Ayouba Ahidjo, Touré Abdoulaye, Butel Christelle, Keita Alpha Kabinet, Binetruy Florian, Sow Mamadou S, Foulongne Vincent, Delaporte Eric, Peeters Martine
IRD UMI 233-INSERM U1175-Montpellier University, Montpellier, France
Chaire de santé publique, Département de Pharmacie, Université de Conakry, Conakry, Guinea.
J Clin Microbiol. 2016 Dec 28;55(1):165-176. doi: 10.1128/JCM.01979-16. Print 2017 Jan.
The recent Zaire Ebola virus (EBOV) outbreak in West Africa illustrates clearly the need for additional studies with humans and animals to elucidate the ecology of Ebola viruses (EBVs). In this study, we developed a serological assay based on the Luminex technology. Nine recombinant proteins representing different viral regions (nucleoprotein [NP], 40-kDa viral protein [VP40], and glycoprotein [GP]) from four of the five EBV lineages were used. Samples from 94 survivors of the EBOV outbreak in Guinea and negative samples from 108 patients in France were used to calculate test performance for EBOV detection and cross-reaction with other Ebola virus lineages. For EBOV antibody detection, sensitivities of 95.7%, 96.8%, and 92.5% and specificities of 94.4%, 95.4%, and 96.3% for NP, GP, and VP40, respectively, were observed. All EBOV-negative samples that presented a reaction, except for one, interacted with a single antigen, whereas almost all samples from EBOV survivors were simultaneously reactive with NP and GP (90/94) or with NP, GP, and VP40 (87/94). Considering as positive for past EBOV infection only samples that reacted with EBOV NP and GP, sensitivity was 95.7% and specificity increased to 99.1%. Comparing results with commercial EBOV NP and GP enzyme-linked immunosorbent assays (ELISAs; Alpha Diagnostic, San Antonio, TX), lower sensitivity (92.5%) and high specificity (100%) were observed with the same positivity criteria. Samples from EBOV survivors cross-reacted with GP from Sudan Ebola virus (GP-SUDV) (81.9%), GP from Bundibugyo Ebola virus (GP-BDBV) (51.1%), GP from Reston Ebola virus (GP-RESTV) (9.6%), VP40-SUDV (76.6%), and VP40-BDBV (38.3%). Overall, we developed a sensitive and specific high-throughput serological assay, and defined an algorithm, for epidemiological surveys with humans.
近期在西非爆发的扎伊尔埃博拉病毒（EBOV）疫情清楚地表明，有必要对人类和动物开展更多研究，以阐明埃博拉病毒（EBV）的生态学。在本研究中，我们基于Luminex技术开发了一种血清学检测方法。使用了代表五种EBV谱系中四种不同病毒区域（核蛋白[NP]、40 kDa病毒蛋白[VP40]和糖蛋白[GP]）的九种重组蛋白。来自几内亚EBOV疫情94名幸存者的样本以及来自法国108名患者的阴性样本用于计算EBOV检测的检测性能以及与其他埃博拉病毒谱系的交叉反应。对于EBOV抗体检测，观察到NP、GP和VP40的敏感性分别为95.7%、96.8%和92.5%，特异性分别为94.4%、95.4%和96.3%。除一个样本外，所有出现反应的EBOV阴性样本均与单一抗原相互作用，而几乎所有来自EBOV幸存者的样本同时与NP和GP（90/94）或与NP、GP和VP40（87/94）反应。仅将与EBOV NP和GP反应的样本视为既往EBOV感染阳性，敏感性为95.7%，特异性提高到99.1%。将结果与商用EBOV NP和GP酶联免疫吸附测定（ELISA；Alpha Diagnostic，圣安东尼奥，德克萨斯州）进行比较，在相同阳性标准下观察到较低的敏感性（92.5%）和较高的特异性（100%）。来自EBOV幸存者的样本与苏丹埃博拉病毒的GP（GP-SUDV）（81.9%）、本迪布焦埃博拉病毒的GP（GP-BDBV）（51.1%）、莱斯顿埃博拉病毒的GP（GP-RESTV）（9.6%）、VP40-SUDV（76.6%）和VP40-BDBV（38.3%）发生交叉反应。总体而言，我们开发了一种灵敏且特异的高通量血清学检测方法，并定义了一种算法，用于人类流行病学调查。
