Analysis of transmitted HIV-1 drug resistance using 454 ultra-deep-sequencing and the DeepChek(®)-HIV system.

Garcia-Diaz, A., McCormick, A.L., Booth, C., Gonzalez, D., Sayada, C., Haque, T., Johnson, M. and Webster, D. 2014. Analysis of transmitted HIV-1 drug resistance using 454 ultra-deep-sequencing and the DeepChek(®)-HIV system. Journal of the International AIDS Society. 17 (Suppl 3) 19752. https://doi.org/10.7448/ias.17.4.19752

TitleAnalysis of transmitted HIV-1 drug resistance using 454 ultra-deep-sequencing and the DeepChek(®)-HIV system.
TypeJournal article
AuthorsGarcia-Diaz, A., McCormick, A.L., Booth, C., Gonzalez, D., Sayada, C., Haque, T., Johnson, M. and Webster, D.
Abstract

Introduction
Next-generation sequencing (NGS) is capable of detecting resistance-associated mutations (RAMs) present at frequencies of 1% or below. Several studies have found that baseline low-frequency RAMs are associated with failure to first-line HAART. One major limitation to the expansion of this technology in routine diagnostics is the complexity and laboriousness integral to bioinformatics analysis. DeepChek (ABL, TherapyEdge) is a CE-marked software that allows automated analysis and resistance interpretation of NGS data.

Objective
To evaluate the use of 454 ultra-deep-sequencing (Roche® 454, Life Sciences; 454-UDS) and DeepChek for routine baseline resistance testing in a clinical diagnostic laboratory.

Methods
107 newly diagnosed HIV-1-infected patients (subtypes: A, n=9; B, n=52; C, n=21; D, n=2; F, n=3; G, n=1; CRF01, n=7; CRF02, n=7; CRF06, n=1; CRF07, n=1; CRF10, n=1 and unassigned complex, n=2) with a median plasma viral load of 88,727 copies/mL (range: 1380–2,143,543) were tested by 454-UDS and Sanger sequencing for the detection of protease and reverse transcriptase RAMs. In addition, integrase RAMs were investigated in 57 of them. Sequence analysis and resistance interpretation were performed using DeepChek applying 1% and 20% thresholds for variant detections; filters applied were comparison between Sanger and 454-UDS, and Stanford and IAS list for resistance interpretation.

Results
The time elapsed from generation of raw 454 data (between 2,000–5,000 sequences/sample) to elaboration of a resistance report was approximately 10 minutes per sample, equivalent to the time required for the same process using Sanger sequencing. Four patients (3.7%) showed baseline resistance by Sanger and 454-UDS at frequencies above 20%, which affected both NRTIs (n=2) and NNRTIs (n=2). In addition, 12 patients (11.2%) showed transmitted drug resistance (TDR) by 454-UDS at frequencies below 20% affecting NRTIs (n=9), NNRTIs (n=7) and PIs (n=2). Integrase resistance was not detected at baseline by 454-UDS or Sanger sequencing.

Conclusions
DeepChek allowed easy and rapid analysis and interpretation of NGS data, thus facilitating the incorporation of this technology in routine diagnostics. The use of NGS considerably increased the detection rates of TDR to NRTI, NNRTIs and PIs. No transmitted resistance to integrase inhibitors was found in our population by Sanger sequencing or UDS.

Article number19752
JournalJournal of the International AIDS Society
Journal citation17 (Suppl 3)
ISSN1758-2652
Year2014
PublisherWiley
Publisher's version
License
CC BY 3.0
File Access Level
Open (open metadata and files)
Digital Object Identifier (DOI)https://doi.org/10.7448/ias.17.4.19752
PubMed ID25397497
Web address (URL)http://europepmc.org/abstract/med/25397497
Publication dates
Published02 Nov 2014

Related outputs

Dual production of polyhydroxyalkanoates and antibacterial/antiviral gold nanoparticles
McCormick, A.L., Basnett, P., Vecchiato, V., Wright, E., Noble, B., Marcello, E. and Paxinou, A. 2023. Dual production of polyhydroxyalkanoates and antibacterial/antiviral gold nanoparticles. Frontiers in Nanotechnology. 5 1243056. https://doi.org/10.3389/fnano.2023.1243056

Author Correction: A HML6 endogenous retrovirus on chromosome 3 is upregulated in amyotrophic lateral sclerosis motor cortex
Jones, A.R., Iacoangeli, A., Adey, B.N., Bowles, H., Shatunov, A., Troakes, C., Garson, J.A., McCormick, A.L. and Al‑Chalabi, A. 2022. Author Correction: A HML6 endogenous retrovirus on chromosome 3 is upregulated in amyotrophic lateral sclerosis motor cortex. Scientific Reports. 12 14171. https://doi.org/10.1038/s41598-022-18488-y

