Experiences of outbreak laboratory management in the Ebola Disease outbreak in West-Africa 2014-2015

During the Ebola Disease outbreak in 2014-2015 West-Africa about 24 organizations operated laboratories at 40 sites in Guinea, Sierra Leone and Liberia. Representatives of ten organisations which had deployed laboratories to 16 sites across the three countries in West-Africa convened for a two day symposium in Dakar (4-5.02.16) to exchange their experiences. This article summarizes the discussion and points made during the discussion of the laboratory deployment experiences during the epidemic touching organisational and procedural issues.Additional co-authors: P Jansen van Vuren, K Stroecker, J Paweska, C Picard, H Sheeley, P Smit, AA Sal

South African Ebola diagnostic response in Sierra Leone : a modular high biosafety field laboratory

BACKGROUND : In August 2014, the National Institute for Communicable Diseases (NICD) in South Africa established a modular high-biosafety field Ebola diagnostic laboratory (SA FEDL) near Freetown, Sierra Leone in response to the rapidly increasing number of Ebola virus disease (EVD) cases. METHODS AND FINDINGS : The SA FEDL operated in the Western Area of Sierra Leone, which remained a ªhotspotº of the EVD epidemic for months. The FEDL was the only diagnostic capacity available to respond to the overwhelming demand for rapid EVD laboratory diagnosis for several weeks in the initial stages of the EVD crisis in the capital of Sierra Leone. Furthermore, the NICD set out to establish local capacity amongst Sierra Leonean nationals in all aspects of the FEDL functions from the outset. This led to the successful hand-over of the FEDL to the Sierra Leone Ministry of Health and Sanitation in March 2015. Between 25 August 2014 and 22 June 2016, the laboratory tested 11,250 specimens mostly from the Western Urban and Western Rural regions of Sierra Leone, of which 2,379 (21.14%) tested positive for Ebola virus RNA. CONCLUSIONS : he bio-safety standards and the portability of the SA FEDL, offered a cost-effective and practical alternative for the rapid deployment of a field-operated high biocontainment facility. The SA FEDL teams demonstrated that it is highly beneficial to train the national staff in the course of formidable disease outbreak and accomplished their full integration into all operational and diagnostic aspects of the laboratory. This initiative contributed to the international efforts in bringing the EVD outbreak under control in Sierra Leone, as well as capacitating local African scientists and technologists to respond to diagnostic needs that might be required in future outbreaks of highly contagious pathogens.S1 Video. ªHotº processing of Ebola clinical specimens, PPE and decontamination procedures in South African modular, field-operated biocontainment facility in Sierra Leone.Janusz T Paweska was supported by funding from National Research Foundation and the Global Disease Detection Programmehttp://www.plosntds.orgam2017Microbiology and Plant Patholog

Comparative Evaluation of the Diagnostic Performance of the Prototype Cepheid GeneXpert Ebola Assay

The Ebola virus disease (EVD) outbreak in West Africa has highlighted an urgent need for point-of-care (POC) assays for the diagnosis of this devastating disease in resource-limited African countries. The diagnostic performance characteristics of a prototype Cepheid GeneXpert Ebola POC used to detect Ebola virus (EBOV) in stored serum and plasma samples collected from suspected EVD cases in Sierra Leone in 2014 and 2015 was evaluated. The GeneXpert Ebola POC is a self-contained single-cartridge automated system that targets the glycoprotein (GP) and nucleoprotein (NP) genes of EBOV and yields results within 90 min. Results from 281 patient samples were compared to the results of a TaqMan real-time reverse transcription-PCR (RT-PCR) targeting the polymerase gene and performed on two real-time PCR machines. Agreement between the three platforms was 100% at cycle threshold (CT) values of<34.99, but discordant results were noted between CT values of 35 and 45.The diagnostic sensitivity of the three platforms was 100% in 91 patient samples that were confirmed to be infectious by virus isolation. All three molecular platforms detected viral EBOV RNA in additional samples that did not contain viable EBOV. The analytical sensitivity of the GeneXpert Ebola POC for the detection of NP was higher, and comparable to that of polymerase gene detection, than that for the detection of GP when using a titrated laboratory stock of EBOV. There was no detectable cross-reactivity with other hemorrhagic fever viruses or arboviruses. The GeneXpert Ebola POC offers an easy to operate and sensitive diagnostic tool that can be used for the rapid screening of suspected EVD cases in treatment or in holding centers during EVD outbreaks.NICD was supported by Cepheid by provision of GeneXpert® Ebola POC IV system, GeneXpert® Dx software package, disposable prototype GeneXpert® Ebola cartridges, and funds for covering costs of running polymerase (L) gene TaqMan real-time RT-PCR assays.http://jcm.asm.org2016-08-31hb2016Microbiology and Plant Patholog

