Development and Application of a Quantitative RT-PCR Approach for Quantification of T3 Genotype of Citrus tristeza virus
Development and Application of a Quantitative RT-PCR Approach for Quantification of T3 Genotype of Citrus tristeza virus
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柑橘 tristeza 病毒 T3 基因型定量 RT-PCR 方法的开发和应用
DOI：
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发表时间：
2015
期刊：
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通讯作者：
Tao Zhen
中科院分区：
文献类型：
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作者：
Tao Zhen
根据柑橘衰退病毒（Citrus tristeza virus，CTV）ORF 1 a基因的保守序列，设计引物T3-4F/R，建立了CTV ORF 1 a基因的定量RT-PCR方法。结果表明，该方法的检测效率比常规PCR方法高出至少百倍。两条标准曲线线性关系良好（R2 = 0.992）。扩增效率为97.1%。三次重复检测结果表明，批内、批间变异系数均在2.94%以内，是一种重复性较好的CTV检测方法。田间样品中T3基因型含量差异显著，最高可达最低的1 250倍。该方法可准确地检测出T3基因型，并可用于研究T3基因型的变异。
This study established a quantitative RT-PCR method with primers T3-4F/R based on the conserved nucleotide sequence of Citrus tristeza virus(CTV)gene ORF1 a. The results showed that the method was at least hundred times higher than that with conventional PCR. A good linear correlation(R2 = 0.992)obtained from two standard curve of c RNA. The amplification efficiency was 97.1%. Three-time repeats revealed that the coefficients of variation between the intra- and inter-assay were both within 2.94%,indicating a reliating reproducibility detection method to CTV. There was noticeable differences among the content of T3 genotype in field samples,the highest content can reach to 1 250 times the lowest. The method was used for accurate determination of T3 genotype in the plant,and could be used to study the variation of T3 genotype.