Characterization and analytical validation of a new antigenic rapid diagnostic test for Ebola virus disease detection

Abstract

Hemorrhagic fever outbreaks are difficult to diagnose and control in part because of a lack of low-cost and easily accessible diagnostic structures in countries where etiologic agents are present. Furthermore, initial clinical symptoms are common and shared with other endemic diseases such as malaria or typhoid fever. Current molecular diagnostic methods such as polymerase chain reaction require trained personnel and laboratory infrastructure, hindering diagnostics at the point of need, particularly in outbreak settings. Therefore, rapid diagnostic tests such as lateral flow can be broadly deployed and are typically well-suited to rapidly diagnose hemorrhagic fever viruses, such as Ebola virus. Early detection and control of Ebola outbreaks require simple, easy-to-use assays that can detect very low amount of virus in blood. Here, we developed and characterized an immunoassay test based on immunochromatography coupled to silver amplification technology to detect the secreted glycoprotein of EBOV. The glycoprotein is among the first viral proteins to be detected in blood. This strategy aims at identifying infected patients early following onset of symptoms by detecting low amount of sGP protein in blood samples. The limit of detection achieved by this sGP-targeted kit is 2.2 x 10 4 genome copies/ml in plasma as assayed in a monkey analytical cohort. Clinical performance evaluation showed a specificity of 100% and a sensitivity of 85.7% when evaluated with plasma samples from healthy controls and patients infected with Zaire Ebola virus from Macenta, Guinea. This rapid and accurate diagnostic test could therefore be used in endemic countries for early detection of infected individuals in point of care settings. Moreover, it could also support efficient clinical triage in hospitals or clinical

Development of an Enzyme-Linked Immunosorbent Assay to Determine the Expression Dynamics of Ebola Virus Soluble Glycoprotein during Infection

“…Several other EBOV rapid diagnostic tests were generated and some of them were approved during the outbreak [ 28 ]. A recent study reported generation of an sGP and GP detection kit based on immunochromatography coupled with silver amplification technology [ 11 ]. The limit of detection for this assay was 2.21 × 10 4 genome copies/mL in plasma samples from an infected NHP.…”

“…The remaining 20–30% are left for the edited 8A full-length GP which is expressed on the virion surface, and the edited 9A small sGP (ssGP) [ 9 , 10 ]. Even though sGP is transcribed and expressed at a higher level compared to GP and is detected in the blood of EBOV-infected animals and humans, only few diagnostic tests targeting sGP exist [ 11 , 12 , 13 ].…”

Application of carbon nanomaterials in human virus detection

“…Facile, facilitated tests are required to detect and manage the Ebola endemics earlier tests which are capable of detecting the virus in blood at highly little quantities. The WHO announced rapid, sensitive, safe, and simple EBOV diagnostic assays in November 2014 [ [65] , [66] , [67] ]. Wen et al.…”

COVID-19 Compared with Other Viral Diseases: Novelties, Progress, and Challenges

“…Ebola Viral Disease (EVD) is a severe and overwhelmingly fatal disease [30]; death rate is approximately 80% among those infected. It is caused by the EBOV, a member of the filovirus family and occurs in humans and other primates [31]. An EBOV is a single-stranded, non-segmented, and negativesense RNA virus.…”

“…EBOVs are thought to be zoonotic; however, their original reservoir has yet to be confirmed. African fruit bats are regarded as the most likely natural reservoir for EBOVs [31,33]. Initial human infection with Ebola virus occurs through contact with an infected animal, such as a fruit bat or other non-human primate.…”