Laboratory Guidelines for the Practical Use of HIV Drug Resistance Tests in Patient Follow-Up

HIV drug resistance is one of the major limitations in the successful treatment of HIV-infected patients using currently available antiretroviral combination therapies. When appropriate, drug susceptibility profiles should be taken into consideration in the choice of a specific combination therapy. Guidelines recommending resistance testing in certain circumstances have been issued. Many clinicians have access to resistance testing and will increasingly use these results in their treatment decisions. In this document, we comment on the different methods available, and the relevant issues relating to the clinical application of these tests. Specifically, the following recommendations can be made: (i) genotypic and phenotypic HIV-1 drug resistance analyses can yield complementary information for the clinician. However, insufficient information currently exists as to which approach is preferable in any particular clinical setting; (ii) when HIV-1 drug resistance testing is required, it is recommended that testing be performed on plasma samples obtained before starting, stopping or changing therapy, on samples that have a viral load above the detection limit of the resistance test; (iii) the panel recommends that genotypic and phenotypic HIV-1 drug resistance testing for clinical purposes be performed in a certified laboratory under strict quality control and quality assurance standards; and (iv) the panel recommends that resistance testing laboratories provide clinicians with resistance reports that include a list of drug-related resistance mutations (genotype) and/or a list of drug-related fold resistance values (phenotype), with interpretations of each by an experienced virologist. The interpretation of genotypic and phenotypic analysis is a complex and developing science, and in order to understand HIV-1 drug resistance reports, communication between the requesting clinician and the expert that interpreted the resistance report is recommended.

[1]  E. De Clercq,et al.  Evaluation of two commercial kits for the detection of genotypic drug resistance on a panel of HIV type 1 subtypes A through J. , 2001, Journal of acquired immune deficiency syndromes.

[2]  K. Hertogs,et al.  Clinical and laboratory guidelines for the use of HIV-1 drug resistance testing as part of treatment management: recommendations for the European setting , 2001 .

[3]  Barbara Schmidt,et al.  Simple algorithm derived from a geno-/phenotypic database to predict HIV-1 protease inhibitor resistance , 2000, AIDS.

[4]  D. Persing,et al.  Comparative Evaluation of Three Human Immunodeficiency Virus Genotyping Systems: the HIV-GenotypR Method, the HIV PRT GeneChip Assay, and the HIV-1 RT Line Probe Assay , 2000, Journal of Clinical Microbiology.

[5]  J. Schapiro,et al.  Importance of protease inhibitor plasma levels in HIV-infected patients treated with genotypic-guided therapy: pharmacological data from the Viradapt Study , 2000, AIDS.

[6]  V. Miller, PhD European guidelines on resistance testing: why are they needed? , 2000 .

[7]  T. Merigan,et al.  A randomized study of antiretroviral management based on plasma genotypic antiretroviral resistance testing in patients failing therapy , 2000 .

[8]  S. Hammer,et al.  Antiretroviral drug resistance testing in adult HIV-1 infection: recommendations of an International AIDS Society-USA Panel. , 2000, JAMA.

[9]  E. De Clercq,et al.  Baseline HIV type 1 genotypic resistance to a newly added nucleoside analog is predictive of virologic failure of the new therapy. , 2000, AIDS research and human retroviruses.

[10]  Christos J. Petropoulos,et al.  A Novel Phenotypic Drug Susceptibility Assay for Human Immunodeficiency Virus Type 1 , 2000, Antimicrobial Agents and Chemotherapy.

[11]  E. De Clercq,et al.  Patient HIV-1 strains carrying the multiple nucleoside resistance mutations are cross-resistant to abacavir. , 2000, AIDS.

[12]  V. Soriano,et al.  Emergence of zidovudine resistance in HIV-infected patients receiving stavudine. , 2000, Journal of acquired immune deficiency syndromes.

[13]  M. Hirsch,et al.  Drug susceptibility in HIV infection after viral rebound in patients receiving indinavir-containing regimens. , 2000, JAMA.

[14]  V. Calvez,et al.  Mechanisms of virologic failure in previously untreated HIV-infected patients from a trial of induction-maintenance therapy. Trilège (Agence Nationale de Recherches sur le SIDA 072) Study Team). , 2000, JAMA.

[15]  K. Hertogs,et al.  HIV Drug Susceptibility and Treatment Response to Mega-Haart Regimen in Patients from the Frankfurt HIV Cohort , 2000, Antiviral therapy.

[16]  S. Hammer,et al.  The Relation between Baseline HIV Drug Resistance and Response to Antiretroviral Therapy: Re-Analysis of Retrospective and Prospective Studies Using a Standardized Data Analysis Plan , 2000, Antiviral therapy.