A HML6 endogenous retrovirus on chromosome 3 is upregulated in amyotrophic lateral sclerosis motor cortex.
Jones, A.R., Iacoangeli, A., Adey, B.N., Bowles, H., Shatunov, A, Troakes, C., Garson, J.A., McCormick, A.L. and Al‑Chalabi, A. 2021. A HML6 endogenous retrovirus on chromosome 3 is upregulated in amyotrophic lateral sclerosis motor cortex. Scientific Reports. 11 14283. https://doi.org/10.1038/s41598-021-93742-3

Elevated microRNA expression could be diagnostic biomarker for PDAC
Mortoglou, M., Arisan, D., Ferreira, T., McCormick, A.L. and Uysal Onganer, P. 2020. Elevated microRNA expression could be diagnostic biomarker for PDAC. Pancreatology. 20 (8) e19. https://doi.org/10.1016/j.pan.2020.10.015

Response to the Letter from Garcia-Montojo and colleagues concerning our paper entitled, Quantitative analysis of human endogenous retrovirus-K transcripts in postmortem premotor cortex fails to confirm elevated expression of HERV-K RNA in amyotrophic lateral sclerosis.
Garson, J.A., Usher, L., Al-Chalabi, A., Huggett, J., Day, E. and McCormick, A.L. 2019. Response to the Letter from Garcia-Montojo and colleagues concerning our paper entitled, Quantitative analysis of human endogenous retrovirus-K transcripts in postmortem premotor cortex fails to confirm elevated expression of HERV-K RNA in amyotrophic lateral sclerosis. Acta Neuropathologica Communications. 7 102. https://doi.org/10.1186/s40478-019-0756-9

Quantitative analysis of Human Endogenous Retrovirus-K transcripts in postmortem premotor cortex fails to confirm elevated expression of HERV-K RNA in amyotrophic lateral sclerosis
Garson, J.A., Usher, L., Al-Chalabi, A., Huggett, J., Day, E.F. and McCormick, A.L. 2019. Quantitative analysis of Human Endogenous Retrovirus-K transcripts in postmortem premotor cortex fails to confirm elevated expression of HERV-K RNA in amyotrophic lateral sclerosis. Acta Neuropathologica Communications. 7 45. https://doi.org/10.1186/s40478-019-0698-2

Correction: Evidence for Reduced Drug Susceptibility without Emergence of Major Protease Mutations following Protease Inhibitor Monotherapy Failure in the SARA Trial.
Sutherland, K.A., Parry, C.M., McCormick, A.L., Kapaata, A., Lyagoba, F., Kaleebu, P., Gilks, C.F., Goodall, R., Spyer, M., Kityo, C., Pillay, D., Gupta, R.K. and DART Virology Group 2016. Correction: Evidence for Reduced Drug Susceptibility without Emergence of Major Protease Mutations following Protease Inhibitor Monotherapy Failure in the SARA Trial. PLoS ONE. 11 (6) e0157094. https://doi.org/10.1371/journal.pone.0157094

Gag-Protease Sequence Evolution Following Protease Inhibitor Monotherapy Treatment Failure in HIV-1 Viruses Circulating in East Africa.
Sutherland, K.A., Goodall, R.L., McCormick, A.L., Kapaata, A., Lyagoba, F., Kaleebu, P., Thiltgen, G., Gilks, C.F., Spyer, M., Kityo, C., Pillay, D., Dunn, D. and Gupta, R.K. 2015. Gag-Protease Sequence Evolution Following Protease Inhibitor Monotherapy Treatment Failure in HIV-1 Viruses Circulating in East Africa. AIDS Research and Human Retroviruses. 31 (10), pp. 1032-1037. https://doi.org/10.1089/aid.2015.0138

Evidence for Reduced Drug Susceptibility without Emergence of Major Protease Mutations following Protease Inhibitor Monotherapy Failure in the SARA Trial
Sutherland, K.A., Parry, C.M., McCormick, A.L., Kapaata, A., Lyagoba, F., Kaleebu, P., Gilks, C.F., Goodall, R., Spyer, M., Kityo, C., Pillay, D., Gupta, R.K. and DART Virology Group 2015. Evidence for Reduced Drug Susceptibility without Emergence of Major Protease Mutations following Protease Inhibitor Monotherapy Failure in the SARA Trial. PLoS ONE. 10 (9) e0137834. https://doi.org/10.1371/journal.pone.0137834

