A Pragmatic Approach to HIV-1 Drug Resistance Determination in Resource-Limited Settings by Use of a Novel Genotyping Assay Targeting the Reverse Transcriptase-Encoding Region Only

Aitken, Sue; Bronze, Michelle; Wallis, Carole L.; Stuyver, Lieven; Steegen, Kim; Balinda, Sheila N.; Kityo, Cissy; Stevens, Wendy; Wit, Tobias F. Rinke de; Schuurman, Rob

doi:10.1128/jcm.00118-13

Cited by 21 publications

(14 citation statements)

References 13 publications

(21 reference statements)

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“…Ideally, this may change in the future due to efforts to develop simplified tests against lower costs, facilitating individualized testing in resource-limited settings [36]. However, until then, drug resistance surveillance programmes on a national level could provide valuable information on PDR prevalence [37]. This is especially important in children given the high prevalence of paediatric PDR and the relation with treatment failure, and because second-line drugs options are limited in this population.…”

Section: Discussionmentioning

confidence: 99%

High levels of pre‐treatment HIV drug resistance and treatment failure in Nigerian children

Boerma

Boender

Sigaloff

et al. 2016

Journal of the International AIDS Society

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IntroductionPre-treatment HIV drug resistance (PDR) is an increasing problem in sub-Saharan Africa. Children are an especially vulnerable population to develop PDR given that paediatric second-line treatment options are limited. Although monitoring of PDR is important, data on the paediatric prevalence in sub-Saharan Africa and its consequences for treatment outcomes are scarce. We designed a prospective paediatric cohort study to document the prevalence of PDR and its effect on subsequent treatment failure in Nigeria, the country with the second highest number of HIV-infected children in the world.MethodsHIV-1-infected children ≤12 years, who had not been exposed to drugs for the prevention of mother-to-child transmission (PMTCT), were enrolled between 2012 and 2013, and followed up for 24 months in Lagos, Nigeria. Pre-antiretroviral treatment (ART) population-based pol genotypic testing and six-monthly viral load (VL) testing were performed. Logistic regression analysis was used to assess the effect of PDR (World Health Organization (WHO) list for transmitted drug resistance) on subsequent treatment failure (two consecutive VL measurements >1000 cps/ml or death).ResultsOf the total 82 PMTCT-naïve children, 13 (15.9%) had PDR. All 13 children harboured non-nucleoside reverse transcriptase inhibitor (NNRTI) mutations, of whom seven also had nucleoside reverse transcriptase inhibitor resistance. After 24 months, 33% had experienced treatment failure. Treatment failure was associated with PDR and a higher log VL before treatment initiation (adjusted odds ratio (aOR) 7.53 (95%CI 1.61–35.15) and 2.85 (95%CI 1.04–7.78), respectively).DiscussionPDR was present in one out of six Nigerian children. These high numbers corroborate with recent findings in other African countries. The presence of PDR was relevant as it was the strongest predictor of first-line treatment failure.ConclusionsOur findings stress the importance of implementing fully active regimens in children living with HIV. This includes the implementation of protease inhibitor (PI)-based first-line ART, as is recommended by the WHO for all HIV-infected children <3 years of age. Overcoming practical barriers to implement PI-based regimens is essential to ensure optimal treatment for HIV-infected children in sub-Saharan Africa. In countries where individual VL or resistance testing is not possible, more attention should be given to paediatric PDR surveys.

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Section: Discussionmentioning

confidence: 99%

High levels of pre‐treatment HIV drug resistance and treatment failure in Nigerian children

Boerma

Boender

Sigaloff

et al. 2016

Journal of the International AIDS Society

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“…Isolated NA were amplified and genotyped using an assay targeting a region of the reverse transcriptase (RT) gene from amino acid 41–238 [ 30 ], which has a level of detection of log 10 3.7 RNA copies/ml. Briefly, the assay amplifies a 560 nucleotide fragment in a single RT-PCR, which is subsequently sequenced using a single forward and a single reverse primer.…”

Section: Methodsmentioning

confidence: 99%

Stability of HIV-1 Nucleic Acids in Dried Blood Spot Samples for HIV-1 Drug Resistance Genotyping

et al. 2015

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Dried blood spots (DBS) are an easy to collect sample-type that can stabilize biological material at ambient temperature for transport and storage, making them ideal for use in resource-limited settings (RLS). We investigated the effect of storage temperature and duration on ability to detect mixed HIV-1 viral RNA populations, and subsequently viral RNA populations in a background of proviral DNA. Part one of the study used DBS samples of whole blood spiked with specific quantities of HIV-1 subtype-B and -C RNA to study mixed virus population detection. Part two used DBS comprising of HIV-1 subtype-B proviral DNA containing U1 cells combined with HIV-1 subtype-C RNA to mimic HIV-1 infected clinical samples as a model system to study the relative stability of HIV-1 RNA and DNA in DBS. Prepared DBS were stored at -20°C and +30°C for periods of one day, one, two, and four weeks. Samples were genotyped to determine changes in the detection of mixtures in the sample over time. From two weeks onwards, storage at +30°C resulted in gradual, time-related reduction in the detection of mixed virus population at log10 VL 4.0 but not at log10 5.0. Proviral DNA and viral RNA were both stable for at least 52 weeks when stored at -20°C, compared to progressive RNA decay over time at +30°C. DBS storage conditions and duration had a significant effect on HIV-1 RNA amplification. Our results demonstrate that DBS storage at ambient temperature (+30°C) should not exceed two weeks, with long-term storage at -20°C or lower.

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“…Qualitative rather than quantitative detection of failure has previously been shown to reduce the cost of virological monitoring [24], and sequencing of the reverse transcriptase coding region of the pol gene alone likewise reduces costs compared to longer sequences [25].…”

Section: Discussionmentioning

confidence: 99%

A qualitative PCR minipool strategy to screen for virologic failure and antiretroviral drug resistance in South African patients on first-line antiretroviral therapy

Newman

Breunig

Zyl

et al. 2014

Journal of Clinical Virology

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A Pragmatic Approach to HIV-1 Drug Resistance Determination in Resource-Limited Settings by Use of a Novel Genotyping Assay Targeting the Reverse Transcriptase-Encoding Region Only

Cited by 21 publications

References 13 publications

High levels of pre‐treatment HIV drug resistance and treatment failure in Nigerian children

High levels of pre‐treatment HIV drug resistance and treatment failure in Nigerian children

Stability of HIV-1 Nucleic Acids in Dried Blood Spot Samples for HIV-1 Drug Resistance Genotyping

A qualitative PCR minipool strategy to screen for virologic failure and antiretroviral drug resistance in South African patients on first-line antiretroviral therapy

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