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

Testing Methodology

Zaire*SudanTai ForestBundibugyoRestonVirus*

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RealStar RT-PCR [Altona] **

In house RT-PCRIn house RT-PCR

Testing Methodology

5mls

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Requesting a test for EVD

Decision

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Requesting a test for EVD

Sample Types EDTA & Serum (WCB)

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

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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.

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Transport of Biological Material:UN2814

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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

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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

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Frequently Asked Questions

1. Turn Around Time ?

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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?

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?

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Frequently Asked Questions

1 Turn Around Time ?2 Sensitivity?3 Specificity?

• Two assays with two different targets• 15 patients tested to date – ALL NEGATIVE

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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)

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Acknowledgements

• NVRL staff– Margaret Duffy– Grainne Tuite– Jeff Connell– Suzie Coughlan– Ger Franzoni– Brian Keogan

http://nvrl.ucd.ieEbola Conference 12.11.14 JOG

Ebola Conference 12.11.14 JOG