| 柏自琴.仙人指X病毒实时荧光定量PCR检测体系的构建及在火龙果上的应用[J].中国南方果树,2026,55(3): |
| 仙人指X病毒实时荧光定量PCR检测体系的构建及在火龙果上的应用 |
| Establishment of real-time fluorescence quantitative PCR detection system for schlumbergera virus X and its application in pitaya |
| 投稿时间：2026-04-23 修订日期：2026-06-05 |
| DOI： |
| 中文关键词: 仙人指X病毒 火龙果 实时荧光定量PCR 检测 |
| 英文关键词:schlumbergera virus X pitaya real-time fluorescence quantitative PCR detection |
| 基金项目: |
|
| 摘要点击次数: 119 |
| 全文下载次数: 2 |
| 中文摘要: |
| 【目的】仙人指X病毒（schlumbergera virus X，SchVX）的流行对火龙果种植业造成显著危害，为实现对该病毒的定量检测，特开展此工作。【方法】本研究基于多个SchVX分离株外壳蛋白的保守区域，设计特异性引物组合SchVX-F/-R，开发了SYBR Green I实时荧光定量PCR检测体系。【结果】在引物工作浓度150 nmol/L，退火条件61℃下，获得96.95%的扩增效率与0.999 6的线性相关系数，最低检测限达8×102拷贝/μL，较常规PCR灵敏度提升两个数量级。病毒组织分布分析表明，病毒在火龙果花瓣中丰度最高，含量依次递减为老枝＞花萼＞根系＞嫩枝＞花药。在黔南地区田间40份样品的检测中，该方法成功检出35例阳性样本，阳性检出率达87.5%。【结论】实验数据证实该方法兼具良好的灵敏性与准确性，为SchVX的早期诊断及定量分析提供了有效的分子检测工具。 |
| 英文摘要: |
| 【Objective】The prevalence of schlumbergera virus X (SchVX) poses a significant threat to the pitaya (dragon fruit) industry. This study aimed to establish a quantitative detection method for SchVX.【Methods】A specific primer pair, SchVX-F/-R, was designed based on the conserved region of the coat protein of multiple SchVX isolates, and a SYBR Green I real-time quantitative PCR (qPCR) detection system was developed.【Results】Under optimized conditions with a primer concentration of 150 nmol/L and an annealing temperature of 61℃, the assay achieved an amplification efficiency of 96.95% and a linear correlation coefficient (R2) of 0.999 6.
This study developed a SYBR Green I real-time fluorescence quantitative PCR (RT-qPCR) detection system based on the conserved regions of the coat protein of multiple SchVX isolates. Using a specific primer combination SchVX-F/-R (working concentration 150 nmol/L, annealing condition 61℃), a linear correlation coefficient of 0.999 6 was obtained with an amplification efficiency of 96.95%. The lowest detection limit was 8×102 copies/μL DNA, which was two orders of magnitude higher than conventional PCR sensitivity. The analysis of virus tissue distribution shows that the virus is most abundant in the petals of dragon fruit, and its content decreases in the order of old branches>calyxes>root>tender branches>anthers. In the testing of 40 samples in the fields of south of Guizhou Province, this method successfully detected 35 positive samples, with a positive detection rate of 87.5%.【Conclusion】The experimental data confirms that this method has both good sensitivity and accuracy, providing an effective molecular detection tool for the early diagnosis and quantitative analysis of SchVX. |
| 查看/发表评论 下载PDF阅读器 |
| 关闭 |