IL28B genotype predicts response to chronic hepatitis C triple therapy with telaprevir or boceprevir in treatment naïve and treatment-experienced patients other than prior partial- and null-responders
Calisti, G., Tavares, A., Macartney, M.J., McCormick, A.L., Labbett, W., Jacobs, M., Dusheiko, G., Rosenberg, W.M. and Haque, T. 2015. IL28B genotype predicts response to chronic hepatitis C triple therapy with telaprevir or boceprevir in treatment naïve and treatment-experienced patients other than prior partial- and null-responders. SpringerPlus. 4 (1). https://doi.org/10.1186/s40064-015-1137-x

Evaluation of sequencing of HCV core/E1, NS5A and NS5B as a genotype predictive tool in comparison with commercial assays targeting 5′UTR
McCormick, A.L., Macartney, M.J., Abdi-Abshir, I., Labbett, W., Smith, C., Irish, D., Webster, D.P. and Haque, T. 2015. Evaluation of sequencing of HCV core/E1, NS5A and NS5B as a genotype predictive tool in comparison with commercial assays targeting 5′UTR. Journal of Clinical Virology. 66 (May), pp. 56-59. https://doi.org/10.1016/j.jcv.2015.03.006

Ultra-deep sequencing provides insights into the virology of hepatitis C super-infections in a case of three sequential infections with different genotypes
Chung, E., Ferns, R.B., He, M., Rigatti, R., Grant, P., McCormick, A.L., Bhagani, S., Webster, D.P., Nastouli, E. and Waters, L.J. 2015. Ultra-deep sequencing provides insights into the virology of hepatitis C super-infections in a case of three sequential infections with different genotypes. Journal of Clinical Virology. 70 (Sept), pp. 63-66. https://doi.org/10.1016/j.jcv.2015.06.105

The utility of genotypic tropism testing in clinical practice
Wyatt, H., Herman, O., Macartney, M.J., Conibear, T., Garcia-Diaz, A., Booth, C., McCormick, A.L., Smith, C., Johnson, M., Irish, D. and Webster, D. 2015. The utility of genotypic tropism testing in clinical practice. International Journal of STD and AIDS. 26 (8), pp. 593-594. https://doi.org/10.1177/0956462414546917

Phenotypic and genotypic analyses to guide selection of reverse transcriptase inhibitors in second-line HIV therapy following extended virological failure in Uganda
R. L. Goodall, D. T. Dunn, T. Pattery, A. van Cauwenberge, P. Nkurunziza, P. Awio, N. Ndembi, P. Munderi, C. Kityo, C. F. Gilks, P. Kaleebu, D. Pillay, P. Kaleebu, D. Pillay, P. Awio, M. Chirara, D. Dunn, D. M. Gibb, C. Gilks, R. Goodall, A. Kapaata, M. Katuramur, F. Lyagoba, R. Magala, B. Magambo, K. Mataruka, A.L. McCormick, L. Mugarura, T. Musunga, M. Nabankkema, J. Nkalubo, P. Nkurunziza, C. Parry, V. Robertson, M. Spyer, D. Yirrell, H. Grosskurth, P. Munderi, G. Kabuye, D. Nsibambi, R. Kasirye, E. Zalwango, M. Nakazibwe, B. Kikaire, G. Nassuna, R. Massa, K. Fadhiru, M. Namyalo, A. Zalwango, L. Generous and P. Khauka 2014. Phenotypic and genotypic analyses to guide selection of reverse transcriptase inhibitors in second-line HIV therapy following extended virological failure in Uganda. Journal of Antimicrobial Chemotherapy. 69 (7), pp. 1938-1944. https://doi.org/10.1093/jac/dku052

Feasibility of testing and detection of HIV-1 drug resistance in proviral DNA
Booth, C., McCormick, A.L., Garcia-Diaz, A., Macartney, M., Youle, M., Johnson, M. and Webster, D. 2014. Feasibility of testing and detection of HIV-1 drug resistance in proviral DNA. BMC Infectious Diseases. 14 (Supplement 4) O25. https://doi.org/10.1186/1471-2334-14-s4-o25

Telaprevir or boceprevir based therapy for chronic hepatitis C infection: Development of resistance-associated variants in treatment failure
Macartney, M.J., Irish, D., Bridge, S.H., Garcia-Diaz, A., Booth, C., McCormick, A.L., Labbett, W., Smith, C., Velazquez, C., Tanwar, S., Trembling, P., Jacobs, M., Dusheiko, G., Rosenberg, W.M. and Haque, T. 2014. Telaprevir or boceprevir based therapy for chronic hepatitis C infection: Development of resistance-associated variants in treatment failure. Antiviral Research. 105, pp. 112-7. https://doi.org/10.1016/j.antiviral.2014.02.019

