Development of Prototype Filovirus Recombinant Antigen Immunoassays

Boisen, Matthew L.; Oottamasathien, Darin; Jones, Abigail B.; Millett, Molly M.; Nelson, Diana; Bornholdt, Zachary A.; Fusco, Marnie L.; Abelson, Dafna M.; Oda, Shun-ichiro; Hartnett, Jessica N.; Rowland, Megan M.; Heinrich, Megan L.; Akdag, Marjan; Goba, Augustine; Momoh, Mambu; Fullah, Mohammed; Baimba, Francis; Gbakie, Michael; Safa, Sadiki; Fonnie, Richard; Kanneh, Lansana; Cross, Robert W.; Geisbert, Joan B.; Geisbert, Thomas W.; Kulakosky, Peter C.; Grant, Donald S.; Shaffer, Jeffery G.; Schieffelin, John S.; Wilson, Russell B.; Saphire, Erica Ollmann; Branco, Luis M.; Garry, Robert F.; Khan, Sheik Humarr; Pitts, Kelly R.

doi:10.1093/infdis/jiv353

Cited by 31 publications

(19 citation statements)

References 26 publications

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“…LCMS technology is also not feasible in field clinics in West Africa where Lassa fever patients are prevalent. Therefore, simple assays such as dipstick style chromatographic assays, including lateral flow immunoassays, have gained acceptance, and can be conducted with minimal resources and training [ 35 , 58 – 60 ]. Several small molecules with high biomarker potential were identified including adducts of the modified nucleoside 1-methylinosine.…”

Section: Discussionmentioning

confidence: 99%

Metabolomics analyses identify platelet activating factors and heme breakdown products as Lassa fever biomarkers

et al. 2017

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Lassa fever afflicts tens of thousands of people in West Africa annually. The rapid progression of patients from febrile illness to fulminant syndrome and death provides incentive for development of clinical prognostic markers that can guide case management. The small molecule profile of serum from febrile patients triaged to the Viral Hemorrhagic Fever Ward at Kenema Government Hospital in Sierra Leone was assessed using untargeted Ultra High Performance Liquid Chromatography Mass Spectrometry. Physiological dysregulation resulting from Lassa virus (LASV) infection occurs at the small molecule level. Effects of LASV infection on pathways mediating blood coagulation, and lipid, amino acid, nucleic acid metabolism are manifest in changes in the levels of numerous metabolites in the circulation. Several compounds, including platelet activating factor (PAF), PAF-like molecules and products of heme breakdown emerged as candidates that may prove useful in diagnostic assays to inform better care of Lassa fever patients.

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Section: Discussionmentioning

confidence: 99%

Metabolomics analyses identify platelet activating factors and heme breakdown products as Lassa fever biomarkers

et al. 2017

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“…LASV RDTs, which leverage the same antibody/antigen capture agents as an ELISA but packaged in a stripped-down lateral flow format, can play an important role for patient care and outbreak response in outlying laboratories and clinics. A dipstick-based RDT for LASV has been developed that detects NP from fingerstick whole blood specimens 7 108 134 135. For LASV lineage IV (Josiah strain), the dipstick LASV RDT performed with good sensitivity (91% sensitivity, 86% specificity) compared with its progenitor ELISA (94% sensitivity, 84% specificity; both relative to qPCR).…”

Section: Lassa Diagnosticsmentioning

confidence: 99%

Diagnostics for Lassa fever virus: a genetically diverse pathogen found in low-resource settings

