对虾桃拉病毒（TSV）RT—PCR快速诊断技术的研究

• 作 者: 徐丽美 吴成林 邱名毅 王海贵 李丹萍 杨丰
• 机 构: 国家海洋局第三海洋研究所海洋生物遗传资源重点实验室,国家海洋局第三海洋研究所海洋生物遗传资源重点实验室,海南省水产养殖病害防治中心,海南省水产养殖病害防治中心,海南省水产养殖病害防治中心,国家海洋局第
• 出 处: 厦门大学学报(自然科学版)
• 发表日期: 2005年02期
• 关键词: 对虾桃拉病毒 TSV 凡纳对虾 P.vannamei RT-PCR 快速诊断技术 RNA病毒 Taura Syndrome Virus(TSV) RT-PCR

对虾桃拉病毒(Taura Syndrome Virus,TSV)最早于1992 年在凡纳对虾(Penaeus vannamei)体内发现,此病毒是对虾养殖业危害较严重的病毒之一.将该病毒基因组的一段序列克隆至转录载体 pSP64(PolyA)上,经体外转录、磁珠法分离、纯化获得了人工的TSV RNA模板;用定量的TSV RNA阳性对照模板进行RT PCR条件优化,灵敏度检测以及样品制备方法等试验,结果表明:RT PCR检测的灵敏度可达到0.1 fg,相当于 100 个病毒粒子;所制备的样品对病毒 RNA的逆转录及扩增无抑制,适合于对虾桃拉病毒快速检测.

Taura Syndrome Virus (TSV) was firstly found in Penaeus vannamei in 1992. It was one of the most harmful viruses in shrimp farming.A sequence of the virus genome was cloned into the transcript The artificial TSV RNA template was obtained by in vitro transcription and magnetic bead separation on the pSP64 (PolyA) vector. The optimized TSV RNA positive control template was used to optimize the RT PCR conditions, the sensitivity test and the sample preparation method. The results showed that : The sensitivity of RT PCR detection can reach 0.1 fg, which is equivalent to 100 virus particles. The prepared sample has no inhibition on the reverse transcription and amplification of viral RNA, and is suitable for the rapid detection of Tahu Tara virus.