Comparative evaluation of the diagnostic performance of the prototype Cepheid GeneXpert Ebola assay

The Ebola virus disease (EVD) outbreak in West Africa has highlighted an urgent need for point-of-care (POC) assays for the diagnosis of this devastating disease in resource-limited African countries. The diagnostic performance characteristics of a prototype Cepheid GeneXpert Ebola POC used to detect Ebola virus (EBOV) in stored serum and plasma samples collected from suspected EVD cases in Sierra Leone in 2014 and 2015 was evaluated. The GeneXpert Ebola POC is a self-contained single-cartridge automated system that targets the glycoprotein (GP) and nucleoprotein (NP) genes of EBOV and yields results within 90 min. Results from 281 patient samples were compared to the results of a TaqMan real-time reverse transcription-PCR (RT-PCR) targeting the polymerase gene and performed on two real-time PCR machines. Agreement between the three platforms was 100% at cycle threshold (CT) values of<34.99, but discordant results were noted between CT values of 35 and 45.The diagnostic sensitivity of the three platforms was 100% in 91 patient samples that were confirmed to be infectious by virus isolation. All three molecular platforms detected viral EBOV RNA in additional samples that did not contain viable EBOV. The analytical sensitivity of the GeneXpert Ebola POC for the detection of NP was higher, and comparable to that of polymerase gene detection, than that for the detection of GP when using a titrated laboratory stock of EBOV. There was no detectable cross-reactivity with other hemorrhagic fever viruses or arboviruses. The GeneXpert Ebola POC offers an easy to operate and sensitive diagnostic tool that can be used for the rapid screening of suspected EVD cases in treatment or in holding centers during EVD outbreaks.NICD was supported by Cepheid by provision of GeneXpert® Ebola POC IV system, GeneXpert® Dx software package, disposable prototype GeneXpert® Ebola cartridges, and funds for covering costs of running polymerase (L) gene TaqMan real-time RT-PCR assays.http://jcm.asm.org2016-08-31hb2016Microbiology and Plant Patholog

Layout of the SA FEDL in Freetown-Lakka, Sierra Leone with emergency generator and wiring to allow for rapid switch to generator mode in case of power failure.

<p>(<b>A</b>) Biocontainment negative pressure chamber (IsoArk), (<b>B</b>) Room housing biocontainment negative pressure chamber, (<b>C</b>) Donning room, (<b>D</b>) Doffing room, (<b>E</b>) Laboratory airlock area, (<b>F</b>) PCR amplification room, (<b>G</b>) PCR master mix room, (<b>H</b>) Specimens and reagents storage area, (<b>I</b>) RNA extraction room, (<b>J</b>) Facility entrance, (<b>K</b>) Toilet, (<b>L</b>) Office 1, (<b>M</b>) Office 2, (<b>N</b>) Office 3. Petrol generator (5.5 kVa) placement indicated by the red rectangle, distribution of extension cords are indicated with red lines, and emergency connection points by red stars.</p

South African Ebola diagnostic response in Sierra Leone: A modular high biosafety field laboratory - Fig 3

<p>(<b>A</b>) Operators dressed in BSL3 PPE processing clinical specimens from EVD suspected cases in a glovebox located within the IsoArk biocontainment negative pressure chamber, (<b>B</b>) Blood tubes in centrifuge adapters with safety caps passed into the main chamber of the glovebox for “hot” inactivation and aliquoting for long-term storage.</p

Ebola virus polymerase gene TaqMan real-time RT-PCR results in buccal swabs.

<p>Ebola virus polymerase gene TaqMan real-time RT-PCR results in buccal swabs.</p

Results of EBOV L-gene RT-PCR in sera subjected to RNA manual (black dots) and automated (circles) extraction.

<p>Results of EBOV L-gene RT-PCR in sera subjected to RNA manual (black dots) and automated (circles) extraction.</p

Ebola virus polymerase gene TaqMan real-time RT-PCR results in blood specimens.

<p>Ebola virus polymerase gene TaqMan real-time RT-PCR results in blood specimens.</p

Operators dressed in BSL3 PPE (scrubs, Tyvek suits, surgical gown, double pair surgical gloves, gumboots, overshoes and PAPR with full face hood) entering the IsoArk main chamber through the airlock.

<p>Operators dressed in BSL3 PPE (scrubs, Tyvek suits, surgical gown, double pair surgical gloves, gumboots, overshoes and PAPR with full face hood) entering the IsoArk main chamber through the airlock.</p