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

Robert Cross, Matthew L. Boisen, Molly M. Millett, DIana S. Nelson, Darin Oottamasathien, Jessica N. Hartnett, Abigal B. Jones, Augustine Goba, Mambu Momoh, Mohamed Fullah, Zachary A. Bornholdt, Marnie L. Fusco, Dafna M. Abelson, Shunichiro Oda, Bethany L. Brown, Ha Pham, Megan M. Rowland, Krystle N. Agans, Joan B. Geisbert, Megan L. Heinrich & 14 others Peter C. Kulakosky, Jeffrey G. Shaffer, John S. Schieffelin, Brima Kargbo, Momoh Gbetuwa, Sahr M. Gevao, Russell B. Wilson, Erica Ollmann Saphire, Kelly R. Pitts, Sheik Humarr Khan, Donald S. Grant, Thomas Geisbert, Luis M. Branco, Robert F. Garry

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Background. Ebola virus disease (EVD) is a severe viral illness caused by Ebola virus (EBOV). The 2013-2016 EVD outbreak in West 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 × 105-9.0 × 108 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.

Original languageEnglish (US)
Pages (from-to)S210-S217
JournalJournal of Infectious Diseases
Volume214
DOIs
StatePublished - Oct 15 2016

Fingerprint

Ebola Hemorrhagic Fever
Point-of-Care Systems
Ebolavirus
Antigens
United States Food and Drug Administration
Disease Outbreaks
Limit of Detection
Democratic Republic of the Congo
Sudan
Western Africa
Viral Proteins
Blood Donors
Serum
Primates
Malaria
Reverse Transcription
Parasites
Emergencies
Fever
Enzyme-Linked Immunosorbent Assay

Keywords

  • Diagnostic
  • Ebola, point of care
  • Lateral flow immunoassay

ASJC Scopus subject areas

  • Immunology and Allergy
  • Infectious Diseases

Cite this

Cross, R., Boisen, M. L., Millett, M. M., Nelson, DI. S., Oottamasathien, D., Hartnett, J. N., ... Garry, R. F. (2016). Analytical Validation of the ReEBOV Antigen Rapid Test for Point-of-Care Diagnosis of Ebola Virus Infection. Journal of Infectious Diseases, 214, S210-S217. https://doi.org/10.1093/infdis/jiw293

Analytical Validation of the ReEBOV Antigen Rapid Test for Point-of-Care Diagnosis of Ebola Virus Infection. / Cross, Robert; Boisen, Matthew L.; Millett, Molly M.; Nelson, DIana S.; Oottamasathien, Darin; Hartnett, Jessica N.; Jones, Abigal B.; Goba, Augustine; Momoh, Mambu; Fullah, Mohamed; Bornholdt, Zachary A.; Fusco, Marnie L.; Abelson, Dafna M.; Oda, Shunichiro; Brown, Bethany L.; Pham, Ha; Rowland, Megan M.; Agans, Krystle N.; Geisbert, Joan B.; Heinrich, Megan L.; Kulakosky, Peter C.; Shaffer, Jeffrey G.; Schieffelin, John S.; Kargbo, Brima; Gbetuwa, Momoh; Gevao, Sahr M.; Wilson, Russell B.; Saphire, Erica Ollmann; Pitts, Kelly R.; Khan, Sheik Humarr; Grant, Donald S.; Geisbert, Thomas; Branco, Luis M.; Garry, Robert F.

In: Journal of Infectious Diseases, Vol. 214, 15.10.2016, p. S210-S217.

