Highlights
- •DBS are useful for HIV viraemia testing in remote settings.
- •Viral load testing can be affected by HIV genetic variability and by assay election.
- •HIV-1 quantification values can be different using distinct assays.
- •Viral load assays should increase their sensitivity and specificity.
- •We recommend using the same VL technique for each patient during ART monitoring.
Summary
Objectives
Viral load (VL) testing is used for early HIV diagnosis in infants (EID) and for detecting
early therapeutic failure events, but can be affected by HIV genetic variability.
Dried blood samples (DBS) increase VL access and EID in remote settings and when low
blood volume is available.
Methods
This study compares VL values using Siemens VERSANT HIV-1 RNA 1.0 kPCR assay (kPCR)
and Roche CAP/CTM Quantitative test v2.0 (CAP/CTM v2.0) in 176 DBS carrying different
HIV-1 variants collected from 69 Equatoguinean mothers and their infants with known
HIV-1 status (71 infected, 105 uninfected).
Results
CAP/CTM v2.0 provided false positive VLs in 11 (10.5%) cases. VL differences above
0.5 log10 were observed in 42/49 (87.5%) DBS, and were above 1 log10 in 18 cases. CAP/CTM v2.0 quantified all the 41 specimens with previously inferred
HIV-1 variant by phylogenetic analysis (68.3% recombinants) whereas kPCR only identified
90.2% of them, and was unable to detect 14.3% of 21 CRF02_AG viruses. CAP/CTM v2.0
showed higher sensitivity than kPCR (95.8% vs. 70.1%), quantifying a higher rate of viruses in infected DBS from subjects under
antiretroviral exposure at sampling time compared to kPCR (94.7% vs. 96.2%, p-value<0.001).
kPCR showed maximum specificity (100%) whereas for CAP/CTM v2.0 was 89.5%.
Conclusions
VL assays should increase their sensitivity and specificity to avoid overestimated
HIV-1 quantifications, which could be interpreted as virological failure events, or
false negative diagnostic results due to genetic variability. We recommend using the
same VL technique for each patient during antiretroviral therapy monitoring.
Keywords
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References
- In-house HIV-1 RNA real-time RT-PCR assays: principle, available tests and usefulness in developing countries.Expert Rev Mol Diagn. 2008; 8: 635-650https://doi.org/10.1586/14737159.8.5.635
- Successes, challenges, and limitations of current antiretroviral therapy in low-income and middle-income countries.Lancet Infect Dis. 2009; 9: 637-649https://doi.org/10.1016/S1473-3099(09)70227-0
- Dried blood spots can expand access to virological monitoring of HIV treatment in resource-limited settings.J Antimicrob Chemother. 2009; 64: 1126-1129https://doi.org/10.1093/jac/dkp353
- Dried blood spot HIV-1 RNA quantification using open real-time systems in South Africa and Burkina Faso.J Acquir Immune Defic Syndr. 2010; 55: 290-298https://doi.org/10.1097/QAI.0b013e3181edaaf5
- Outcomes of antiretroviral treatment in programmes with and without routine viral load monitoring in Southern Africa.AIDS. 2011; 25: 1761-1769https://doi.org/10.1097/QAD.0b013e328349822f
- HIV-1 viral load monitoring: an opportunity to reinforce treatment adherence in a resource-limited setting in Thailand.Trans R Soc Trop Med Hyg. 2009; 103: 601-606https://doi.org/10.1016/j.trstmh.2008.11.007
- Viral load monitoring as a tool to reinforce adherence: a systematic review.J Acquir Immune Defic Syndr. 2013; 64: 74-78https://doi.org/10.1097/QAI.0b013e31829f05ac