Prevalence of baseline polymorphisms for potential resistance to NS5A inhibitors in drug-naive individuals infected with hepatitis C genotypes 1–4
McCormick, A.L., Wang, L., Garcia-Diaz, A., Macartney, M.J., Webster, D.P. and Haque, T. 2014. Prevalence of baseline polymorphisms for potential resistance to NS5A inhibitors in drug-naive individuals infected with hepatitis C genotypes 1–4 . Antiviral Therapy. 20 (1), pp. 81-85. https://doi.org/10.3851/IMP2763

The utility of different bioinformatics algorithms for genotypic HIV-1 tropism testing in a large clinical cohort with multiple subtypes
Bartlett, A., Macartney, M.J., Conibear, T., Feyertag, F., Smith, C., Johnson, M., Hyams, C., Garcia-Diaz, A., McCormick, A.L., Booth, C., Robertson, D. and Webster, D.P. 2014. The utility of different bioinformatics algorithms for genotypic HIV-1 tropism testing in a large clinical cohort with multiple subtypes. AIDS. 28 (11), pp. 1611-7. https://doi.org/10.1097/QAD.0000000000000288

Baseline drug-resistance mutations are detectable in HCV genes NS3 and NS5A but not NS5B in acute and chronic HIV-coinfected patients
McCormick, A.L., Moynihan, L., Macartney, M.J., Garcia-Diaz, A., Smith, C., Johnson, M., Rodger, A.J., Bhagani, S., Haque, T. and Webster, D. 2014. Baseline drug-resistance mutations are detectable in HCV genes NS3 and NS5A but not NS5B in acute and chronic HIV-coinfected patients. Antiviral Therapy. 20 (3), p. 2015. https://doi.org/10.3851/IMP2871

Evaluation of the Roche prototype 454 HIV-1 ultradeep sequencingdrug resistance assay in a routine diagnostic laboratory
Garcia-Diaz, A., Guerrero-Ramos, A., McCormick, A.L., Macartney, M.J., Conibear, T., Johnson, M., Haque, T. and Webster, D.P. 2013. Evaluation of the Roche prototype 454 HIV-1 ultradeep sequencingdrug resistance assay in a routine diagnostic laboratory. Journal of Clinical Virology. 58 (2), pp. 468-473. https://doi.org/10.1016/j.jcv.2013.07.009

Impact of the N348I Mutation in HIV-1 Reverse Transcriptase on Nonnucleoside Reverse Transcriptase Inhibitor Resistance in Non-Subtype B HIV-1
McCormick, A.L., Parry, C.M., Crombe, A., Goodall, R.L., Gupta, R.K., Kaleebu, P., Kityo, C., Chirara, M., Towers, G.J. and Pillay, D. 2011. Impact of the N348I Mutation in HIV-1 Reverse Transcriptase on Nonnucleoside Reverse Transcriptase Inhibitor Resistance in Non-Subtype B HIV-1. Antimicrobial Agents and Chemotherapy. 55 (4), pp. 1806-1809. https://doi.org/10.1128/AAC.01197-10

Full length HIV-1 Gag determines protease inhibitor susceptibility within in vitro assays
Gupta, R.K., Kohli, A., McCormick, A.L., Towers, G.J., Pillay, D. and Parry, C.M. 2010. Full length HIV-1 Gag determines protease inhibitor susceptibility within in vitro assays. AIDS. 24 (11), p. 1651–1655. https://doi.org/10.1097/QAD.0b013e3283398216

Lack of Minority K65RResistant Viral Populations Detected After Repeated Treatment Interruptions of Tenofovir/Zidovudine and Lamivudine in a Resource-Limited Setting
McCormick, A.L., Goodall, R.L., Joyce, A., Ndembi, N., Chirara, M., Katunduk, P., Walker, A.S., Yirrell, D.L., Gilks, C.F. and Pillay, D. 2010. Lack of Minority K65RResistant Viral Populations Detected After Repeated Treatment Interruptions of Tenofovir/Zidovudine and Lamivudine in a Resource-Limited Setting. Journal of Acquired Immune Deficiency Syndromes. 54 (2), pp. 215-216. https://doi.org/10.1097/QAI.0b013e3181cc1058

