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 E. Mire; Krishne Gowda; Brandon Converse; Thomas W. Geisbert; David A. Mead; Yogesh Chander

doi:10.1093/infdis/jiw330

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 E. Mire, Krishne Gowda, Brandon Converse, Thomas W. Geisbert, David A. Mead, Yogesh Chander

Microbiology And Immunology

Research output: Contribution to journal › Article › peer-review

20 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 language	English (US)
Pages (from-to)	S234-S242
Journal	Journal of Infectious Diseases
Volume	214
DOIs	https://doi.org/10.1093/infdis/jiw330
State	Published - Oct 15 2016

Keywords

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

ASJC Scopus subject areas

Immunology and Allergy
Infectious Diseases

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10.1093/infdis/jiw330

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title = "Molecular Diagnostic Field Test for Point-of-Care Detection of Ebola Virus Directly from Blood",

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.",

keywords = "Diagnostic test, Ebola virus, Isothermal amplification, Point of care, RT-LAMP",

author = "Benzine, {Jason W.} and Brown, {Kerry M.} and Agans, {Krystle N.} and Ronald Godiska and Mire, {Chad E.} and Krishne Gowda and Brandon Converse and Geisbert, {Thomas W.} and Mead, {David A.} and Yogesh Chander",

note = "Publisher Copyright: {\textcopyright} 2016 The Author.",

year = "2016",

month = oct,

day = "15",

doi = "10.1093/infdis/jiw330",

language = "English (US)",

volume = "214",

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AU - Benzine, Jason W.

AU - Brown, Kerry M.

AU - Agans, Krystle N.

AU - Godiska, Ronald

AU - Mire, Chad E.

AU - Gowda, Krishne

AU - Converse, Brandon

AU - Geisbert, Thomas W.

AU - Mead, David A.

AU - Chander, Yogesh

PY - 2016/10/15

Y1 - 2016/10/15

N2 - 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.

AB - 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.

KW - Diagnostic test

KW - Ebola virus

KW - Isothermal amplification

KW - Point of care

KW - RT-LAMP

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ER -

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

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Keywords

ASJC Scopus subject areas

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