The document describes a prototype for a next generation rapid HIV test developed by OraSure Technologies. The prototype aims to detect HIV antibodies earlier than current rapid tests by achieving sensitivity similar to 3rd generation ELISA tests while maintaining a 10-minute test time. Performance studies showed the prototype detected HIV antibodies earlier than a FDA-approved rapid test in 50% of seroconversion panels tested, with an average of 5 days earlier detection. Specificity of the prototype was 99.87% in plasma samples. The prototype achieved all design goals of earlier detection of HIV antibodies during acute infection and a reduced test time.

1. Performance of a Prototype Rapid,
Point-of-Care Test* for Identifying
HIV Infected Individuals
OraSure Technologies, Inc.
2011 National HIV Prevention Conference
* In development. Not FDA approved
1

2. OraQuick® Rapid Test Platform
• OraQuick ADVANCE® Rapid HIV-1/2 Antibody Test
is widely used for rapid HIV testing
• Same test platform supports blood and oral fluid
specimens
• Enhanced version of the OraQuick HIV test was
released in early 2009
– Extended shelf life (30 months)
– Improved specificity1
• First FDA approved rapid HCV test uses the same
OraQuick test platform
2
1HIV Diagnostic Conference (2010)

3. Key Design Goals of a Next Generation
OraQuick® HIV Test
• Earlier detection of antibody in acute
infection
• Detection of antibody contemporaneous with 3rd
generation antibody EIAs
• Reduced assay time
• 10-minute test
• Retain simplicity and flexibility
• Low sample size (5uL) for whole blood
• Retain oral fluid capability
• Maintain high specificity
3

4. Defining the Target: Reducing the Time to
Detection of HIV Antibodies in Early Infection
Procleix (NAT) Time to Detection of HIV antibody (Days)
Rapid HIV Tests
WB Indeterminate
Lateral Flow
3rd Gen EIA
WB Positive
25 20 15 10 5 0 (Days)
Delay in time to detection between
rapid lateral flow tests and 3rd Gen EIA
4
Adapted from: Owen SM et al. (2008) J. Clin Microbiol. 46; 1588-1595

5. Prototype Next Generation OraQuick®
HIV Test*
Test Device
Developer Reusable
Vial Test Stand
Solution
Absorbent
Packet
Specimen
Collection
Loop
5
*In development. Not FDA approved

6. Prototype Next Generation OraQuick®
HIV Test: Device Design
• Intended Use
– Single-use, qualitative immunoassay
– HIV-1 and HIV-2 antibody detection
– Compatible with whole blood, oral fluid, serum and plasma
• Test Design
– Use of combination of HIV recombinant antigens and
synthetic peptides
– Novel biochemistry allows for improved reaction kinetics and
high affinity antibody capture
– Improved seroconversion sensitivity and improved analytical
sensitivity
6

7. Seroconversion Sensitivity of Prototype Next
Gen OraQuick® HIV Test vs. 3rd Generation EIA
Average
Number Average
Number Time to Delay Days
Number of Number of Detected Time to
Detected Detection (OQ HIV Next Gen
Panels Concordant Earlier by Detection
Earlier by OQ HIV -EIA)
Tested Series OQ HIV by EIA
EIA* Next Gen (95% CIs)
Next Gen (Days)
(Days)
0.54
24 14 7 3 32.21 32.75
(-0.93 – 2.01)
*FDA approved 3rd Generation EIA
7

8. Comparison of Seroconversion Sensitivity
vs FDA Approved Rapid Test
OQ HIV
Rapid Test A HIV-1/2/O
• 24 seroconversion panels tested by Panel Next Gen
(15 min) EIA
(10 min)
prototype OraQuick Next Gen Test 916 30 30 30
and currently approved rapid test 925 44 44 44
926 27 27 27
(Test A) 930 7 10 7
• 12/24 (50%) panels detected earlier 933 21 27 21
934 7 7 0
by the prototype OraQuick test 943 19 21 19
• 4 series did not seroconvert in 947 9 11 9
952 17 17 14
available bleeds in the FDA 957 23 28 28
approved rapid test 959 14 14 9
6243 32 32 32
• In the other 8 panels detected 9012 21 DNS 21
9015 35 DNS 35
earlier by the prototype Next Gen 9019 38 38 38
Test, the average difference in time 9021 57 DNS 57
to detection was 5 days 9032 29 38 36
9076 69 74 60
9077 52 52 52
9079 47 55 49
9081 26 26 24
9089 20 DNS 24
9096 18 18 18
12007 124 124 119
DNS = Did Not Seroconvert
8