Mazzola

Kelly-Cirino

2019

BMJ Glob Health

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Lassa fever virus (LASV) causes acute viral haemorrhagic fever with symptoms similar to those seen with Ebola virus infections. LASV is endemic to West Africa and is transmitted through contact with excretions of infected Mastomys natalensis rodents and other rodent species. Due to a high fatality rate, lack of treatment options and difficulties with prevention and control, LASV is one of the high-priority pathogens included in the WHO R&D Blueprint. The WHO LASV vaccine strategy relies on availability of effective diagnostic tests. Current diagnostics for LASV include in-house and commercial (primarily research-only) laboratory-based serological and nucleic acid amplification tests. There are two commercially available (for research use only) rapid diagnostic tests (RDTs), and a number of multiplex panels for differential detection of LASV infection from other endemic diseases with similar symptoms have been evaluated. However, a number of diagnostic gaps remain. Lineage detection is a challenge due to the genomic diversity of LASV, as pan-lineage sensitivity for both molecular and immunological detection is necessary for surveillance and outbreak response. While pan-lineage ELISA and RDTs are commercially available (for research use only), validation and external quality assessment (EQA) is needed to confirm detection sensitivity for all known or relevant strains. Variable sensitivity of LASV PCR tests also highlights the need for improved validation and EQA. Given that LASV outbreaks typically occur in low-resource settings, more options for point-of-care testing would be valuable. These requirements should be taken into account in target product profiles for improved LASV diagnostics.

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“…Prior to the 2014 emergence of EVD in Guinea, ReEBOV RDT prototypes were developed using rVP40 and rNP antigens and respective polyclonal and monoclonal antibodies [16]. In June 2014, field testing of rVP40 and rNP enzyme-linked immunosorbent assays demonstrated strong reactivity of serum from patients with suspected EVD to these recombinant protein and antibody reagents, further strengthening confidence in the necessary cross-reactivity of the antibody reagents with the circulating strain of EBOV [17]. Ultimately, the decision was made to expedite RDT development toward detection of EBOV VP40 antigen, owing to reagent availability and promising feasibility studies.…”

Section: Ebola Critical Reagent Developmentmentioning

confidence: 99%

Analytical Validation of the ReEBOV Antigen Rapid Test for Point-of-Care Diagnosis of Ebola Virus Infection

et al. 2016

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Background. Ebola virus disease (EVD) is a severe viral illness caused by Ebola virus (EBOV). The 2013-2016 EVD outbreak inWest Africa is the largest recorded, with >11 000 deaths. Development of the ReEBOV Antigen Rapid Test (ReEBOV RDT) was expedited to provide a point-of-care test for suspected EVD cases.Methods. Recombinant EBOV viral protein 40 antigen was used to derive polyclonal antibodies for RDT and enzyme-linked immunosorbent assay development. ReEBOV RDT limits of detection (LOD), specificity, and interference were analytically validated on the basis of Food and Drug Administration (FDA) guidance.Results. The ReEBOV RDT specificity estimate was 95% for donor serum panels and 97% for donor whole-blood specimens. The RDT demonstrated sensitivity to 3 species of Ebolavirus (Zaire ebolavirus, Sudan ebolavirus, and Bundibugyo ebolavirus) associated with human disease, with no cross-reactivity by pathogens associated with non-EBOV febrile illness, including malaria parasites. Interference testing exhibited no reactivity by medications in common use. The LOD for antigen was 4.7 ng/test in serum and 9.4 ng/test in whole blood. Quantitative reverse transcription-polymerase chain reaction testing of nonhuman primate samples determined the range to be equivalent to 3.0 × 10 5 -9.0 × 10 8 genomes/mL.Conclusions. The analytical validation presented here contributed to the ReEBOV RDT being the first antigen-based assay to receive FDA and World Health Organization emergency use authorization for this EVD outbreak, in February 2015.

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Development of Prototype Filovirus Recombinant Antigen Immunoassays

Cited by 31 publications

References 26 publications

Metabolomics analyses identify platelet activating factors and heme breakdown products as Lassa fever biomarkers

Metabolomics analyses identify platelet activating factors and heme breakdown products as Lassa fever biomarkers

Diagnostics for Lassa fever virus: a genetically diverse pathogen found in low-resource settings

Analytical Validation of the ReEBOV Antigen Rapid Test for Point-of-Care Diagnosis of Ebola Virus Infection

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