Detection of all known filovirus species by reverse transcription-polymerase chain reaction using a primer set specific for the viral nucleoprotein gene.
Detection of all known filovirus species by reverse transcription-polymerase chain reaction using a primer set specific for the viral nucleoprotein gene.
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DOI：
10.1016/j.jviromet.2010.11.010
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发表时间：
2011-01
影响因子：
3.1
通讯作者：
Takada, Ayato
中科院分区：
文献类型：
--
作者：
Ogawa, Hirohito;Miyamoto, Hiroko;Ebihara, Hideki;Ito, Kimihito;Morikawa, Shigeru;Feldmann, Heinz;Takada, Ayato
丝状病毒、马尔堡病毒（MARV）和埃博拉病毒（EBOV）是严重出血热的病原体，在人类和非人类灵长类动物中死亡率很高。中非和亚洲部分地区出现了零星丝状病毒感染疫情。识别尚不清楚的自然宿主动物和流行病学调查是预防丝状病毒疾病爆发的当前挑战。目前已发现的丝状病毒种类包括 MARV 组中的一种和 EBOV 组中的五种，在不同种类之间发现了较大的遗传变异。因此，很难开发出一种单一的灵敏检测方法来检测所有丝状病毒种类，从而极大地推进流行地区的实验室诊断。在这项研究中，开发了一种针对丝状病毒核蛋白（NP）基因的高灵敏度通用 RT-PCR 检测方法。使用 NP 特异性引物组检测所有已知 MARV 和 EBOV 物种的基因组 RNA。此外，该RT-PCR程序在检测实验感染动物组织样本和感染患者血液样本中的病毒RNA方面的应用也得到了进一步验证。该测定将成为丝状病毒感染的诊断和流行病学研究的有用方法。
The filoviruses, Marburg virus (MARV) and Ebola virus (EBOV), are causative agents of severe hemorrhagic fever with high mortality rates in humans and non-human primates. Sporadic outbreaks of filovirus infection have occurred in Central Africa and parts of Asia. Identification of the natural reservoir animals that are unknown yet and epidemiological investigations are current challenges to forestall outbreaks of filovirus diseases. The filovirus species identified currently include one in the MARV group and five in the EBOV group, with large genetic variations found among the species. Therefore, it has been difficult to develop a single sensitive assay to detect all filovirus species, which would advance laboratory diagnosis greatly in endemic areas. In this study, a highly sensitive universal RT-PCR assay targeting the nucleo-protein (NP) gene of filoviruses was developed. The genomic RNAs of all known MARV and EBOV species were detected by using an NP-specific primer set. In addition, this RT-PCR procedure was verified further for its application to detect viral RNAs in tissue samples of animals infected experimentally and blood specimens of infected patients. This assay will be a useful method for diagnostics and epidemiological studies of filovirus infections.
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影响因子：
6.7
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影响因子：
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