9. Performance with Commercial Low Titer Panel PRB107
US FDA-Licensed Anti-HIV
OraQuick Confirmatory Test
HIV-1/2 HIV Ag
HIV Next
EIA EIA
Gen
Western Blot
10-min Banding
Specimen # s/co Result s/co
read Pattern
107-1 R 6.6 No Bands Neg 5.1
107-2 R 3.7 24, 55 Ind 14.0
107-3 R 4.5 No Bands Neg 0.8
107-4 R 9.7 No Bands Neg 1.2
107-5 NR 0.1 No Bands Neg 0.4
107-6 R 7.3 No Bands Neg 4.4
107-7 R 1.0 No Bands Neg 0.4
107-8 R 7.1 No Bands Neg 19.3
107-9 NR 2.4 No Bands Neg >29.9
107-10 R 3.5 No Bands Neg 24.5
107-11 R 3.3 24, 55, 160 Pos 3.9
107-12 NR 5.0 No Bands Neg 11.3
107-13 R 3.3 24 Ind 0.8
107-14 R 14.3 24, 160 Pos 1.7
107-15 R 2.7 24 Ind 0.8
9

10. Performance with Commercial Low Titer Panel PRB108
US FDA-Licensed Anti-HIV
Confirmatory Test
OQ HIV Rapid HIV-1/2
Next Gen Test “A” EIA
Western Blot
10-min 15-min
Specimen # s/co Banding Pattern Result
read read
108-1 R R 10.4 24, 55, 160 Pos
108-2 NR NR 0.2 No Bands Neg
108-3 R R 9.3 160 Ind
108-4 R R 15.7 41, 120, 160 Pos
108-5 R R 4.8 24, 41, 55, 120, 160 Pos
108-6 R R 6.4 160 Ind
108-7 R R 7.3 24, 160 Pos
108-8 R R 11.0 24, 160 Pos
108-9 R R 11.8 24, 160 Pos
108-10 R R 9.6 24 Ind
108-11 R R 10.1 24, 160 Pos
108-12 R NR 2.8 No Bands Neg
108-13 R NR 13.1 24 Ind
108-14 R NR 9.6 No Bands Neg
108-15 R R 16.2 160 Ind
10

11. Genotype Performance with Worldwide Panel
US FDA-Licensed Anti-HIV Confirmatory Test
OraQuick
HIV-1/2
HIV Next
EIA
Gen
HIV-1 Western Blot
10-min
Specimen # Origin Genotype s/co Banding Pattern Result
read
303-1 Ghana A R >15.9 18, 24, 31, 41, 51, 55, 66, 120, 160 Pos
303-2 Ghana A R >15.9 18, 24, 31, 41, 51, 55, 66, 120, 160 Pos
303-3 South Africa C R >15.9 18, 24, 31, 41, 51, 55, 66, 120, 160 Pos
303-4 South Africa C R >15.9 18, 24, 41, 51, 55, 66, 120, 160 Pos
303-5 Uganda D R >15.9 18, 24, 31, 41, 51, 55, 66, 120, 160 Pos
303-6 Uganda D R >15.9 18, 24, 31, 41, 51, 55, 66, 120, 160 Pos
303-7 Ghana G R >15.9 18, 24, 31, 41, 51, 55, 66, 120, 160 Pos
303-8 Ivory Coast G R >15.9 18, 24, 31, 41, 51, 55, 66, 120, 160 Pos
303-9 Spain O R 2.4 31, 51, 66, 160 Ind
303-10 Argentina F R >15.9 18, 24, 31, 41, 51, 66, 120, 160 Pos
303-11 Ivory Coast HIV-2 R 0.3 24 Ind
303-12 Argentina Neg NR 0.4 No Bands Neg
303-13 Uganda A R >15.9 18, 24, 31, 41, 51, 55, 66, 120, 160 Pos
303-14 Zimbabwe C R >15.9 18, 24, 31, 41, 51, 55, 66, 120, 160 Pos
303-15 USA B R >15.9 18, 24, 31, 41, 51, 55, 66, 120, 160 Pos
11

12. HIV-2 Sensitivity
HIV-2 EIA
+ -
+ 171 0
Prototype
Next Gen
OraQuick
- 0 0
12

13. Specificity of Prototype OraQuick Next
Gen Test (Plasma)
HIV-1/2/O
EIA
+ -
+ 0 1
Prototype
Next Gen
OraQuick
- 0 749
99.87% specificity
(99.26% - 100%)
13

14. Performance in Preclinical Human
Subject Testing
Prospective Testing of 198 Low Risk Subjects
Specimen
Fingerstick Oral Fluid
Type
100% 99.50%
Specificity
(98.15% - 100%) (97.22% - 99.99%)
14

15. Conclusions
• Design goals of the Next Generation OraQuick HIV test have
been achieved
• Earlier detection of antibody in seroconversion
– Sensitivity similar to 3rd generation EIA
– Improved detection in early HIV infection compared to current
lateral flow rapid tests
– Measured difference in time to detection of antibody between EIA
and current rapid tests appears smaller than previously reported
• Shortened testing time – 10 minute prototype
• Retains oral fluid and blood capability
• Public health value of rapid HIV tests continues to reside in
identification of established, undiagnosed infection and the
immediacy of test results
15