1. Phar. Nhat Mang/ Roche Vietnam
EVALUATION THE EFFECTIVENESS OF TESTING VIRAL LOAD COBAS® AmpliPrep / COBAS® TaqMan® HIV-1 Test v2.0
Phar. Nhat Mang/ Roche Vietnam

2. Outline Background Objectives Methods Results & Discussions
Conclusions
Recommendations

3. Outline Background Objectives Methods Results & Discussions
Conclusions
Recommendations

4. <50% of patients on treatment have access to viral load testing
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
UNAIDS reports

5. Better treatment monitoring
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
Guidelines updated in July 2013, now promote the use of VL testing instead of CD4.
Also recommend starting treatment earlier and putting all HIV+ pregnant women on treatment for life
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

6. 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, non-O (N)
Subtype D
Subtype F
Subtype H
Subtype J
Subtype K
CRF_AE
Subtype G
Subtype C
Subtype B
Subtype A
Group N
Group O
CRF_AG
other CRFs
HIV has an extremely high mutation rate which differs from region to region
Roche’s Global Surveillance Program

7. 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
Ensures enhanced reliability of test results and more confidence in assessing viral loads

8. 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 ?

9. Outline Background Objectives Methods Results & Discussions
Conclusions
Recommendations

10. 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
Mutations can lead to under-quantitation or non-detection of patient specimen

11. Outline Background Objectives Methods Results & Discussions
Conclusions
Recommendations

12. 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

13. 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
A dual target builds redundancy into the assay; potentially avoids underquantification
Cobas HIV-1 v2.0 chose gag & LTR region as assay’s target
LOD : 20 copies/ml
Arellano, E. Virus Genes :111-6

14. 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
A dual target builds redundancy into the assay; potentially avoids underquantification
Figure: Young T P et al. J. Clin. Microbiol. 2011;49:
Abbott Package Insert
Reigadas et al. J Antimicrob Chemother Apr;68(4):969-72
Simon et al. Lancet Aug 5;368(9534):
Real-Time HIV-1 choice the INT (integrase) region (pol gene) as the assay’s target
LOD : 40 copies /ml
Ceccherini-Silberstein F et al. Characterization and structural analysis of HIV-1 integrase conservation. AIDS Rev Jan-Mar; 11: Latallaide M et al. Natural polymorphism of the HIV-1 integrase gene and mutations associated with integrase inhibitor resistance. Antivir Ther. 2007; 12:
Reigadas et al. J Antimicrob Chemother Apr;68(4):969-72
Simon et al. Lancet Aug 5;368(9534):

15. Outline Background Objectives Methods Results & Discussions
Conclusions
Recommendations

16. 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 Mar;56(3): PubMed PMID:

17. 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 Mar;56(3): PubMed PMID:

18. Comparative RNA Quantification of HIV-1
CAP/CTM HIV-1 v2.0 and Real-Time HIV-1 PCR Assays
84 samples below LOD of Abbott technique
(<40 copies/ml)
55 samples below LOD of CAP/CTM v2.0
(<20 copies/ml)
12 samples between copies/ml of CAP/CTM v2.0
17 samples above 40 copies/ml of CAP/CTM v2.0 ( range copies/ml)
A difference of individual result for two techniques
Negative by Abbott
Negative by CAP/CTM v2.0
Positive by CAP/CTM v2.0
Sire JM, et al. J Acquir Immune Defic Syndr Mar;56(3): PubMed PMID:

19. Integrase region of the pol gene*
Roche 2013
4/27/2017
What impact does this have on patients? HIV resistance in a patient likely failing treatment
Bryan – this is real for patients
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 Mar;56(3): PubMed PMID:

20. More Literature demonstrates…underquantitation!
The Abbott assay appeared to underestimate viral load 40-50% of the time compared to 2nd WHO Standard
Janse van Rensburg
30 samples not quantified by the Abbott RealTime HIV-1 assay were quantitated by COBAS® AmpliPrep/COBAS TaqMan® HIV-1 Test, v2.0
Paba
Three samples (1%) underquantified by Abbott assay by more than 1 log10 relative to COBAS® AmpliPrep/COBAS TaqMan® HIV-1 Test, v2.0
Bryan
Pas
Abbott underquants 10% of the samples (n=50, >1 log10 (cp/mL) = 5)
Church

21. Outline Background Objectives Methods Results & Discussions
Conclusions
Recommendations

22. 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
Ensures enhanced reliability of test results and more confidence in assessing viral loads

23. Outline Background Objectives Methods Results & Discussions
Conclusions
Recommendations

24. 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

25. Doing now what patients need next
Thank you
Doing now what patients need next