Detection of all known parechoviruses by real-time PCR
Detection of all known parechoviruses by real-time PCR
复制标题
DOI：
10.1128/jcm.00277-08
复制
发表时间：
2008-08-01
影响因子：
9.4
通讯作者：
Oberste, M. Steven
中科院分区：
文献类型：
--
作者：
Nix, W. Allan;Maher, Kaija;Oberste, M. Steven
小核糖核酸病毒科的双埃柯病毒属包含两个物种，即人类双埃柯病毒（HPeV）和Ljungan病毒（LV）。HPeV（包括以前的埃可病毒22和23，现在分别是HPeV 1型（HPeV 1）和HPeV 2）引起广泛的疾病，包括无菌性脑膜炎，胃肠炎，脑炎，急性呼吸道疾病和新生儿败血症样疾病。在寻找与人类致命性心肌炎病例相关的传染性病原体期间，从瑞典的银行田鼠中分离出了LV。由于使用细胞培养和中和来研究肠道病毒样疾病的减少，目前很少有实验室有能力检测双埃柯病毒。我们已经开发了一种实时逆转录-PCR（RT-PCR）检测所有已知的成员属副肠孤病毒。该检测试剂靶向双埃柯病毒基因组的5'非翻译区（5' NTR）中的保守区，可同时检测HPeV和LV，而其他已发表的双埃柯病毒5 'NTR检测试剂仅检测已知的HPeV或仅检测LV。当需要时，HPeV和LV可以通过对5 'NTR实时RT-PCR扩增子进行测序来区分。该检测试剂盒的灵敏度约为细胞培养的100倍，可用于检测原始临床标本。广泛特异性PCR方法的可用性应有助于检测新的人类双埃柯病毒，以及其他哺乳动物物种中的新的双埃柯病毒，并提供了一个机会，调查这些病毒在人类和动物疾病中的作用。
The Parechovirus genus of the Picornaviridae family contains two species, Human parechovirus (HPeV) and Ljungan virus (LV). The HPeVs (including the former echoviruses 22 and 23, now HPeV type 1 (HPeV1) and HPeV2, respectively) cause a wide spectrum of disease, including aseptic meningitis, gastroenteritis, encephalitis, acute respiratory illness, and neonatal sepsis-like disease. The LVs were isolated from bank voles in Sweden during a search for an infectious agent linked to fatal myocarditis cases in humans. Because of the decline in use of cell culture and neutralization to investigate enterovirus-like disease, very few laboratories currently have the capability to test for parechoviruses. We have developed a real-time reverse transcription-PCR (RT-PCR) assay for detection of all known members of the genus Parechovirus. The assay targets the conserved regions in the 5' nontranslated region (5'NTR) of the parechovirus genome and can detect both HPeVs and LVs, unlike other published parechovirus 5'NTR assays, which only detect known HPeVs or only LVs. HPeV and LV can be differentiated by sequencing the 5'NTR real-time RT-PCR amplicon, when needed. The assay is approximately 100 times more sensitive than cell culture and may be used to test original clinical specimens. The availability of a broad-specificity PCR method should facilitate the detection of new human parechoviruses, as well as new parechoviruses in other mammalian species, and provide an opportunity to investigate the role of these viruses in human and animal disease.