- Technical and operational considerations for implementing HIV viral load testing.2014 (Available:) ([accessed 19.02.15])
- Challenges and opportunities for the implementation of virological testing in resource-limited settings.J Int AIDS Soc. 2012; 15: 17324https://doi.org/10.7448/IAS.15.2.17324
- Expanding access to HIV viral load testing: a systematic review of RNA stability in EDTA tubes and PPT beyond current time and temperature thresholds.PLoS One. 2014; 9: e113813https://doi.org/10.1371/journal.pone.0113813
- Multicenter evaluation of use of dried blood and plasma spot specimens in quantitative assays for human immunodeficiency virus RNA: measurement, precision, and RNA stability.J Clin Microbiol. 2003; 41: 1888-1893
- Dried blood spots for the diagnosis and quantitation of HIV-1: stability studies and evaluation of sensitivity and specificity for the diagnosis of infant HIV-1 infection in Thailand.J Virol Methods. 2009; 155: 109-117https://doi.org/10.1016/j.jviromet.2008.09.022
- Dried blood spots for monitoring HIV infection in Public Health Programs in developing countries.Enferm Infecc Microbiol Clin. 2013; 31: 100-107https://doi.org/10.1016/j.eimc.2012.03.008
- Dried blood spots improve access to HIV diagnosis and care for infants in low-resource settings.J Acquir Immune Defic Syndr. 2005; 38: 615-617
- Systematic review of the use of dried blood spots for monitoring HIV viral load and for early infant diagnosis.PLoS One. 2014; 9: e86461https://doi.org/10.1371/journal.pone.0086461
- Evaluation of a dried blood spot HIV-1 RNA program for early infant diagnosis and viral load monitoring at rural and remote healthcare facilities.AIDS. 2009; 23: 2459-2466https://doi.org/10.1097/QAD.0b013e328331f702
- Implementing early infant diagnosis of HIV infection at the primary care level: experiences and challenges in Malawi.Bull World Health Organ. 2012; 90: 699-704
- Evaluation of dried blood spot diagnosis using HIV1-DNA and HIV1-RNA Biocentric assays in infants in Abidjan, Côte d'Ivoire. The Pedi-Test DBS ANRS 12183 Study.J Virol Methods. 2013; 193: 439-445https://doi.org/10.1016/j.jviromet.2013.07.011
- Limited utility of dried-blood- and plasma spot-based screening for antiretroviral treatment failure with Cobas Ampliprep/TaqMan HIV-1 version 2.0.J Clin Microbiol. 2014; 52: 3878-3883https://doi.org/10.1128/JCM.02063-14
- Evaluation of filter paper transfer of whole-blood and plasma samples for quantifying HIV RNA in subjects on antiretroviral therapy in Uganda.J Acquir Immune Defic Syndr. 2007; 46: 590-593
- Correlation between HIV-1 viral load quantification in plasma, dried blood spots, and dried plasma spots using the Roche COBAS Taqman assay.J Clin Virol. 2010; 47: 4-7https://doi.org/10.1016/j.jcv.2009.11.006
- HIV-1 RNA quantification from Dried Blood Spots and plasma using the Siemens VERSANT® HIV-1 RNA 1.0 Assay (kPCR).J Virol Antivir Res. 2014; 3: 2https://doi.org/10.4172/2324-8955.1000123
- Performance of three commercial viral load assays, Versant human immunodeficiency virus type 1 (HIV-1) RNA bDNA v3.0, Cobas AmpliPrep/Cobas TaqMan HIV-1, and NucliSens HIV-1 EasyQ v1.2, testing HIV-1 non-B subtypes and recombinant variants.J Clin Microbiol. 2008; 46: 2918-2923https://doi.org/10.1128/JCM.02414-07
- HIV-1 viral diversity and its implications for viral load testing: review of current platforms.Int J Infect Dis. 2011; 15: e661-670https://doi.org/10.1016/j.ijid.2011.05.013
- Current challenges to viral load testing in the context of emerging genetic diversity of HIV-1.Expert Opin Med Diagn. 2011; 5: 183-202https://doi.org/10.1517/17530059.2011.566860
