葫芦科作物3种主要病毒的多重RT-PCR方法的建立Multiplex RT-PCR assay for simultaneous detection of three viruses of Zucchini
赵丽;古勤生;陈红运;史团省;田延平;胡京昂;
摘要(Abstract):
小西葫芦黄花叶病毒(Zucchini yellowmosaic virus,ZYMV)、西瓜花叶病毒(Watermelon mosaic virus,WMV)和黄瓜花叶病毒(Cucumber mosaic virus,CMV)是危害葫芦科作物最严重、最广泛的3种病毒。研究建立了以18SrRNA为内标检测ZYMV、CMV和WMV侵染葫芦科作物3种主要病毒的多重RT-PCR方法,并对影响多重RT-PCR扩增的退火温度、Taq聚合酶浓度、dNTPs浓度和镁离子浓度4种主要因素进行优化;对多重RT-PCR的灵敏性进行了测定,并与单重RT-PCR灵敏性进行比较。结果表明:最佳的退火温度、Taq聚合酶量、dNTPs浓度和Mg2+浓度分别为60.7℃、1.0U、0.8mmol/L和2.5mmol/L。多重RT-PCR各病毒的灵敏度与单重RT-PCR相同。
关键词(KeyWords): 西葫芦(Cucurbita moschata Duch);病毒病;多重RT-PCR
基金项目(Foundation): 河南省科技攻关项目(082102150012)
作者(Author): 赵丽;古勤生;陈红运;史团省;田延平;胡京昂;
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