5种病毒小西葫芦黄花叶病毒(Zucchini yellow mosaic virus,ZYMV)、西瓜花叶病毒(Watermelon mosaic virus,WMV)、烟草花叶病毒(Tobacco mosaic virus,TMV)、南瓜花叶病毒(Squash mosaic virus,SqMV)和黄瓜花叶病毒(Cucumber mosai cvirus,CMV)的核苷酸保守区序列,设计特异性引物对,从影响多重RT-PCR(mRT-PCR)扩增的引物浓度、Mg2+浓度、TaqDNA聚合酶浓度、dNTPs浓度、退火温度等方面进行反应体系的优化,建立了一种能够同时检测ZYMV、WMV、TMV、SqMV和CMV的多重RT-PCR技术体系,并进行了实际应用。在一个体系中对上述5种病毒复合侵染的西瓜材料进行多重RT-PCR扩增,得到与试验设计相符的5条特异性条带,依次是542、485、410、354和293bp。该体系实现了对侵染西瓜的5种病毒的同时检测,极大地提高了检测效率,降低了检测成本,体现了多重RT-PCR的优越性。
According to five kinds of viruses, Zucchini yellow mosaic virus (ZYMV), Watermelon mosaic virus (WMV), Tobacco mosaic virus (TMV), Squash mosaic virus , SqMV) and Cucumber mosai cvirus (CMV) were designed according to the nucleotide sequences of the conserved regions of the primers. Polymerase chain reaction, polymerase, dNTPs concentration and annealing temperature. A multi-RT-PCR system for simultaneous detection of ZYMV, WMV, TMV, SqMV and CMV was established and applied. In a system of five kinds of virus complex infection of watermelon material by multiple RT-PCR amplification, and experimental design consistent with the five specific bands, followed by 542,485,410,354 and 293bp. The system realizes the simultaneous detection of 5 viruses that infect watermelon, greatly improves the detection efficiency, reduces the detection cost and reflects the superiority of multiplex RT-PCR.