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为实现田间土壤棉花黄萎病菌的早期检测,建立了土壤中棉花黄萎病菌的SYBR Green I荧光定量PCR检测方法。以含342bp PCR扩增产物的阳性质粒为参考,构建了标准曲线,并对该曲线的特异性、敏感性、可重复性进行了评价。结果表明,该方法具有快速、特异性强、敏感度高等特点。检测范围在3.8×103-3.8×108copies/μL之间有良好的线性关系,相关系数R2为0.996,扩增效率为101.5%,灵敏度比常规PCR方法高102倍。
In order to realize the early detection of Verticillium dahliae in field soil, SYBR Green I fluorescence quantitative PCR method was established for the determination of Verticillium dahliae in soil. A standard curve was constructed based on the positive plasmid containing 342 bp PCR amplification product. The specificity, sensitivity and repeatability of the curve were evaluated. The results show that this method is rapid, specific and sensitive. The detection range was good linearity between 3.8 × 103-3.8 × 108copies / μL, the correlation coefficient R2 was 0.996, the amplification efficiency was 101.5%, the sensitivity was 102 times higher than the conventional PCR method.