evaluation of commercially available hiv assays to address alternative screening/diagnostic...
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![Page 1: Evaluation of Commercially Available HIV Assays to Address Alternative Screening/Diagnostic Algorithms S. Michele Owen, Ph.D. Laboratory Branch Division](https://reader035.vdocument.in/reader035/viewer/2022080915/56649d8a5503460f94a70030/html5/thumbnails/1.jpg)
Evaluation of Commercially Available HIV Assays to Address Alternative Screening/Diagnostic
Algorithms
S. Michele Owen, Ph.D.
Laboratory Branch
Division of HIV AIDS Prevention
Centers for Disease Control and Prevention
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Background• Why Evaluate Tests and Consider Alternative Algorithms?
– Abundance: Multiple new FDA approved testing methods- NAT, Rapid Tests, New EIAs
– Ambiguity: reduce or eliminate WB indeterminate results
– Cost and Efficiency: sequential EIAs have been effectively used internationally (WHO/UNAIDS)
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Objective
Compare performance of commercially available HIV tests as a basis for evaluating alternative algorithms for HIV diagnostics or surveillance.
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Methods- Samples
• 1002 specimens from the U.S. (Boston Biomedica Inc.) Plasma centers or blood banks.
• 62 non-U.S. samples (Boston Biomedica Inc) World Wide Performance Panels
• 205 non-U.S. samples (Cameroon Blood Bank Study) Units that were reactive in one or more screening
tests for HIV, HBV, HCV or Syphilis
• All samples were collected, processed and stored using standard diagnostic protocols
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Methods-Testing
• Plasma samples: randomized and blinded tested by 6 EIAs, 4 rapid tests and 3* NAT- based tests.
• Any sample found to be reactive by any of the above tests was subjected to Western Blot
• Sample was considered to be: Positive if the Western Blot was positive (current “gold standard”) Negative if any sample was either
negative by all 13 tests described above or Western Blot negative.
• All tests were performed by trained laboratory personnel (Gen-Probe and AmpliScreen NAT testing was done by individuals certified by the company to run the test)
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EIAs EvaluatedEIA ComponentsBioMerieux Vironostika HIV-1 (2nd) HIV-1 viral lysate
BioMerieux Vironostika HIV-1 Plus O (2nd) HIV-1 viral lysate, purified viral env proteins, and synthetic peptide from transmembrane epitope of HIV-1 Group O
BIO-RAD Genetic Systems rLAV (2nd) LAV lysate and recombinant gp41
BIO-RAD Genetic Systems 1/ 2 Peptide (2nd) Synthetic peptides from env and pol regions of both HIV-1 and HIV–2
BIO-RAD Genetic Systems HIV 1/2 Plus (3rd)HIV-1 recombinant gp160 and p24, HIVgp36 synthetic peptide, and HIV-1 group O synthetic oligopeptide
Abbott HIVAB HIV-1/HIV-2 (rDNA) (3rd) Recombinant HIV-1 core and env proteins and HIV-2 env protein
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Rapid ComponentsMedMira Reveal conserved immunodominant
peptides
OraSure OraQuick peptides, gp41,gp36
Trinity Biotech Uni-Gold Recombigen recombinant immunodominant proteins
BIO-RAD Multispot HIV-2 gp36 peptide, HIV-1 gp41 peptide, recombinant gp41
Western BlotCalypte Biomedical Cambridge Biotech HIV-1 H9/HTLV-IIIB Lysate
BIO-RAD Genetic Systems HIV-1 CEM/HIV LAV Lysate
NAAT
Gen-Probe Procleix LTR and Pol
Roche Ampliscreen Gag
In house LTR
Rapid, NAAT, WB Tests
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EIA Sensitivity and SpecificityRelative to WB
Test Sensitivity Specificity
Bio-Rad Genetic Systems rLAV
n=1264
96.9 98.6
Bio-Rad Genetic Systems 1/2 Peptide
n=1264
98.1 98.6
Biomeriuex Vironostika
n=1264
98.4 96.5
Biomerieux Vironostika Plus O
n=1264
98.9 96.6
Bio-Rad Genetic Systems 1/2 Plus O
n=1264
99.4 95.8
Abbott HIV1/2 rDNA
n=1264
98.8 96.3
