Noordhoek Gerda T, Weel Jan F L, Poelstra Elisabeth, Hooghiemstra Marijke, Brandenburg Afke H
Public Health Laboratory Friesland, Post Box 21020, 8900 JA Leeuwarden, The Netherlands.
J Clin Virol. 2008 Feb;41(2):75-80. doi: 10.1016/j.jcv.2007.09.011.
Enteroviruses (EV) and parechoviruses (HPeV) are the most common causes of aseptic meningitis, encephalitis and sepsis-like syndrome in neonates. Detection by nucleic acid amplification methods improves patient management.
Development of a real-time PCR assay on a LightCycler for simultaneous detection of EV, HPeV and an internal control to monitor inhibition.
We investigated the value of the new assay, prospectively, in a variety of samples from patients suspected of having viral meningitis or sepsis-like syndrome.
The assay detected 64 EV serotypes and HPeV types 1-4. Of 186 patients, 63 (33.9%) were EV positive and 18 (9.7%) HPeV positive in one or more samples. In 43 of 159 feces and 6 of 57 throat samples viral culture and PCR were positive. With real-time PCR 27 extra EV and 19 HPeV positives were found. Blood and CSF were present from 33 patients. In 19 patients blood and CSF were positive, one was only positive in CSF, two were only positive in blood, 11 were negative. From 96 patients CSF and/or blood samples were tested and compared to results in throat and/or feces samples. Forty patients were EV-PCR and 14 HPeV-PCR positive in blood and/or CSF. All of these were confirmed by a positive PCR for the respective virus in feces and/or throat.
Simultaneous detection of EV and HPeV with this two-step real-time PCR is specific, faster and more sensitive than viral culture. All systemic infections (blood or CSF positive) were confirmed in feces. Culture is no longer necessary for clinical diagnosis and should only be performed on PCR-positive samples to obtain isolates for typing purposes. Application of this assay is an important improvement for patient management since the outcome of the analysis is available within the time frame of clinical decision-making.
肠道病毒（EV）和帕利病毒（HPeV）是新生儿无菌性脑膜炎、脑炎和败血症样综合征最常见的病因。通过核酸扩增方法进行检测可改善患者管理。
开发一种在LightCycler上的实时PCR检测方法，用于同时检测EV、HPeV和一种内部对照以监测抑制情况。
我们前瞻性地研究了这种新检测方法在各种疑似患有病毒性脑膜炎或败血症样综合征患者样本中的价值。
该检测方法可检测64种EV血清型和1 - 4型HPeV。在186例患者中，63例（33.9%）在一个或多个样本中EV呈阳性，18例（9.7%）HPeV呈阳性。在159份粪便样本中的43份以及57份咽喉样本中的6份，病毒培养和PCR检测均呈阳性。通过实时PCR又发现了27例额外的EV阳性和19例HPeV阳性。33例患者提供了血液和脑脊液样本。19例患者的血液和脑脊液呈阳性，1例仅脑脊液呈阳性，2例仅血液呈阳性，11例呈阴性。对96例患者的脑脊液和/或血液样本进行检测，并与咽喉和/或粪便样本的检测结果进行比较。40例患者的血液和/或脑脊液中EV-PCR呈阳性，14例HPeV-PCR呈阳性。所有这些均通过粪便和/或咽喉中相应病毒的PCR阳性得到证实。
这种两步实时PCR同时检测EV和HPeV具有特异性，比病毒培养更快、更灵敏。所有全身性感染（血液或脑脊液呈阳性）在粪便中均得到证实。临床诊断不再需要进行病毒培养，仅应对PCR阳性样本进行培养以获取用于分型的分离株。由于分析结果可在临床决策的时间范围内获得，因此该检测方法的应用对患者管理是一项重要的改进。
