Ebola Virus Diagnostics: The US Centers for Disease Control and Prevention Laboratory in Sierra Leone, August 2014 to March 2015

Flint, Mike; Goodman, Christin H.; Bearden, Scott W.; Blau, Dianna M.; Amman, Brian R.; Basile, Alison Jane; Belser, Jessica A.; Bergeron, Éric; Bowen, Michael D.; Brault, Aaron C.; Campbell, Shelley; Chakrabarti, Ayan K.; Dodd, Kimberly A.; Erickson, Bobbie R.; Freeman, Molly M.; Gibbons, Aridth; Guerrero, Lisa Wiggleton; Klena, John D.; Lash, R. Ryan; Lo, Michael K.; McMullan, Laura K.; Gbetuwa, Momoh; Massally, James L.B.; Goba, Augustine; Paddock, Christopher D.; Priestley, Rachael A.; Pyle, Meredith; Rayfield, Mark; Russell, Brandy J.; Salzer, Johanna S.; Sanchez, Angela J.; Schuh, Amy J.; Sealy, Tara K.; Steinau, Martin; Stoddard, Robyn A.; Taboy, Céline H.; Turnsek, Maryann; Wang, David; Zemtsova, Galina E.; Zivcec, Marko; Spiropoulou, Christina F.; Ströher, Ute; Towner, Jonathan S.; Nichol, Stuart T.; Bird, Brian H.

doi:10.1093/infdis/jiv361

Cited by 31 publications

(38 citation statements)

References 9 publications

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“…In October 2014, the laboratory moved to the Médecins Sans Frontiéres (MSF) Ebola Treatment Center (ETC) in Bo, Sierra Leone’s second largest city. As a result of the new location with improved accessibility and a simple laboratory set-up, approximately 26,000 diagnostic specimens were processed over 14 months [2]. The large number of specimens tested employing the same VSPB EBOV real-time reverse transcription polymerase chain reaction (qRT-PCR) assays allowed for analysis of assay robustness and comparison of blood and oral swab specimen types leading to enhanced result interpretation.…”

Section: Introductionmentioning

confidence: 99%

Ebola Virus Disease Diagnostics, Sierra Leone: Analysis of Real-time Reverse Transcription–Polymerase Chain Reaction Values for Clinical Blood and Oral Swab Specimens

et al. 2016

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During the Ebola virus outbreak of 2013–2016, the Viral Special Pathogens Branch field laboratory in Sierra Leone tested approximately 26,000 specimens between August 2014 and October 2015. Analysis of the B2M endogenous control Ct values showed its utility in monitoring specimen quality, comparing results with different specimen types, and interpretation of results. For live patients, blood is the most sensitive specimen type and oral swabs have little diagnostic utility. However, swabs are highly sensitive for diagnostic testing of corpses.

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Section: Introductionmentioning

confidence: 99%

Ebola Virus Disease Diagnostics, Sierra Leone: Analysis of Real-time Reverse Transcription–Polymerase Chain Reaction Values for Clinical Blood and Oral Swab Specimens

et al. 2016

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“…The steps in the QIAamp viral RNA extraction method from Qiagen that was used during the recent outbreak (15) are (i) sample collection; (ii) triple packing systems (5) for the shipment and transport of samples to high-containment laboratories (16); (iii) pipetting of aliquots; (iv) addition of AVL buffer; (v) incubation; (vi) addition of ethanol; and (vii) disinfection using 0.5% hypochlorite for 5 min before release from the glove box (17). These handling steps can be eliminated if efficient bedside inactivation of EBOV is obtained.…”