Research output: Contribution to journalArticle

Cross, R, Boisen, ML, Millett, MM, Nelson, DIS, Oottamasathien, D, Hartnett, JN, Jones, AB, Goba, A, Momoh, M, Fullah, M, Bornholdt, ZA, Fusco, ML, Abelson, DM, Oda, S, Brown, BL, Pham, H, Rowland, MM, Agans, KN, Geisbert, JB, Heinrich, ML, Kulakosky, PC, Shaffer, JG, Schieffelin, JS, Kargbo, B, Gbetuwa, M, Gevao, SM, Wilson, RB, Saphire, EO, Pitts, KR, Khan, SH, Grant, DS, Geisbert, T, Branco, LM & Garry, RF 2016, 'Analytical Validation of the ReEBOV Antigen Rapid Test for Point-of-Care Diagnosis of Ebola Virus Infection', Journal of Infectious Diseases, vol. 214, pp. S210-S217. https://doi.org/10.1093/infdis/jiw293
Cross, Robert ; Boisen, Matthew L. ; Millett, Molly M. ; Nelson, DIana S. ; Oottamasathien, Darin ; Hartnett, Jessica N. ; Jones, Abigal B. ; Goba, Augustine ; Momoh, Mambu ; Fullah, Mohamed ; Bornholdt, Zachary A. ; Fusco, Marnie L. ; Abelson, Dafna M. ; Oda, Shunichiro ; Brown, Bethany L. ; Pham, Ha ; Rowland, Megan M. ; Agans, Krystle N. ; Geisbert, Joan B. ; Heinrich, Megan L. ; Kulakosky, Peter C. ; Shaffer, Jeffrey G. ; Schieffelin, John S. ; Kargbo, Brima ; Gbetuwa, Momoh ; Gevao, Sahr M. ; Wilson, Russell B. ; Saphire, Erica Ollmann ; Pitts, Kelly R. ; Khan, Sheik Humarr ; Grant, Donald S. ; Geisbert, Thomas ; Branco, Luis M. ; Garry, Robert F. / Analytical Validation of the ReEBOV Antigen Rapid Test for Point-of-Care Diagnosis of Ebola Virus Infection. In: Journal of Infectious Diseases. 2016 ; Vol. 214. pp. S210-S217.
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abstract = "Background. Ebola virus disease (EVD) is a severe viral illness caused by Ebola virus (EBOV). The 2013-2016 EVD outbreak in West 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 × 105-9.0 × 108 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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author = "Robert Cross and Boisen, {Matthew L.} and Millett, {Molly M.} and Nelson, {DIana S.} and Darin Oottamasathien and Hartnett, {Jessica N.} and Jones, {Abigal B.} and Augustine Goba and Mambu Momoh and Mohamed Fullah and Bornholdt, {Zachary A.} and Fusco, {Marnie L.} and Abelson, {Dafna M.} and Shunichiro Oda and Brown, {Bethany L.} and Ha Pham and Rowland, {Megan M.} and Agans, {Krystle N.} and Geisbert, {Joan B.} and Heinrich, {Megan L.} and Kulakosky, {Peter C.} and Shaffer, {Jeffrey G.} and Schieffelin, {John S.} and Brima Kargbo and Momoh Gbetuwa and Gevao, {Sahr M.} and Wilson, {Russell B.} and Saphire, {Erica Ollmann} and Pitts, {Kelly R.} and Khan, {Sheik Humarr} and Grant, {Donald S.} and Thomas Geisbert and Branco, {Luis M.} and Garry, {Robert F.}",
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T1 - Analytical Validation of the ReEBOV Antigen Rapid Test for Point-of-Care Diagnosis of Ebola Virus Infection

AU - Cross, Robert

AU - Boisen, Matthew L.

AU - Millett, Molly M.

AU - Nelson, DIana S.

AU - Oottamasathien, Darin

AU - Hartnett, Jessica N.

AU - Jones, Abigal B.

AU - Goba, Augustine

AU - Momoh, Mambu

AU - Fullah, Mohamed

AU - Bornholdt, Zachary A.

AU - Fusco, Marnie L.

AU - Abelson, Dafna M.

AU - Oda, Shunichiro

AU - Brown, Bethany L.

AU - Pham, Ha

AU - Rowland, Megan M.

AU - Agans, Krystle N.

AU - Geisbert, Joan B.

AU - Heinrich, Megan L.

AU - Kulakosky, Peter C.

AU - Shaffer, Jeffrey G.

AU - Schieffelin, John S.

AU - Kargbo, Brima

AU - Gbetuwa, Momoh

AU - Gevao, Sahr M.

AU - Wilson, Russell B.

AU - Saphire, Erica Ollmann

AU - Pitts, Kelly R.

AU - Khan, Sheik Humarr

AU - Grant, Donald S.

AU - Geisbert, Thomas

AU - Branco, Luis M.

AU - Garry, Robert F.

PY - 2016/10/15

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N2 - Background. Ebola virus disease (EVD) is a severe viral illness caused by Ebola virus (EBOV). The 2013-2016 EVD outbreak in West 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 × 105-9.0 × 108 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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KW - Diagnostic

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