1. Rapid Response Ebola Test (RRET) Technical Report

2. Overview: The Ebola Pandemic So Far… Scary Stats: Africa Europe America 2 CountryTotal CasesLab-confirmed Cases Total Deaths Guinea228320351412 Liberia771928303177 Sierra Leone779863171742 Mali876 Senegal110 Nigeria20198 Spain110 United States441 Total17834112146346 CDC – as at Dec. 9, 2014

3. Overview: Need for RRET 3 Rapid diagnostics 30mins T/A time Surveillance Ports of entry and other health centres

4. RRET Features Rapid genetic diagnostic test Based on the Loop-mediated isothermal amplification (LAMP) technique Single isothermal temperature (63 0 C) – Water bath or heating block sufficient – No need for a thermal cycler or PCR equipment Visual detection – Fluorescence at 240nm- 260nm 4

5. Comparison with gold standard - PCR Sample Collection RNA Extraction Phase (Kits from Quiagen, Illumina, etc.) cDNA Synthesis and Amplification RT-PCR Gel Electrophoresis RT-LAMP (RRET) Gel Electrophoresis Fluorescence Detection Urine Sputum Blood Swab 5 2mins 3mins – 1hr Minimum of 2hrs 30mins 15 – 30mins 30mins (Optional) 1min 3mins – 1hr 2mins

6. 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) 6 S/NSample ID RT- PCR RRET 1061195NEG 2061228NEG 3061215NEG 4061305NEG 5061306NEG 6061231NEG 7061307NEG 8061338NEG S/NSample ID RT- PCR RRET 9061337NEG 10018POS 11MAINL AND 10 POS 12061232POS 13061214POSNEG 14019POS 15PSPOS

7. RRET Sensitivity/Specificity Values: With respect to gold standard 7 SENSITIVITY80% SPECIFICITY100% POSITIVE PREDICTIVE VALUE100% NEGATIVE PREDICTIVE VALUE90% 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.

8. 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

9. 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.