Sensitivity range 96.9-99.4 % Specificity range 95.8-98.6%
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Rapid Test Sensitivity and SpecificityRelative to WB
Test Sensitivity Specificity
OraSure OraQuick
n=1264
98.0 98.9
MedMira Reveal G-1
n=1264
98.4 98.4
Trinity Biotech
Uni-Gold Recombigen HIV
n=1197
98.5 99.4
BIO-RAD Multi-Spot
n=83
97.4 97.8
Sensitivity range 97.4-98.5% Specificity range 97.8-99.4%
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NAT Sensitivity and SpecificityRelative to WB
Test Sensitivity Specificity
Gen-Probe Procleix
n=1264
96.7 98.7
Roche Ampliscreen
n=1171
92.6 96.8
CDC In- House NAT
n=1264
94.9 98.8
Sensitivity range 92.6-96.7% Specificity range 96.8-98.8%
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Indeterminate Characteristics• 58 indeterminate samples
5 U.S. plasma donors 52 Cameroon samples 1 BBI non-U.S. performance panel sample
• Most had 3 or fewer EIA/Rapid positive results low S/CO values on EIA One or few bands on WB
p24 >> p66 > p55
• 4 samples positive on multiple EIAs 2/4 positive by NAT 1/4 almost complete WB pattern (known O from Spain) 3/4 p24 Ag positive (BBI)
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Current Diagnostic Algorithm Screening EIA
Non-ReactiveReactive
Repeat EIA (duplicate) Negative
+/+ +/- -/-
NegativeWB
Pos Neg Ind*
WB
Pos Neg Ind* * follow-up sample, HIV-2
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Potential Simple Algorithms
• EIA Screen /NAAT Confirmation
• EIA Screen/Rapid Confirmation
• EIA Screen/Alternate EIA Confirmation
• Rapid Screen/Alternate Rapid Confirmation
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Summary Potential Algorithms Relative to Current
EIA/WB
False Negative False Positive
EIA/NAAT 3.3% (25) 0%
EIA/Rapid 1.3% (9) 0%
EIA/EIA 0.7% (5) 0%
Rapid/Rapid 1.7% (12) 0%
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Proposed Blood Bank Algorithm HIV EIA Repeat Reactive
No WB or Alternate EIARequired(optional)
(BIO-RAD Plus)
NAT
Non-ReactiveReactive
Alternate EIA
Reactive Non-reactive
713
67538
Vironostika Plus O
Gen-Probe
1226
675 HIV-1 Positive WB
Reactive Non-Reactive
17 0
IND9
Indeterminates from 58 to 9
12 Negative2/12 False Negative
True answer for indeterminates???
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Important Caveats
• No follow-up samples available for discordant samples (true answer unknown)
• Limited demographic or epidemiological data available
• Collection, processing, and storage of samples was conducted using routine diagnostic procedures.1 freeze/thaw of specimen prior to NAT testing1-2 freeze/thaws prior to Serological testing
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Summary• Range of sensitivity observed for all tests was
92.1% - 99.4%• Range of specificity observed for all tests was
95.8% - 98.8%• Discordant results between serological and NAT-
based tests were observed.– True answer unknown
• Most indeterminate samples– Non-U.S.– Few WB Bands– Low EIA S/CO values
• 4 Indeterminate samples likely or known positive
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Conclusions• All FDA approved HIV detection assays have comparable
Sensitivity and Specificity Lower values may be due to the stringent testing methods
employed in the study
• NAAT alone can not replace WB for confirmation
• EIA/EIA, EIA/Rapid or Rapid/Rapid algorithms yielded better sensitivity than EIA combined with NAT
• Proposed Blood Bank algorithm would likely reduce indeterminate WB results
• Much work left to do to establish “best algorithm”– Seroconversion samples– Discordant/Indeterminate samples with follow-up
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AcknowledgementsIndustryBioMerieuxBIO-RADMedMiraTrinity BiotechGen-ProbeRoche
l
CDCChunfu Yang
Wei Luo
Chou Pau
Nick Delatorre
Chin-Yih OuTom SpiraBharat ParekhFaye CowartSusan KennedyDebbie KuehlDebra CandalDonna RudolphTammy BarnettSilvina MasciotraMarcia KalishSteve McDougal