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confidence: 99%

Rapid Bedside Inactivation of Ebola Virus for Safe Nucleic Acid Tests

et al. 2016

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Rapid bedside inactivation of Ebola virus would be a solution for the safety of medical and technical staff, risk containment, sample transport, and high-throughput or rapid diagnostic testing during an outbreak. We show that the commercially available Magna Pure lysis/binding buffer used for nucleic acid extraction inactivates Ebola virus. A rapid bedside inactivation method for nucleic acid tests is obtained by simply adding Magna Pure lysis/binding buffer directly into vacuum blood collection EDTA tubes using a thin needle and syringe prior to sampling. The ready-to-use inactivation vacuum tubes are stable for more than 4 months, and Ebola virus RNA is preserved in the Magna Pure lysis/binding buffer for at least 5 weeks independent of the storage temperature. We also show that Ebola virus RNA can be manually extracted from Magna Pure lysis/binding buffer-inactivated samples using the QIAamp viral RNA minikit. We present an easy and convenient method for bedside inactivation using available blood collection vacuum tubes and reagents. We propose to use this simple method for fast, safe, and easy bedside inactivation of Ebola virus for safe transport and routine nucleic acid detection. The most recent Ebola virus disease (EVD) outbreak began in West Africa in December 2013. As of March 2016, the number of confirmed, probable, and suspected EVD cases reported worldwide was 28,646. Guinea, Liberia, and Sierra Leone were the most affected countries with 3,804, 10,666 and 14,122 cases, respectively (1).Ebola virus (EBOV) is classified as a risk group 4 pathogen that requires handling under biosafety level 4 (BSL-4) conditions. To meet this requirement, several mobile BSL-4 facilities were used during the recent West Africa outbreak (1, 2). However, extensive safety precautions and training of medical and technical staff are needed to ensure personal safety (2-6). As of August 2015, 880 health care workers had been diagnosed with EVD, and 512 had died from the disease (7). Rapid bedside inactivation of EBOV would be a solution for the safety of medical and technical staff, risk containment, and easier transport of samples without requiring expensive category A shipping. Additionally, this process removes the need for sample handling under high-containment environments and facilitates high-throughput and rapid testing under nonbiosafety laboratory conditions and, thus, a rapid diagnosis of the disease.There is a need for a simple, efficient, and safe bedside inactivation method for EBOV. Presently, laboratory EBOV inactivation is accomplished by gamma irradiation (8), UV radiation (9), nanoemulsion (10), and photoinducible alkylating agents (11), but these methods are not applicable in outbreak situations or as bedside inactivation methods. Other EBOV inactivation methods, such as acetic acid (12), heat (12), AVL buffer (13), TRIzol (13) or the combination of heat and Triton X-100 (14), are more applicable in outbreak situations and are currently used in field laboratories. Unfortunately, all of these methods require...

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“…Per the FDA authorizations (64)(65)(66), use of the U.S. CDC and DoD assays is restricted to facilities designated by these agencies; thus, these assays are not commercially available to clinical laboratories. Deployment of the CDC NP and VP40 assays in a field laboratory in Sierra Leone was recently described (68), although an evaluation of assay performance in this setting has not been provided. Data regarding clinical performance of the EZ1 and LightMix assays are not available.…”

Section: Evd Diagnostic Tests With Emergency Use Authorizationmentioning

confidence: 99%

Diagnosis of Ebola Virus Disease: Past, Present, and Future

2016

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SUMMARYLaboratory diagnosis of Ebola virus disease plays a critical role in outbreak response efforts; however, establishing safe and expeditious testing strategies for this high-biosafety-level pathogen in resource-poor environments remains extremely challenging. Since the discovery of Ebola virus in 1976 via traditional viral culture techniques and electron microscopy, diagnostic methodologies have trended toward faster, more accurate molecular assays. Importantly, technological advances have been paired with increasing efforts to support decentralized diagnostic testing capacity that can be deployed at or near the point of patient care. The unprecedented scope of the 2014-2015 West Africa Ebola epidemic spurred tremendous innovation in this arena, and a variety of new diagnostic platforms that have the potential both to immediately improve ongoing surveillance efforts in West Africa and to transform future outbreak responses have reached the field. In this review, we describe the evolution of Ebola virus disease diagnostic testing and efforts to deploy field diagnostic laboratories in prior outbreaks. We then explore the diagnostic challenges pervading the 2014-2015 epidemic and provide a comprehensive examination of novel diagnostic tests that are likely to address some of these challenges moving forward.

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Ebola Virus Diagnostics: The US Centers for Disease Control and Prevention Laboratory in Sierra Leone, August 2014 to March 2015

Cited by 31 publications

References 9 publications

Ebola Virus Disease Diagnostics, Sierra Leone: Analysis of Real-time Reverse Transcription–Polymerase Chain Reaction Values for Clinical Blood and Oral Swab Specimens

Ebola Virus Disease Diagnostics, Sierra Leone: Analysis of Real-time Reverse Transcription–Polymerase Chain Reaction Values for Clinical Blood and Oral Swab Specimens

Rapid Bedside Inactivation of Ebola Virus for Safe Nucleic Acid Tests

Diagnosis of Ebola Virus Disease: Past, Present, and Future

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