Transcript VITROS® HIV-1/2 Assay - 2016 HIV Diagnostics Conference

Development of an Anti-HIV 1+2
Assay for use on a Random Access
System
B Boyer, S Edwards, JM Glover: Ortho-Clinical Diagnostics,
Rochester NY
This assay is currently pending FDA approval.
This is not a solicitation for sale but a scientific
exchange of information for study.
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Development of an Anti-HIV 1+2 Assay for
use on a Random Access System
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Description of the test/protocol
Clinical Performance
Result Algorithm
Intellicheck ®
Summary/Conclusion
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The Vitros® Anti-HIV 1+2 Assay*
Test for the in vitro qualitative detection of antibodies to
Human Immunodeficiency Virus types 1 and/or 2 (anti-HIV 1
and anti-HIV 2) in human serum and plasma (heparin,
EDTA or citrate) using the VITROS ® ECi/ECiQ
Immunodiagnostic System.
* Pending FDA approval
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Architecture
• HIV 1 Env 13 - gp 120 and gp 41 region.
• HIV-1 Env 10 - gp41 region which extends beyond the Cterminus of Env 13.
• HIV-1 p24 - full length core protein of HIV-1.
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• HIV-2 Env AL - contains a region from gp 36 of HIV-2.
Protocol
• 80 l sample
• 20 l assay reagent
• Incubate 29 minutes 20
seconds at 37°C
• Wash
• 100 l conjugate reagent
• Incubate 8 minutes at
37°C
• Wash
• 200 l signal reagent
• Read
Assay Type
Immunometric assay
Assay Time and Temperature
Incubation time:
37 minutes
Time to first result:
48 minutes
Temperature:
37 °C
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Calibration and QC
• A single calibrator is run in duplicate once every 28 days
• 3 QC controls are available.
– Negative, HIV 1, HIV 2
– Recommendation is that these controls are run in singleton:
• once every 24 hours of testing
• after calibration
• after certain service events
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Performance: US Clinical Trials
High Risk Populations
Population
Positive Percent
Agreement
Negative Percent
Agreement
High Risk (U.S.)
100%
(54/54)
99.81%
(2117/2121)
High Risk
(Ivory Coast)
100%
(26/26)
98.92%
(457/462)
HIV Positive
(U.S.)
100%
(1121/1121)
HIV Positive
(International)
99.48%
(193/194)
HIV-2 Positive
(Ivory Coast)
100%
(208/208)
Pregnant
(High Risk–U.S.)
100%
(5/5)
99.59%
(243/244)
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Performance: US Clinical Trials (continued)
Low Risk Populations
Population
Pregnant
(Low Risk–U.S.)
Positive Percent
Agreement
Negative Percent
Agreement
100%
(1/1)
100%
(296/296)
Labor and Delivery (Low
Risk–U.S.)
Insurance Applicants
(Low Risk–U.S.)
97.96%
(48/49)
100%
(5/5)
Pediatric
(Low Risk–U.S.)
Pediatric HIV Positive
(U.S.)
Total (High and low
risk populations)
99.80%
(992/994)
100%
(99/99)
100%
(40/40)
99.94%
(1653/1654)
99.70%
(4252/4265)8
450
926
500
3274
Performance: US Clinical Trials (continued)
HIV Ab Positive
HIV Ab Negative
350
Borderline
Range
300
250
200
150
100
50
Result (Sample/Cut-off)
80
10
0
60
40
20
9
7
5
3
1
0.
8
0.
6
0.
4
0.
2
0
0
Number of samples
400
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Performance: Seroconversion
20 Seroconversion panels tested with a licensed assay and the
Vitros ® Assay*. 14 panels converted at the same sample. 6
panels showed differential conversion between assays.
