Molecular Diagnostic Field Test for Point-of-Care Detection of Ebola Virus Directly from Blood

Jason W. Benzine, Kerry M. Brown, Krystle N. Agans, Ronald Godiska, Chad Mire, Krishne Gowda, Brandon Converse, Thomas Geisbert, David A. Mead, Yogesh Chander

Research output: Contribution to journalArticle

14 Scopus citations

Abstract

A molecular diagnostic method for robust detection of Ebola virus (EBOV) at the point of care (POC) directly from blood samples is described. This assay is based on reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) of the glycoprotein gene of EBOV. Complete reaction formulations were lyophilized in 0.2-mL polymerase chain reaction tubes. RT-LAMP reactions were performed on a battery-operated isothermal instrument. Limit of detection of this RT-LAMP assay was 2.8 × 102 plaque-forming units (PFU)/test and 1 × 103 PFU/test within 40 minutes for EBOV-Kikwit and EBOV-Makona, respectively. This assay was found to be specific for the detection of EBOV, as no nonspecific amplification was detected in blood samples spiked with closely related viruses and other pathogens. These results showed that this diagnostic test can be used at the point of care for rapid and specific detection of EBOV directly from blood with high sensitivity within 40 minutes.

Original languageEnglish (US)
Pages (from-to)S234-S242
JournalJournal of Infectious Diseases
Volume214
DOIs
StatePublished - Oct 15 2016

    Fingerprint

Keywords

  • Diagnostic test
  • Ebola virus
  • Isothermal amplification
  • Point of care
  • RT-LAMP

ASJC Scopus subject areas

  • Immunology and Allergy
  • Infectious Diseases

Cite this

Benzine, J. W., Brown, K. M., Agans, K. N., Godiska, R., Mire, C., Gowda, K., Converse, B., Geisbert, T., Mead, D. A., & Chander, Y. (2016). Molecular Diagnostic Field Test for Point-of-Care Detection of Ebola Virus Directly from Blood. Journal of Infectious Diseases, 214, S234-S242. https://doi.org/10.1093/infdis/jiw330