Viral Rebound and Emergence of Drug Resistance in the Absence of Viral Load Testing: A Randomized Comparison between Zidovudine-Lamivudine plus Nevirapine and Zidovudine-Lamivudine plus Abacavir
Ndembi, N., Goodall, R.L., Dunn, D.T., McCormick, A.L., Burke, A., Lyagoba, F., Munderi, P., Katundu, P., Kityo, C., Robertson, V., Yirrell, D.L., Walker, A.S., Gibb, D.M., Gilks, C.F., Kaleebu, P. and Pillay, D. 2010. Viral Rebound and Emergence of Drug Resistance in the Absence of Viral Load Testing: A Randomized Comparison between Zidovudine-Lamivudine plus Nevirapine and Zidovudine-Lamivudine plus Abacavir. The Journal of Infectious Diseases. 201 (1), pp. 106-113. https://doi.org/10.1086/648590

Quantification of reverse transcriptase in ALS and elimination of a novel retroviral candidate
McCormick, A.L., Brown, R.H., Cudkowicz, M.E., Al-Chalabi, A. and Garson, J.A. 2008. Quantification of reverse transcriptase in ALS and elimination of a novel retroviral candidate. Neurology. 70 (4), pp. 278-283. https://doi.org/10.1212/01.wnl.0000297552.13219.b4

An interferon gamma-gp120 fusion delivered as a DNA vaccine induces enhanced priming
Nimal, S., McCormick, A.L., Thomas, M.S. and Heath, A.W. 2005. An interferon gamma-gp120 fusion delivered as a DNA vaccine induces enhanced priming. Vaccine. 23 (10), pp. 3984-3990. https://doi.org/10.1016/j.vaccine.2005.01.160

CD40 antibody as an adjuvant induces enhanced T cell responses
Carlring, J., Barr, T.A., McCormick, A.L. and Heath, A.W. 2004. CD40 antibody as an adjuvant induces enhanced T cell responses. Vaccine. 22 (25-26), p. 3323–3328. https://doi.org/10.1016/j.vaccine.2004.02.043

A potent adjuvant effect of CD40 antibody attached to antigen
Barr, T.A., McCormick, A.L., Carlring, J. and Heath, A.W. 2003. A potent adjuvant effect of CD40 antibody attached to antigen. Immunology. 109 (1), pp. 87-92. https://doi.org/10.1046/j.1365-2567.2003.01634.x

Immunization with an Interferon-g–gp120 Fusion Protein Induces Enhanced Immune Responses to Human Immunodeficiency Virus gp120
McCormick, A.L., Thomas, M.S. and Heath, A.W. 2001. Immunization with an Interferon-g–gp120 Fusion Protein Induces Enhanced Immune Responses to Human Immunodeficiency Virus gp120. The Journal of Infectious Diseases. 184 (11), pp. 1423-1430. https://doi.org/10.1086/324371

Gag Determinants of Fitness and Drug Susceptibility in Protease Inhibitor-Resistant Human Immunodeficiency Virus Type 1
Parry, C.M., Kohli, A., Boinett, C.J., Towers, G.J., McCormick, A.L. and Pillay, D. 1999. Gag Determinants of Fitness and Drug Susceptibility in Protease Inhibitor-Resistant Human Immunodeficiency Virus Type 1. Journal of Virology. 83 (18), pp. 9094-9101. https://doi.org/10.1128/JVI.02356-08

Cell Surface Expression of CD154 Inhibits Alloantibody Responses: A Mechanism for the Prevention of Autoimmune Responses against Activated T Cells?
McCormick, A.L., Santos-Argumedo, L., Thomas, M.S. and Heath, A.W. 1999. Cell Surface Expression of CD154 Inhibits Alloantibody Responses: A Mechanism for the Prevention of Autoimmune Responses against Activated T Cells? Cellular Immunology. 195 (2), pp. 157-161. https://doi.org/10.1006/cimm.1999.1528

Enhancement of immunogenicity of recombinant antigens by production of a cytokine-antigen fusion protein for vaccination
McCormick, A.L., Thomas, M.S. and Heath, A.W. 1997. Enhancement of immunogenicity of recombinant antigens by production of a cytokine-antigen fusion protein for vaccination. Biochemical Society Transactions. 25, p. 297S. https://doi.org/10.1042/bst025297s

Permalink - https://westminsterresearch.westminster.ac.uk/item/w56y8/analysis-of-transmitted-hiv-1-drug-resistance-using-454-ultra-deep-sequencing-and-the-deepchek-hiv-system


Share this

Usage statistics

19 total views
15 total downloads
These values cover views and downloads from WestminsterResearch and are for the period from September 2nd 2018, when this repository was created.