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THE 6TH
NATIONAL SCIENTIFIC CONFERENCE ON HIV/AIDS
EVALUATION THE EFFECTIVENESS OF TESTING VIRAL LOAD
COBAS® AmpliPrep / COBAS® TaqMan® HIV-1 Test v2.0
Phar. Nhat Mang/ Roche Vietnam
Outline
1. Background
2. Objectives
3. Methods
4. Results & Discussions
5. Conclusions
6. Recommendations
The 6th National Scientific Conference on HIV/AIDS
Outline
1. Background
2. Objectives
3. Methods
4. Results & Discussions
5. Conclusions
6. Recommendations
The 6th National Scientific Conference on HIV/AIDS
Current and goal of HIV epidemic
15 million people on treatment as of March 2015
Only 45% of
people living with
HIV know they are
infected
38% of HIV
positive patients
receive treatment
<50% of patients
on treatment
have access to
viral load testing
 Goal of ending the AIDS epidemic by 2030
The 6th National Scientific Conference on HIV/AIDS
UNAIDS reports
WHO guidelines recommend use of viral load
Driving scale up in resource limited settings
Better
treatment
monitoring
• Use of Viral Load instead of
CD4
26 Million
• Number of people now
eligible for ART, and increase
of over 10 million
3 Million
• Number of deaths averted by
2025 if guidelines are
implemented
 HIV viral load assay is the important step to monitor the effectiveness of
ARV treatment and also follow up the progression of HIV disease
WHO guidelines 2013
The 6th National Scientific Conference on HIV/AIDS
Overview HIV
Monitoring HIV-1 genomes to stay state-of-the-art
 HIV is a retrovirus and belongs to the sub-family of lentiviruses.
 HIV type 1 and HIV type 2 can both cause AIDS although the latter
causes a milder and slower disease.
 HIV-1 is divided into three main groups: Main (M), Outilier (O) and non-M,
CRF_AG
non-O (N)
CRF_AE
Group O
other CRFs
Group N
Subtype K
Subtype A
Subtype J
Subtype H
Subtype G
Subtype F
Subtype D
Subtype B
Subtype C
Roche’s Global Surveillance Program
The 6th National Scientific Conference on HIV/AIDS
The technique of viral load testing
3 techniques for
 Real time PCR
 bDNA (branched-chain DNA)
 NASBA (nucleic acid sequence based amplification)
 Real time PCR is the most common method
achievement the main goals of accuracy/reliability of test
results
 Sensitivity
 Specificity
 Wide detection range
The 6th National Scientific Conference on HIV/AIDS
The technique of viral load testing
Real time PCR method focus
 Single target technology
 Dual target technology
DUAL
TARGET
Target 1
SINGLE
TARGET
DUAL
TARGET
Target 2
 What is the better technology and most reliable in assessing viral
loads ?
The 6th National Scientific Conference on HIV/AIDS
Outline
1. Background
2. Objectives
3. Methods
4. Results & Discussions
5. Conclusions
6. Recommendations
The 6th National Scientific Conference on HIV/AIDS
Comparative RNA Quantification of HIV-1
CAP/CTM HIV-1 v2.0 and Real-Time HIV-1 PCR Assays
 Objective 1: To compare the effectiveness of detection
of Roche CAP/CTM HIV-1 v2.0 and Real-Time HIV-1
PCR Assays for the detection and quantification of HIV-1
RNA in plasma sample containing different HIV-1 groups
and subtypes
 Objective 2: Reinforce the DUAL target technology to
enhances reliability and minimizes the effects against
natural mutations than SINGLE Target
The 6th National Scientific Conference on HIV/AIDS
Outline
1. Background
2. Objectives
3. Methods
4. Results & Discussions
5. Conclusions
6. Recommendations
The 6th National Scientific Conference on HIV/AIDS
Viral load measurements for patients
CAP/CTM HIV-1 v2.0 and Real-Time HIV-1 PCR Assays
These PCR assays were implemented with
 CAP/CTM HIV-1 v2.0: COBAS AmpliPrep and COBAS TaqMan
 Real-Time HIV-1
: m2000sp/m2000rt
 CAP/CTM is a fully automated system, reduces vastly the risk of
contamination, is faster and has high-throughput
The 6th National Scientific Conference on HIV/AIDS
Two (dual) targets:
COBAS® AmpliPrep / COBAS® TaqMan® HIV-1 Test v2.0
Gag:
 Targeted in previous Roche HIV-1 viral
load tests to drive correlation
 Large sequence database from Roche’s
Global surveillance program
 Not a drug target
LTR:
 Targeted in Roche MPx blood screening test
 Not a drug target
 Cobas HIV-1 v2.0 chose gag & LTR region as assay’s target
 LOD : 20 copies/ml
Arellano, E. Virus Genes 2007 34:111-6
The 6th National Scientific Conference on HIV/AIDS
Single target:
Real-Time HIV -1
int :
 Targeted in Abbott HIV-1 viral load tests
 A drug target and over 42 mutations associated
with drug resistance to Integrase Inhibitors
 Real-Time HIV-1 choice the INT (integrase) region (pol gene) as the
