title: Development of a Sensitive and Specific Serological Assay Based on Luminex Technology for Detection of Antibodies to Zaire Ebola Virus.
abstract: 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.
authors: IRD UMI 233-INSERM U1175-Montpellier University, Montpellier, France Iahidjo.ayouba@ird.fr.
authors: Ahidjo
authors: 0000-0002-5081-1632
authors: A
authors: Ayouba
authors: Chaire de santé publique, Département de Pharmacie, Université de Conakry, Conakry, Guinea.
authors: Abdoulaye
authors: A
authors: Touré
authors: IRD UMI 233-INSERM U1175-Montpellier University, Montpellier, France.
authors: Christelle
authors: C
authors: Butel
authors: IRD UMI 233-INSERM U1175-Montpellier University, Montpellier, France.
authors: Alpha Kabinet
authors: AK
authors: Keita
authors: IRD UMI 233-INSERM U1175-Montpellier University, Montpellier, France.
authors: Florian
authors: F
authors: Binetruy
authors: Donka National Hospital, Conakry, Guinea.
authors: Mamadou S
authors: MS
authors: Sow
authors: INSERM U1058-Montpellier University, Montpellier, France.
authors: Vincent
authors: V
authors: Foulongne
authors: IRD UMI 233-INSERM U1175-Montpellier University, Montpellier, France.
authors: Eric
authors: E
authors: Delaporte
authors: IRD UMI 233-INSERM U1175-Montpellier University, Montpellier, France.
authors: Martine
authors: M
authors: Peeters
mesh_terms: D000349:Africa; D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000276:immunology*; D003429:Cross Reactions; D029043:Ebolavirus / Q000276:immunology*; D005602:France; D019142:Hemorrhagic Fever, Ebola / Q000175:diagnosis*; D006801:Humans; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral
keywords: Ebola virus; Luminex; recombinant proteins; serology