Ebola Virus Disease (EVD)Laboratory Diagnostics
Dr Joanne O’GormanDeputy Director
National Virus Reference LaboratoryUniversity College Dublin
Ebola Conference 12.11.14 JOG
National Virus Reference Laboratory(NVRL)
• Located on campus in UCD
• Biosafety level 3 Facility(BSL-3)
• Member of ENIVD– European Network for Diagnostics
of “Imported” Viral Diseases
• Website http://nvrl.ucd.ie
Ebola Conference 12.11.14 JOG
Ebola Virus DiseaseLaboratory Criteria Case Defintion
Any of the following:
Isolation of Ebola virus from a clinical specimen.✖
Ebola Conference 12.11.14 JOG
Ebola Virus DiseaseLaboratory Criteria Case Defintion
Any of the following:
Isolation of Ebola virus from a clinical specimen.✖
Detection of Ebola virus nucleic acid in a clinicalspecimen and confirmation by sequencing or a
second assay on different genomic targets.✔
Ebola Conference 12.11.14 JOG
RealStar RT-PCR [Altona] **
In house RT-PCRIn house RT-PCR
Testing Methodology
5mls
Ebola Conference 12.11.14 JOG
Requesting a test for EVD
NVRL Contact Details Tel: 01 7164401Mob: 087 9806448
Ebola Conference 12.11.14 JOG
Requesting a test for EVD
Contact Details
Tel: 01 7164401Mob: 087 9806448
Ebola Conference 12.11.14 JOG
Packaging & TransportCategory A InfectiousSubstances• Category A infectious substances
are capable of causing permanentdisability, life threatening or fataldisease to humans or animalswhen exposure to them occurs.
• Category A infectious substanceshave two shipping names:“Infectious substances, Affectinghumans” (UN 2814) or“Infectious substances, affectinganimals” (UN 2900)
• Designated courier
Category A InfectiousSubstances• Category A infectious substances
are capable of causing permanentdisability, life threatening or fataldisease to humans or animalswhen exposure to them occurs.
• Category A infectious substanceshave two shipping names:“Infectious substances, Affectinghumans” (UN 2814) or“Infectious substances, affectinganimals” (UN 2900)
• Designated courier
Category B InfectiousSubstances
• Category B infectious substancesare infectious but do not meet thecriteria for Category A.
• Category B infectious substanceshave the proper shipping name“Biological Substance, CategoryB” and the identification numberUN 3373.
Ebola Conference 12.11.14 JOG
Main corridor
BL3 laboratory-50Pa
ClassI/ IIIMSC
ClassIIIMSC
Shower-10Pa
Outer lobby
Inner lobby-35Pa
-20oC -80oCCO2Inc. 40C
+10Pa18 ac/h
Pass throughautoclave
Containment envelope
NVRL Testing Process – BSL 3
Ebola Conference 12.11.14 JOG
A. Security & accessB. Air handlingC. Dis-infection & disposal
proceduresD. Protective equipment &
procedures
Laboratory & StaffPreparation (2)
Laboratory & StaffPreparation (2)
Specimen Preparation& Centrifugation
Specimen Preparation& Centrifugation
RNA ExtractionGuanidine thiocyanate.
RNA ExtractionGuanidine thiocyanate.
PCRPCR
Result Interpretation &
CommunicationResult Interpretation &
Communication
B
S
L
3
B
S
L
3
B
S
L
2
B
S
L
2
Ebola Conference 12.11.14 JOG
Frequently Asked Questions
1. Turn around time?
Most cases approx. 6 hours from receipt of samplein the NVRL.
Ebola Conference 12.11.14 JOG
Frequently Asked Questions
1 Turn Around Time?2 Sensitivity?
– Assays• Performance & Proficiency Testing
– Sample• EDTA• Within 12 hours
– Patient• Decision to restest and timing of retesting based on exposure risk
assessment and clinical features
Ebola Conference 12.11.14 JOG
Frequently Asked Questions
1 Turn Around Time ?2 Sensitivity?3 Specificity?
Ebola Conference 12.11.14 JOG
Frequently Asked Questions
1 Turn Around Time ?2 Sensitivity?3 Specificity?
• Two assays with two different targets• 15 patients tested to date – ALL NEGATIVE
Ebola Conference 12.11.14 JOG
Key Messages
• Refer to the Ebola Virus Disease Risk AssessmentAlgorithm available @hpsc.ie
• Contact the NVRL prior to sending a sample.
• Results will be communicated to the designatedperson as soon as they are available ( approx 6hours)
Ebola Conference 12.11.14 JOG
Acknowledgements
• NVRL staff– Margaret Duffy– Grainne Tuite– Jeff Connell– Suzie Coughlan– Ger Franzoni– Brian Keogan
http://nvrl.ucd.ieEbola Conference 12.11.14 JOG