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Development of Prototype Filovirus Recombinant Antigen Immunoassays.


ABSTRACT:

Background

Throughout the 2014-2015 Ebola outbreak in West Africa, major gaps were exposed in the availability of validated rapid diagnostic platforms, protective vaccines, and effective therapeutic agents. These gaps potentiated the development of prototype rapid lateral flow immunodiagnostic (LFI) assays that are true point-of-contact platforms, for the detection of active Ebola infections in small blood samples.

Methods

Recombinant Ebola and Marburg virus matrix VP40 and glycoprotein (GP) antigens were used to derive a panel of monoclonal and polyclonal antibodies. Antibodies were tested using a multivariate approach to identify antibody-antigen combinations suitable for enzyme-linked immunosorbent assay (ELISA) and LFI assay development.

Results

Polyclonal antibodies generated in goats were superior reagents for capture and detection of recombinant VP40 in test sample matrices. These antibodies were optimized for use in antigen-capture ELISA and LFI assay platforms. Prototype immunoglobulin M (IgM)/immunoglobulin G (IgG) ELISAs were similarly developed that specifically detect Ebola virus-specific antibodies in the serum of experimentally infected nonhuman primates and in blood samples obtained from patients with Ebola from Sierra Leone.

Conclusions

The prototype recombinant Ebola LFI assays developed in these studies have sensitivities that are useful for clinical diagnosis of acute ebolavirus infections. The antigen-capture and IgM/IgG ELISAs provide additional confirmatory assay platforms for detecting VP40 and other ebolavirus-specific immunoglobulins.

SUBMITTER: Boisen ML 

PROVIDER: S-EPMC4564556 | biostudies-literature | 2015 Oct

REPOSITORIES: biostudies-literature

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Development of Prototype Filovirus Recombinant Antigen Immunoassays.

Boisen Matt L ML   Oottamasathien Darin D   Jones Abigail B AB   Millett Molly M MM   Nelson Diana S DS   Bornholdt Zachary A ZA   Fusco Marnie L ML   Abelson Dafna M DM   Oda Shun-Ichiro S   Hartnett Jessica N JN   Rowland Megan M MM   Heinrich Megan L ML   Akdag Marjan M   Goba Augustine A   Momoh Mambu M   Fullah Mohammed M   Baimba Francis F   Gbakie Michael M   Safa Sadiki S   Fonnie Richard R   Kanneh Lansana L   Cross Robert W RW   Geisbert Joan B JB   Geisbert Thomas W TW   Kulakosky Peter C PC   Grant Donald S DS   Shaffer Jeffery G JG   Schieffelin John S JS   Wilson Russell B RB   Saphire Erica Ollmann EO   Branco Luis M LM   Garry Robert F RF   Khan S Humarr SH   Pitts Kelly R KR  

The Journal of infectious diseases 20150730


<h4>Background</h4>Throughout the 2014-2015 Ebola outbreak in West Africa, major gaps were exposed in the availability of validated rapid diagnostic platforms, protective vaccines, and effective therapeutic agents. These gaps potentiated the development of prototype rapid lateral flow immunodiagnostic (LFI) assays that are true point-of-contact platforms, for the detection of active Ebola infections in small blood samples.<h4>Methods</h4>Recombinant Ebola and Marburg virus matrix VP40 and glycop  ...[more]

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