Days to Evidence of HIV Infection
Panel ID
Licensed
Assay
Vitros ®
Assay*
-
+
-
+
Difference days to HIV
reactivity: Licensed assay
minus Vitros ® Assay*
PRB927
0
28
28
33
-5
PRB929
21
25
18
21
4
PRB934
0
7
0
0
7
PRB940
7
11
0
7
4
PRB952
14
17
10
14
3
PRB959
7
9
0
7
2
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* Pending FDA approval
HIV 1 Genotype Detection
Mean Result
Genotype
(s/c)
n
Result Range
(s/c)
A
64.2
3
55.3 to 82.6
B
80.7
3
74.1 to 86.7
C
72.3
2
55.2 to 89.4
D
64.4
4
59.2 to 73.0
E
75.6
2
72.7 to 78.4
F
79.6
1
79.6
G
66.7
4
56.4 to 85.3
O
15.3
14
2.73 to 36.6
100% Sensitivity
with all genotypes
tested – including O
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Performance: Precision
Clinical
Site
Site 1
Site 2
Site 3
Mean VITROS
Anti-HIV 12 Assay
Results (s/c)
0.07
6.06
4.12
1.32
0.08
6.66
4.39
1.39
0.07
6.22
4.42
1.34
Negative
HIV-1
HIV-2
HIV-1
Negative
HIV-1
HIV-2
HIV-1
Negative
HIV-1
HIV-2
HIV-1
Repeatability*
S.D.
C.V.(%)
0.007
9.9
0.071
1.2
0.046
1.1
0.023
1.8
0.006
7.6
0.169
2.5
0.060
1.4
0.045
3.2
0.003
4.4
0.085
1.4
0.033
0.7
0.028
2.1
Between Day**
S.D.
C.V.(%)
0.005
6.9
0.172
2.8
0.163
3.9
0.046
3.5
0.003
4.3
0.163
2.5
0.100
2.3
0.035
2.5
0.004
5.6
0.131
2.1
0.158
3.6
0.034
2.5
Total***
S.D.
C.V.(%)
0.008
12.1
0.186
3.1
0.169
4.1
0.052
4.0
0.007
8.7
0.235
3.5
0.117
2.7
0.057
4.1
0.005
7.1
0.156
2.5
0.162
3.7
0.044
3.3
No. of
Obs.
No. of
Days
40
40
40
40
40
40
40
40
40
40
40
40
20
20
20
20
20
20
20
20
20
20
20
20
* Repeatability: Variability of the assay performance from replicate to replicate.
** Between Day: Variability of the assay performance from day to day.
*** Total: Variability of the assay combining the effects of repeatability and between day.
Within occasion
CV = 1.9%
Day to day
CV = 2.9%
Total
CV = 3.5%
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Result Algorithm
An initial singleton
result of <0.90 s/c
indicates a nonreactive Sample
that is “Negative”
for Anti-HIV 1+2
An initial result of >/= 0.90 and <
1.00 s/c (“Retest?”) indicates a
sample that requires duplicate
repeat testing for
Anti-HIV 1+2
Retest in
Duplicate
If 2 of 3 results < 1.00 S/C,
the Sample is “Negative” for
Anti- HIV 1+2
No further testing
required. “Negative”
An initial singleton
result of >/= 1.00 s/c
indicates a sample
that is “Reactive”
for Anti-HIV 1+2
If 2 of 3 results >/= 1.00 S/C,
the Sample is “Reactive” for
Anti- HIV 1+2
No repeat testing required:
Perform Supplemental
Testing as Appropriate 13
Mistake proofing diagnostic systems
– Integrated Process Control
• Series of technologies that perform, monitor, verify
and document diagnostic checks throughout sample
and assay processing and reporting results
• Sample and Result Integrity Verification
– Traceability with real-time operator notifications and
documentation for exceptions
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Intellicheck Example: Sample Integrity Verification
Patented Pressure Level-Sensing Technology
Bubble, Clot, Viscosity, Short Sample, Thin Layer Fluid
detection
Positive pressure
detects
Negative pressure
aspirates
Normal
Aspiration
Abnormal
Aspirate bubble
detected
Pressure profile detects anomalies
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Summary
Parameter
Clinical
Performance
Performance
Positive Percent Agreement
99.94% (1653/1654)
Negative Percent Agreement 99.70% (4252/4265)
Excellent separation of positive and negative populations
Precision
Total precision of 3.5% in PMA studies
Genotypes
All genotypes detected
Seroconversion
Early detection of seroconversions
Sample type
Validated for serum and plasma with normal, high risk and
pediatric populations.
Resolution
Algorithm
Singleton testing for both negative and positive samples.
Repeat testing only for borderline samples.
Calibration and
controls
Calibration only required once every 28 days
Controls run once every 24 hours of testing
System
Control of a random access system with the security of
Intellicheck ®
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