[17]  J. Schapiro,et al.  Persisting Long-Term Benefit of Genotype-Guided Treatment for HIV-Infected Patients Failing Haart. The Viradapt Study: Week 48 Follow-Up , 2000, Antiviral therapy.

[18]  C. Verhofstede,et al.  Interruption of reverse transcriptase inhibitors or a switch from reverse transcriptase to protease inhibitors resulted in a fast reappearance of virus strains with a reverse transcriptase inhibitor-sensitive genotype. , 1999, AIDS.

[19]  M. Youle,et al.  Rapid decline in detectability of HIV-1 drug resistance mutations after stopping therapy. , 1999, AIDS.

[20]  C. Boucher,et al.  Increased fitness of drug resistant HIV-1 protease as a result of acquisition of compensatory mutations during suboptimal therapy. , 1999, AIDS.

[21]  P. Harrigan,et al.  Baseline HIV drug resistance profile predicts response to ritonavir-saquinavir protease inhibitor therapy in a community setting. , 1999, AIDS.

[22]  D. Richman,et al.  Sequence clusters in human immunodeficiency virus type 1 reverse transcriptase are associated with subsequent virological response to antiretroviral therapy. , 1999, The Journal of infectious diseases.

[23]  D. Ho,et al.  Phenotypic changes in drug susceptibility associated with failure of human immunodeficiency virus type 1 (HIV-1) triple combination therapy. , 1999, The Journal of infectious diseases.

[24]  B. Larder,et al.  Performance of the Affymetrix GeneChip HIV PRT 440 Platform for Antiretroviral Drug Resistance Genotyping of Human Immunodeficiency Virus Type 1 Clades and Viral Isolates with Length Polymorphisms , 1999, Journal of Clinical Microbiology.

[25]  D. Stuart,et al.  A Family of Insertion Mutations between Codons 67 and 70 of Human Immunodeficiency Virus Type 1 Reverse Transcriptase Confer Multinucleoside Analog Resistance , 1999, Antimicrobial Agents and Chemotherapy.

[26]  J. Eron,et al.  Antiretroviral-Drug Concentrations in Semen: Implications for Sexual Transmission of Human Immunodeficiency Virus Type 1 , 1999, Antimicrobial Agents and Chemotherapy.

[27]  M. Kazatchkine,et al.  Efficacy of a five-drug combination including ritonavir, saquinavir and efavirenz in patients who failed on a conventional triple-drug regimen: phenotypic resistance to protease inhibitors predicts outcome of therapy. , 1999, AIDS.

[28]  C. Boucher,et al.  Worldwide Evaluation of DNA Sequencing Approaches for Identification of Drug Resistance Mutations in the Human Immunodeficiency Virus Type 1 Reverse Transcriptase , 1999, Journal of Clinical Microbiology.

[29]  J. Schapiro,et al.  Drug-resistance genotyping in HIV-1 therapy: the VIRAD APT randomi sed controlled trial , 1999, The Lancet.

[30]  B. Schmidt,et al.  Rapid, phenotypic HIV-1 drug sensitivity assay for protease and reverse transcriptase inhibitors. , 1999, Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology.

[31]  P. Volberding,et al.  Novel four-drug salvage treatment regimens after failure of a human immunodeficiency virus type 1 protease inhibitor-containing regimen: antiviral activity and correlation of baseline phenotypic drug susceptibility with virologic outcome. , 1999, The Journal of infectious diseases.

[32]  E. Acosta,et al.  Indinavir Concentrations and Antiviral Effect , 1999, Pharmacotherapy.

[33]  M. Zazzi,et al.  Genotypic Resistance to Zidovudine as a Predictor of Failure of Subsequent Therapy with Human Immunodeficiency Virus Type-1 Nucleoside Reverse-Transcriptase Inhibitors , 1999, European Journal of Clinical Microbiology and Infectious Diseases.

[34]  J. Lisziewicz,et al.  Analysis of amino insertion mutations in the fingers subdomain of HIV-1 reverse transcriptase. , 1999, Journal of molecular biology.

[35]  C. Mandl,et al.  Comparison of line probe assay (LIPA) and sequence analysis for detection of HIV‐1 drug resistance , 1999, Journal of medical virology.

[36]  M. Youle,et al.  HIV-1 Drug Resistance Testing: Health Outcomes Issues , 1999, Antiviral therapy.

