柑橘衰退病毒RB和VT基因型实时定量PCR检测方法的建立和应用

为定量检测柑橘衰退病毒（citrus tristeza virus, CTV）RB、VT基因型的含量，本研究以RB基因型的p33基因、VT基因型的ORF1a基因为靶标，建立了RB、VT基因型特异性的实时定量PCR方法，该方法灵敏度为普通PCR的1000倍；RB、VT基因型标准曲线的相关性系数分别为0.9991、0.9954，扩增效率分别为99.85%、102.05%；两种方法组内和组间变异系数均小于2.07%，重复性良好。对田间样品检测发现RB、VT基因型含量差异较大。本研究建立的方法特异性强，灵敏度高，适用于田间样品检测。

Development and Application of a Quantitative PCR Approach for Quantification of RB or VT Genotype of Citrus tristeza virus

In order to quantitatively detect the RB and VT genotypes of citrus tristeza virus (CTV), a genotype-specific real-time quantitative PCR method was established by targeting the p33 gene of RB genotype and the ORF1a gene of VT genotype. The sensitivity of this method was 1000 times higher than that of conventional PCR. The correlation coefficients of the standard curve of RB and VT genotypes were 0.9991 and 0.9954, respectively, and the amplification efficiencies were 99.85% and 102.05%, respectively. The intra - and inter-group coefficients of variation were both within 2.07%, respectively, indicating that both methods were stable. The contents of RB or VT genotypes in the field samples were significantly different. The method established in this study has high specificity and sensitivity, and is suitable for the detection of samples in the field.