- The first B/G intersubtype recombinant form of human immunodeficiency virus type 1 (HIV-1) identified in Germany was undetected or underquantitated by some commercial viral load assays.J Med Virol. 2006; 78: 311-317
- Significant impact of non-B HIV-1 variants genetic diversity in Gabon on plasma HIV-1 RNA quantitation.J Med Virol. 2014; 86: 52-57https://doi.org/10.1002/jmv.23770
- Discordances with HIV-1 RNA quantitative determinations by three commercial assays in Pointe Noire, Republic of Congo.J Virol Methods. 2014; 203: 102https://doi.org/10.1016/j.jviromet.2014.02.030
- Equatorial Guinea National Progress Report.2014 (Available:) ([accessed 19.02.15])
- Description of HIV-1 group M molecular epidemiology and drug resistance prevalence in Equatorial Guinea from migrants in Spain.PLoS One. 2013; 8: e64293https://doi.org/10.1371/journal.pone.0064293
- Molecular epidemiology of HIV type 1 subtypes in Equatorial Guinea.AIDS Res Hum Retroviruses. 2001; 17: 851-855
- The prevalence of diverse HIV-1 strains was stable in Cameroonian blood donors from 1996 to 2004.J Acquir Immune Defic Syndr. 2008; 49: 432-439https://doi.org/10.1097/QAI.0b013e31818a6561
- Global trends in molecular epidemiology of HIV-1 during 2000-2007.AIDS. 2011; 25: 679-689https://doi.org/10.1097/QAD.0b013e328342ff93
- Selected hematologic and biochemical measurements in African HIV-infected and uninfected pregnant women and their infants: the HIV Prevention Trials Network 024 protocol.BMC Pediatr. 2009; 9: 49https://doi.org/10.1186/1471-2431-9-49
- Recommendations on the diagnosis of HIV infection in infants and children.2010 (Available at:) ([accessed 19.02.15])
- The use of antiretroviral drugs for treating and preventing HIV infection.2013 (Available at:) ([accessed 19.02.15])
- March 2014 supplement to the 2013 consolidated guidelines on the use of antiretroviral drugs for treating and preventing HIV infection.2014 (Available at:) ([accessed 19.02.15])
- Protocolo de diagnóstico y seguimiento por laboratorio de VIH/SIDA en Guinea Ecuatorial.1st ed. MINSABS, Malabo2009
- HIV-1 infection can be confirmed by Bio-Rad GeeniusTM HIV 1/2 confirmatory assay using dried blood spots.J Clin Virol. 2015; 63: 66-69https://doi.org/10.1016/j.jcv.2014.12.018
- Performance evaluation of the new Roche Cobas AmpliPrep/Cobas TaqMan HIV-1 test version 2.0 for quantification of human immunodeficiency virus type 1 RNA.J Clin Microbiol. 2010; 48: 1195-1200https://doi.org/10.1128/JCM.01832-09
- Evaluation of the Roche Cobas AmpliPrep/Cobas TaqMan HIV-1 test and identification of rare polymorphisms potentially affecting assay performance.J Clin Microbiol. 2010; 48: 2852-2858https://doi.org/10.1128/JCM.00776-10
- Evaluation of the NucliSens EasyQ v2.0 assay in comparison with the Roche Amplicor v1.5 and the Roche CAP/CTM HIV-1 Test v2.0 in quantification of C-clade HIV-1 in plasma.PLoS One. 2014; 9: e103983https://doi.org/10.1371/journal.pone.0103983
- Establishing diagnostic cut-off criteria for the COBAS AmpliPrep/COBAS TaqMan HIV-1 Qualitative test through validation against the Amplicor DNA test v1.5 for infant diagnosis using dried blood spots.J Clin Virol. 2012; 53: 106-109https://doi.org/10.1016/j.jcv.2011.12.002
Troppan KT, Stelzl E, Violan D, Winkler M, Kessler HH. Evaluation of the new VERSANT HIV-1 RNA 1.0 Assay (kPCR) for quantitative detection of human immunodeficiency virus type 1 RNA. J Clin Virol. 46:69–74. doi: 10.1016/j.jcv.2009.06.012.
