Reversetranscription, real-time PCR assay for detection of Toscana virus
Reversetranscription, real-time PCR assay for detection of Toscana virus
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DOI：
10.1016/j.jcv.2007.05.003
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发表时间：
2007-08-01
影响因子：
8.8
通讯作者：
Tenorio, Antonio
中科院分区：
文献类型：
--
作者：
Perez-Ruiz, Mercedes;Collao, Ximena;Tenorio, Antonio
背景：节肢动物传播的托斯卡纳病毒是地中海盆地急性神经系统感染的常见原因。最近，西班牙报道了一种与意大利原型高度不同的新血统。目的：建立一种检测两种托斯卡纳病毒基因型的反转录实时PCR检测方法。实时PCR采用TaqMan (R)探针和内控来识别假阴性结果。研究设计：选择托斯卡纳病毒两个已知谱系的保守区域，位于其基因组小片段的3'端，设计引物和探针。结果：检测Toscana病毒的灵敏度为0.0158 TICD50，相当于7份cDNA。没有其他的白蛉病毒或RNA病毒被特异的实时PCR扩增。结论：该方法灵敏、可靠，可用于本地和（或）输入性托斯卡纳病毒感染疑似病例的诊断。(c) 2007 Elsevier B.V.版权所有
Background: The arthropod-borne Toscana virus is a common cause of acute neurological infection in the Mediterranean basin. Recently, a new lineage, highly divergent from the Italian prototype, has been reported in Spain.Objective: We describe a reversetranscription, real-time PCR assay for detection of both Toscana virus genotypes. The real-time PCR uses a TaqMan (R) probe and an internal control to identify false negative results.Study design: A conserved region of the two known lineages of Toscana virus, located at the 3' end of the small segment of their genomes, was chosen to design both the primers and the probe.Results: The sensitivity of the assay was 0.0158 TICD50 per reaction of Toscana virus, equivalent to seven copies of cDNA. No other phleboviruses or RNA viruses were amplified by this specific real-time PCR.Conclusions: The assay seems to be sensitive, reliable and easy to be applied in the diagnosis of autochthonous and/or imported suspected cases of Toscana virus infection. (c) 2007 Elsevier B.V. All rights reserved.