Rapid Response Ebola Test (RRET)

Technical Report

Overview: The Ebola Pandemic So Far…

Scary Stats:

• Africa

• Europe

• America

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Country Total Cases Lab-confirmed

Cases

Total Deaths

Guinea 2283 2035 1412

Liberia 7719 2830 3177

Sierra Leone 7798 6317 1742

Mali 8 7 6

Senegal 1 1 0

Nigeria 20 19 8

Spain 1 1 0

United States 4 4 1

Total 17834 11214 6346

CDC – as at Dec. 9, 2014

Overview: Need for RRET

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• Rapid diagnostics

30mins T/A time

• Surveillance

Ports of entry and other health centres

RRET Features

• Rapid genetic diagnostic test • Based on the Loop-mediated

isothermal amplification (LAMP) technique

• Single isothermal temperature (630C)– Water bath or heating block

sufficient– No need for a thermal cycler or

PCR equipment

• Visual detection– Fluorescence at 240nm-260nm

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Comparison with gold standard - PCRSample Collection

Urine Sputum

RNA Extraction Phase(Kits from Quiagen, Illumina, etc.)

cDNA Synthesis and Amplification

RT-PCR

Gel Electrophoresis

RT-LAMP(RRET)

Gel Electrophoresis

Fluorescence Detection

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2mins

3mins – 1hr

Minimum of 2hrs

30mins

15 – 30mins

30mins (Optional)

1min

3mins – 1hr

2mins

RRET Validation Data: From current outbreak samples

CLARIFICATIONS

• RRET Primers target only– ZEBOV Strain

• PCR Wobble Primers target– Filo– Marburg– ZEBOV

• Positive Control RefSeq– PCR (GP Seq)– RRET (Trailer

Seq)

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S/N Sample ID

RT-PCR

RRET

1 061195

NEG NEG

2 061228

NEG NEG

3 061215

NEG NEG

4 061305

NEG NEG

5 061306

NEG NEG

6 061231

NEG NEG

7 061307

NEG NEG

8 061338

NEG NEG

S/N

Sample ID

RT-PCR

RRET

9 061337

NEG NEG

10 018 POS POS

11 MAINLAND 10

POS POS

12 061232

POS POS

13 061214

POS NEG

14 019 POS POS

15 PS POS POS

RRET Sensitivity/Specificity Values: With respect to gold standard

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SENSITIVITY 80%

SPECIFICITY 100%

POSITIVE PREDICTIVE VALUE 100%

NEGATIVE PREDICTIVE VALUE 90%

Sensitivity defined as probability that a test will be positive when disease is present.

Specificity defined as probability that a test will be negative when disease is absent.

Positive Predictive Value defined as probability that the disease is present when the test is positive.

Negative Predictive Value defined as probability that the disease is not present when test is negative.

Marketing and Deployment• Ports of entry

– Surveillance

• Secondary & tertiary health centres– General hospitals– Teaching hospitals– Specialist centres

• Other diagnostic labs– Universities/other educational institutions for student screening– Private/public sector institutions for staff screening

• Mobile diagnostics– Sports screening (ANC, 2015 @E.Guinea)– Supplies for volunteer clinicians at hot zones

RRET Accessories needed @staging area or mobile lab:

1. Water bath or a heating block.

2. Adjustable UV lamp or photo-transilluminator (wavelengths 240nm – 370nm).

3. Protective glass shield or eye googles.

4. Hand gloves.

5. Bio-harzard bin.