XIE Lixue,CAI Wei,ZHENG Shan,et al.Rapid Detection of Blueberry Shock Virus by RT-PCR[J].Northern Horticulture,2015,39(20):98-101.[doi:10.11937/bfyy.201520024]
蓝莓休克病毒的RT-PCR快速检测方法建立
- Title:
- Rapid Detection of Blueberry Shock Virus by RT-PCR
- Keywords:
- blueberry shock virus; RT-PCR; rapid detection
- 文献标志码:
- A
- 摘要:
- 以蓝莓叶片为试材，提取病毒RNA，根据已报道的蓝莓休克病毒（Blueberry shock virus，BlShV）外壳蛋白基因序列设计特异性引物，建立RT-PCR快速检测方法，并对采集的20份疑似病例进行检测。结果表明：建立的RT-PCR方法具有良好的特异性和较高的灵敏度。仅从感染BlShV的样品中扩增出与预期大小相符的特异性目的片段，而从其它蓝莓病毒及健康样品中未扩增到目的片段；该方法能检测到RNA原液的10-4倍稀释液，灵敏度较高。疑似病例检测结果与ELISA验证结果完全相符。试验表明，建立的RT-PCR方法能够用于BlShV的现地检测。
- Abstract:
- RT-PCR method was established for rapid detection of Blueberry shock virus (BlShV).A set of specific primers were designed based on the published nucleotide sequence of BlShV coat protein gene.The viral RNA extracted from infected blueberry sample was used as template.Twenty suspected blueberry samples were detected by using the established method.The results showed that RT-PCR method had good specificity and high sensitivity.The method could amplify target fragment from BlShV-infected sample，while not from other blueberry virus infected sample and healthy sample.The sensitivity of RT-PCR method was 10-4 RNA.The detection result of suspected blueberry samples was consistent with ELISA，which indicated the RT-PCR method can be actually used for detection of BlShV.
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备注/Memo
第一作者简介：谢丽雪（1981-），女，福建厦门人，硕士，助理研究员，研究方向为果树遗传育种及病害。E-mail：sarah614510@126.com.责任作者：李韬（1969-），男，本科，副研究员，研究方向为果树遗传育种及病害。E-mail：leetao06@163.com.基金项目：福建省公益类科研院所专项资助项目（2015R110101813）;福建省种业创新与产业化工程资助项目（2014S1477-22）；福州市科技计划资助项目（2013-N-54）；福建省农业科学院导师制基金资助项目（2013DQB-9）。