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

• Published in: The Journal of infectious diseases 2016-10, Vol.214 (suppl 3), p.S234-S242
• Main Authors: Benzine, Jason W., Brown, Kerry M., Agans, Krystle N., Godiska, Ronald, Mire, Chad E., Gowda, Krishne, Converse, Brandon, Geisbert, Thomas W., Mead, David A., Chander, Yogesh
• Format: Article
• Language: English
• Subjects: DIAGNOSIS; Ebola Outbreak in West Africa; Ebolavirus - genetics; Ebolavirus - isolation & purification; Hemorrhagic Fever, Ebola - diagnosis; Hemorrhagic Fever, Ebola - virology; Humans; Molecular Diagnostic Techniques; Nucleic Acid Amplification Techniques - methods; Point-of-Care Systems; RNA, Viral - blood; RNA, Viral - genetics; Sensitivity and Specificity
• ISSN: 0022-1899; 1537-6613
• DOI: 10.1093/infdis/jiw330

A molecular diagnostic method for robust detection of Ebola virus (EBOV) at the point of care 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 × 10 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.