- Quantification of HIV-RNA from dried blood spots using the Siemens VERSANT® HIV-1 RNA (kPCR) assay.J Antimicrob Chemother. 2011; 66: 2823-2826https://doi.org/10.1093/jac/dkr383
- Analytical performance of an automated assay quantifying HIV-1 from dried blood spots.J Clin Virol. 2013; 57: 271-273https://doi.org/10.1016/j.jcv.2013.03.001
- Comparing methods of measurement: why plotting difference against standard method is misleading.Lancet. 1995; 346: 1085-1087
- Monitoring for treatment failure in patients on first-line antiretroviral treatment in resource-constrained settings.Curr Opin HIV AIDS. 2010; 5: 1-5https://doi.org/10.1097/COH.0b013e3283333762
- Antiretroviral therapy for HIV infection in adults and adolescents: recommendations for a public health approach.WHO Press, Geneva, Switzerland2010 (Available:) ([accessed 19.02.15])
- Unnecessary antiretroviral treatment switches and accumulation of HIV resistance mutations; two arguments for viral load monitoring in Africa.J Acquir Immune Defic Syndr. 2011; 58: 23-31https://doi.org/10.1097/QAI.0b013e318227fc34
- Accumulation of drug resistance and loss of therapeutic options precede commonly used criteria for treatment failure in HIV-1 subtype-C-infected patients.Antivir Ther. 2012; 17: 377-386https://doi.org/10.3851/IMP2010
- Evaluation of quantification of HIV-1 RNA viral load in plasma and dried blood spots by use of the semiautomated Cobas Amplicor assay and the fully automated Cobas Ampliprep/TaqMan assay, version 2.0, in Kisumu.Kenya.J Clin Microbiol. 2013; 51: 1208-1218https://doi.org/10.1128/JCM.03048-12
- Quantitation of human immunodeficiency virus type 1 RNA in plasma by using blood dried on filter paper.J Clin Microbiol. 1998; 36: 258-260
- Evaluation of different RNA extraction methods and storage conditions of dried plasma or blood spots for human immunodeficiency virus type 1 RNA quantification and PCR amplification for drug resistance testing.J Clin Microbiol. 2009; 47: 1107-1118https://doi.org/10.1128/JCM.02255-08
- HIV load testing with small samples of whole blood.J Clin Microbiol. 2010; 48: 2786-2792https://doi.org/10.1128/JCM.02276-09
- The significance of low-level plasma HIV viral load on COBAS TaqMan HIV-1 assays for patients with undetectable plasma viral load on COBAS Amplicor monitor version 1.5.HIV Clin Trials. 2008; 9: 283-286https://doi.org/10.1310/hct0904-283
- Comparative frequencies of HIV low-level viremia between real-time viral load assays at clinically relevant thresholds.J Clin Virol. 2011; 52: S83-S89https://doi.org/10.1016/j.jcv.2011.09.022
- Increased reporting of detectable plasma HIV-1 RNA levels at the critical threshold of 50 copies per milliliter with the Taqman assay in comparison to the Amplicor assay.J Acquir Immune Defic Syndr. 2009; 51: 3-6https://doi.org/10.1097/QAI.0b013e31819e721b
- Comparative evaluation of the VERSANT HIV-1 RNA 1.0 kinetic PCR molecular system (kPCR) for the quantification of HIV-1 plasma viral load.J Clin Virol. 2009; 44: 297-301https://doi.org/10.1016/j.jcv.2009.01.004
- Genotype impact on HCV RNA levels determined with the VERSANT HCV RNA 1.0 assay (kPCR).J Clin Virol. 2013; 58: 522-527https://doi.org/10.1016/j.jcv.2013.09.005
Article info
Publication history
Published online: May 29, 2015
Accepted:
May 25,
2015
Identification
Copyright
© 2015 The British Infection Association. Published by Elsevier Inc. All rights reserved.
