Rapid diagnosis of Ebola hemorrhagic fever by reverse transcription-PCR in an outbreak setting and assessment of patient viral load as a predictor of outcome
Rapid diagnosis of Ebola hemorrhagic fever by reverse transcription-PCR in an outbreak setting and assessment of patient viral load as a predictor of outcome
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DOI：
10.1128/jvi.78.8.4330-4341.2004
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发表时间：
2004-04-01
影响因子：
5.4
通讯作者：
Nichol, ST
中科院分区：
文献类型：
--
作者：
Towner, JS;Rollin, PE;Nichol, ST
有记录以来最大规模的埃博拉出血热（EHF）疫情于2000年8月至2001年1月在乌干达爆发。疫情集中在乌干达北部的古卢地区，并继发传播到其他地区。南非约翰内斯堡国家病毒学研究所初步诊断出苏丹埃博拉病毒后，在圣玛丽拉科尔医院古卢区建立了一个临时诊断实验室。该实验室使用抗原捕获和逆转录聚合酶链反应（RT-PCR）来诊断可疑患者的苏丹埃博拉病毒感染。事实证明，RT-PCR 和抗原捕获诊断检测对于检测患者血清、血浆和全血中的埃博拉病毒非常有效。在感染过程早期收集的样本中，RT-PCR 检测可以在抗原捕获检测前 24 至 48 小时检测到埃博拉病毒。收集了 1000 多份血液样本，在整个感染过程中从许多患者身上获取了多个样本。使用实时定量 RT-PCR 测定致命和非致命病例患者的多个样本中的病毒载量，这些数据与疾病结果相关。死亡患者的 RNA 拷贝水平平均比存活患者高 2 log(10)。使用来自多名 EHF 患者的临床材料，我们对糖蛋白的可变区进行了测序。这种苏丹埃博拉病毒毒株并非源自早期的 Boniface (1976) 或 Maleo (1979) 毒株，但它与两者具有共同的祖先。此外，序列和流行病学数据都与该疫情源自单次引入人群一致。
The largest outbreak on record of Ebola hemorrhagic fever (EHF) occurred in Uganda from August 2000 to January 2001. The outbreak was centered in the Gulu district of northern Uganda, with secondary transmission to other districts. After the initial diagnosis of Sudan ebolavirus by the National Institute for Virology in Johannesburg, South Africa, a temporary diagnostic laboratory was established within the Gulu district at St. Mary's Lacor Hospital. The laboratory used antigen capture and reverse transcription-PCR (RT-PCR) to diagnose Sudan ebolavirus infection in suspect patients. The RT-PCR and antigen-capture diagnostic assays proved very effective for detecting ebolavirus in patient serum, plasma, and whole blood. In samples collected very early in the course of infection, the RT-PCR assay could detect ebolavirus 24 to 48 h prior to detection by antigen capture. More than 1,000 blood samples were collected, with multiple samples obtained from many patients throughout the course of infection. Real-time quantitative RT-PCR was used to determine the viral load in multiple samples from patients with fatal and nonfatal cases, and these data were correlated with the disease outcome. RNA copy levels in patients who died averaged 2 log(10) higher than those in patients who survived. Using clinical material from multiple EHF patients, we sequenced the variable region of the glycoprotein. This Sudan ebolavirus strain was not derived from either the earlier Boniface (1976) or Maleo (1979) strain, but it shares a common ancestor with both. Furthermore, both sequence and epidemiologic data are consistent with the outbreak having originated from a single introduction into the human population.