Multiplex RT-PCR Assay for the Simultaneous Detection and Identification of Five Sugarcane Viruses
Multiplex RT-PCR Assay for the Simultaneous Detection and Identification of Five Sugarcane Viruses
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同时检测和鉴定五种甘蔗病毒的多重 RT-PCR 测定
DOI：
10.1007/s12355-020-00805-2
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发表时间：
2020
期刊：
影响因子：
1.9
通讯作者：
Zhang Shu-Zhen
中科院分区：
文献类型：
--
作者：
Feng Xiao-Yan;Wang Wen-Zhi;Shen Lin-Bo;Wang Jun-Gang;Xiong Guo-Ru;Feng Cui-Lian;Zhao Ting-Ting;Zhang Shu-Zhen
甘蔗花叶病毒（SCMV）、高粱花叶病毒（SrMV）、甘蔗条纹花叶病毒（SCSMV）、甘蔗黄叶病毒（SCYLV）和甘蔗杆菌状病毒（SCBV）是世界范围内甘蔗感染的主要病毒。本研究提出了一种多重逆转录聚合酶链反应（RT-PCR）方法来同时检测和鉴定这些甘蔗病毒。结果显示，5对引物扩增出了各自的目标病毒，表明了检测的特异性。在检测SCMV和其他四种病毒时，该方法的灵敏度分别与单链RT-PCR相似或低10 - 100倍。在田间甘蔗样品中检测这5种病毒时，多重RT-PCR的检测结果与单多重RT-PCR的检测结果基本一致，表明前者相对可靠。多重RT-PCR可同时检测甘蔗中的SCMV、SrMV、SCSMV、SCYLV和SCBV，是一种适用、高效、实用的甘蔗病毒检测方法。
Sugarcane mosaic virus(SCMV),Sorghum mosaic virus(SrMV),Sugarcane streak mosaic virus(SCSMV),Sugarcane yellow leaf virus(SCYLV), andSugarcane bacilliform virus(SCBV) are the major viruses infecting sugarcane worldwide. This study proposed a multiplex reverse transcription–polymerase chain reaction (RT–PCR) assay for the simultaneous detection and identification of these sugarcane viruses. Results showed that five primer pairs amplified their respective target viruses, indicating the assay specificity. The sensitivity of the assay was similar to or 10–100-folds lower than that of singleplex RT–PCR for detecting SCMV or the four other viruses, respectively. When detecting the five viruses in field sugarcane samples, the detection results of multiplex RT–PCR were approximately identical to those of singleplex RT–PCR, thereby indicating the relative reliability of the former. The proposed multiplex RT–PCR can simultaneously detect SCMV, SrMV, SCSMV, SCYLV, and SCBV in sugarcane, making it a suitable, efficient, and practical detection method for sugarcane viruses.