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从临床粪便样本中检测人类肠道病毒和人类副肠道病毒(HPeV)基因型:聚合酶链反应和直接分子分型、培养特性和血清分型。

Detection of human enterovirus and human parechovirus (HPeV) genotypes from clinical stool samples: polymerase chain reaction and direct molecular typing, culture characteristics, and serotyping.

机构信息

Department of Medical Microbiology, Laboratory of Clinical Virology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.

出版信息

Diagn Microbiol Infect Dis. 2010 Oct;68(2):166-73. doi: 10.1016/j.diagmicrobio.2010.05.016.

Abstract

Molecular (polymerase chain reaction [PCR]) methods are increasingly used to detect and type human enteroviruses (HEVs) and parechoviruses (HPeV). Here, we assessed their value in comparison to virus culture and serotyping for detection and typing of HEV and HPeV in stool samples from hospitalized patients. By use of real-time PCR, 221/1174 patients (18.8%) were found positive for HEV/HPeV. By cell culture, a virus could be isolated from 107 of the HEV/HPeV PCR-positive samples. Culture efficiency was correlated to the Ct value, (geno)type, and cell lines used. Of the HEV/HPeV PCR-positive samples, 47% could be genotyped by VP1 genotyping and 25% by serotyping. In conclusion, PCR detection of HEV/HPeV from stool is more sensitive than virus culture, particularly for coxsackieviruses A and HPeVs. However, the genotyping method used here could identify only 47% of the HEV/HPeV strains. Further optimization and validation of direct genotyping are needed, and clinical relevance of HEV/HPeV detection in stool needs to be determined.

摘要

分子(聚合酶链反应 [PCR])方法越来越多地用于检测和分型人类肠道病毒(HEV)和肠道病毒 71 型(HPeV)。在这里,我们评估了它们在检测和分型粪便样本中的 HEV 和 HPeV 方面与病毒培养和血清分型相比的价值。通过实时 PCR,在 1174 名住院患者中有 221 名(18.8%)为 HEV/HPeV 阳性。通过细胞培养,可以从 107 份 HEV/HPeV PCR 阳性样本中分离出病毒。培养效率与 Ct 值、(基因)型和使用的细胞系相关。在 HEV/HPeV PCR 阳性样本中,47%可以通过 VP1 基因分型,25%可以通过血清分型。总之,粪便中 HEV/HPeV 的 PCR 检测比病毒培养更敏感,特别是对柯萨奇病毒 A 和 HPeVs。然而,这里使用的基因分型方法只能识别 47%的 HEV/HPeV 株。需要进一步优化和验证直接基因分型,并且需要确定粪便中 HEV/HPeV 检测的临床意义。

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