Standardization of Hepatitis E Virus (HEV) Nucleic Acid Amplification Technique-Based Assays: an Initial Study To Evaluate a Panel of HEV Strains and Investigate Laboratory Performance
Standardization of Hepatitis E Virus (HEV) Nucleic Acid Amplification Technique-Based Assays: an Initial Study To Evaluate a Panel of HEV Strains and Investigate Laboratory Performance
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DOI：
10.1128/jcm.02578-10
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发表时间：
2011-04-01
影响因子：
9.4
通讯作者：
Nuebling, C. Micha
中科院分区：
文献类型：
--
作者：
Baylis, Sally A.;Hanschmann, Kay-Martin;Nuebling, C. Micha
利用一组含戊型肝炎病毒的血浆样本，研究了戊型肝炎病毒（HEV） RNA核酸扩增（NAT）检测方法的性能。该小组包括22份HEV阳性血浆样本，代表从献血者获得的HEV基因型3a、3b、3f和4c的10倍系列稀释。包括两份阴性对照血浆样本。所有样本均采用盲法。血浆样品被制成液体/冷冻材料，并在干冰上分发给参与者。要求实验室使用其常规的HEV检测方法对该小组进行检测，并对样品进行阳性或阴性评分，并可选择返回以拷贝数/ml为单位的HEV RNA数据。来自10个不同国家的20个实验室参与了这项研究。数据由所有参与实验室返回；10个实验室返回了定量数据。除一项检测外，所有检测均采用常规或实时逆转录酶PCR （RT-PCR）方法进行内部开发。与病毒株无关，大多数检测方法的灵敏度相差100到1000倍。虽然定量数据有限，但在相似于6 ~ 4 log(10) copies/ml范围内的样品，其几何均值的标准差在0.38 ~ 1.09之间。除了一个模棱两可的结果外，阴性样品中未检测到HEV RNA。检测灵敏度的可变性突出了HEV RNA NAT检测标准化的必要性。
The performance of hepatitis E virus (HEV) RNA nucleic acid amplification (NAT)-based assays has been investigated using a panel of HEV-containing plasma samples. The panel comprised 22 HEV-positive plasma samples representing 10-fold serial dilutions of HEV genotypes 3a, 3b, 3f, and 4c obtained from blood donors. Two negative-control plasma samples were included. All samples were blinded. The plasma samples were prepared as liquid/frozen materials and distributed to participants on dry ice. Laboratories were requested to test the panel using their routine HEV assays and to score samples as either positive or negative and could optionally return data in copies/ml for HEV RNA. Twenty laboratories from 10 different countries participated in the study. Data were returned by all participating laboratories; 10 laboratories returned quantitative data. All assays except one were developed in-house using conventional or real-time reverse transcriptase PCR (RT-PCR) methodologies. There was a 100- to 1,000-fold difference in sensitivity between the majority of assays, independent of the virus strain. Although the quantitative data were limited, for the samples in the range of similar to 6 to 4 log(10) copies/ml, the standard deviations of the geometric means of the samples ranged between 0.38 and 1.09. Except for one equivocal result, HEV RNA was not detected in the negative samples. The variability of assay sensitivity highlights the need for the standardization of HEV RNA NAT assays.