assay’s target
Ceccherini-Silberstein F et al. Characterization and structural analysis of HIV-1 integrase conservation.
 LOD : 40 copies /ml
AIDS Rev. 2009 Jan-Mar; 11: 17-29.
Latallaide M et al. Natural polymorphism of the HIV-1 integrase gene and mutations associated with
integrase inhibitor resistance. Antivir Ther. 2007; 12: 563-570.
Reigadas et al. J Antimicrob Chemother. 2013 Apr;68(4):969-72
Simon et al. Lancet. 2006 Aug 5;368(9534):489-504
The 6th National Scientific Conference on HIV/AIDS
Outline
1. Background
2. Objectives
3. Methods
4. Results & Discussions
5. Conclusions
6. Recommendations
The 6th National Scientific Conference on HIV/AIDS
Comparative RNA Quantification of HIV-1
CAP/CTM HIV-1 v2.0 and Real-Time HIV-1 PCR Assays
260 plasma sample from HIV-1 group M
 Roche the median pVL value : 3.08 log10 copies/ml
 Abbott the median pVL value : 2.81 log10 copies/ml
 Confirmed that the higher value of the CAP/CTM v2.0 assay
Sire JM, et al. J Acquir Immune Defic Syndr. 2011 Mar;56(3):239-43. PubMed PMID: 21164353.
The 6th National Scientific Conference on HIV/AIDS
Comparative RNA Quantification of HIV-1
CAP/CTM HIV-1 v2.0 and Real-Time HIV-1 PCR Assays
A difference greater than 0.5 log10 copies/ml is considered clinically relevant
 20% significantly lower by Abbott than COBAS® AmpliPre/COBAS
TaqMan® HIV-1 Test, v2.0
Sire JM, et al. J Acquir Immune Defic Syndr. 2011 Mar;56(3):239-43. PubMed PMID: 21164353.
The 6th National Scientific Conference on HIV/AIDS
Comparative RNA Quantification of HIV-1
CAP/CTM HIV-1 v2.0 and Real-Time HIV-1 PCR Assays
A difference of individual result for two techniques
84 samples below
LOD of Abbott
technique
(<40 copies/ml)
55 samples below
LOD of CAP/CTM v2.0
(<20 copies/ml)
Negative by
CAP/CTM v2.0
12 samples between
20-40 copies/ml of
CAP/CTM v2.0
Negative by
Abbott
17 samples above 40
copies/ml of CAP/CTM
v2.0 ( range 41-897
copies/ml)
Positive by CAP/CTM v2.0
Sire JM, et al. J Acquir Immune Defic Syndr. 2011 Mar;56(3):239-43. PubMed PMID: 21164353.
The 6th National Scientific Conference on HIV/AIDS
What impact does this have on patients?
HIV resistance in a patient likely failing treatment
Integrase region of the pol gene*
 HIV resistance in a patient likely failing treatment may not be detected with the Abbott HIV Test
Sire JM, et al. J Acquir Immune Defic Syndr. 2011 Mar;56(3):239-43. PubMed PMID: 21164353.
The 6th National Scientific Conference on HIV/AIDS
More Literature
demonstrates…underquantitation!
Janse van
Rensburg
Paba
Pas
Church
The Abbott assay appeared to
underestimate viral load 40-50% of the time
compared to 2nd WHO Standard
30 samples not quantified by the Abbott
RealTime HIV-1 assay were quantitated by
COBAS® AmpliPrep/COBAS TaqMan®
HIV-1 Test, v2.0
Three samples (1%) underquantified by
Abbott assay by more than 1 log10
relative to COBAS® AmpliPrep/COBAS
TaqMan® HIV-1 Test, v2.0
Abbott underquants 10% of the
samples (n=50, >1 log10 (cp/mL) = 5)
The 6th National Scientific Conference on HIV/AIDS
Outline
1. Background
2. Objectives
3. Methods
4. Results & Discussions
5. Conclusions
6. Recommendations
The 6th National Scientific Conference on HIV/AIDS
Comparative RNA Quantification of HIV-1
Stay One Step Ahead with the HIV-1 dual target assay
 Objective 1:

Higher values obtained with CAP/CTM test v2.0 test relative to the Abbott test

Higher sensitivity of CAP/CTM test v2.0 assay (20 copies) compared with the Abbott
test

The higher meaningful for clinicians and patient because the CAP/CTM test v2.0 test
may show low-level viral load in patients whose viral load is undetectable with other
methods
 Objective 2:

Abbott – Single target might be impaired by mutations in the integrase target region.
Beside that Roche CAP/CTM dual target builds redundancy into the assay,
potentially avoids underquantification
The 6th National Scientific Conference on HIV/AIDS
Outline
1. Background
2. Objectives
3. Methods
4. Results & Discussions
5. Conclusions
6. Recommendations
The 6th National Scientific Conference on HIV/AIDS
Value of result ……
COBAS® AmpliPrep / COBAS® TaqMan® HIV-1 Test v2.0
 Provides diagnostic accuracy of test results even if mutations occur
in one of the two regions
 Dual target minimizes the effects against MUTATIONS and also
represents an important step forward
 Is fully traceable to WHO international standards and CE-IVD
certificated
The 6th National Scientific Conference on HIV/AIDS
Thank you
Doing now what patients need next
The 6th National Scientific Conference on HIV/AIDS