[37]  D. Pieniążek,et al.  Genetic Variation and Susceptibilities to Protease Inhibitors among Subtype B and F Isolates in Brazil , 1999, Antimicrobial Agents and Chemotherapy.

[38]  M. Hillebrand,et al.  Insertion of two amino acids combined with changes in reverse transcriptase containing tyrosine-215 of HIV-1 resistant to multiple nucleoside analogs. , 1999, AIDS.

[39]  V. Calvez,et al.  Stavudine Resistance: An Update on Susceptibility following Prolonged Therapy , 1999, Antiviral therapy.

[40]  D. Katzenstein,et al.  A 6-basepair insert in the reverse transcriptase gene of human immunodeficiency virus type 1 confers resistance to multiple nucleoside inhibitors. , 1998, The Journal of clinical investigation.

[41]  P. Vernazza,et al.  Resistance of HIV‐1 to antiretroviral agents in blood and seminal plasma: implications for transmission , 1998, AIDS.

[42]  D. R. Kuritzkes,et al.  Genotypic and Phenotypic Characterization of Human Immunodeficiency Virus Type 1 Variants Isolated from Patients Treated with the Protease Inhibitor Nelfinavir , 1998, Antimicrobial Agents and Chemotherapy.

[43]  J. Fantini,et al.  Stable rearrangements of the β3–β4 hairpin loop of HIV‐1 reverse transcriptase in plasma viruses from patients receiving combination therapy , 1998, AIDS.

[44]  T. Harrer,et al.  Multiple dideoxynucleoside analogue‐resistant (MddNR) HIV‐1 strains isolated from patients from different European countries , 1998, AIDS.

[45]  Dale J. Kempf,et al.  In Vitro Selection and Characterization of Human Immunodeficiency Virus Type 1 Variants with Increased Resistance to ABT-378, a Novel Protease Inhibitor , 1998, Journal of Virology.

[46]  J W Mulder,et al.  The effect of plasma drug concentrations on HIV‐1 clearance rate during quadruple drug therapy , 1998, AIDS.

[47]  E. De Clercq,et al.  Comparison of the LiPA HIV-1 RT test, selective PCR and direct solid phase sequencing for the detection of HIV-1 drug resistance mutations. , 1998, Journal of virological methods.

[48]  D. Richman,et al.  Comparative performance of high-density oligonucleotide sequencing and dideoxynucleotide sequencing of HIV type 1 pol from clinical samples. , 1998, AIDS Research and Human Retroviruses.

[49]  T. Merigan,et al.  Multiple Concurrent Reverse Transcriptase and Protease Mutations and Multidrug Resistance of HIV-1 Isolates from Heavily Treated Patients , 1998, Annals of Internal Medicine.

[50]  L. Pérez-Alvárez,et al.  Evaluation of Mixtures of Wild-Type HIV-1 and HIV-1 with Resistance Point Mutations against Reverse Transcriptase Inhibitors , 1998, Antiviral therapy.

[51]  F. Brun-Vézinet,et al.  Human Immunodeficiency Virus Type 1 Subtype F Reverse Transcriptase Sequence and Drug Susceptibility , 1998, Journal of Virology.

[52]  F. Mammano,et al.  Loss of Viral Fitness Associated with Multiple Gag and Gag-Pol Processing Defects in Human Immunodeficiency Virus Type 1 Variants Selected for Resistance to Protease Inhibitors In Vivo , 1998, Journal of Virology.

[53]  L. Fitzgerald,et al.  Human immunodeficiency virus in semen arises from a genetically distinct virus reservoir. , 1998, AIDS research and human retroviruses.

[54]  D. Richman,et al.  Human Immunodeficiency Virus Replication and Genotypic Resistance in Blood and Lymph Nodes after a Year of Potent Antiretroviral Therapy , 1998, Journal of Virology.

[55]  D. Katzenstein,et al.  HIV-1 reverse transcriptase codon 215 mutation in plasma RNA: immunologic and virologic responses to zidovudine. The AIDS Clinical Trials Group Study 175 Virology Team. , 1998, Journal of acquired immune deficiency syndromes and human retrovirology : official publication of the International Retrovirology Association.

[56]  E. De Clercq,et al.  Evolution of HIV drug Resistance in Zidovudine/Zalcitabine- and Zidovudine/ Didanosine-Experienced Patients Receiving Lamivudine-Containing Combination Therapy , 1998 .

[57]  Brendan Larder,et al.  A Rapid Method for Simultaneous Detection of Phenotypic Resistance to Inhibitors of Protease and Reverse Transcriptase in Recombinant Human Immunodeficiency Virus Type 1 Isolates from Patients Treated with Antiretroviral Drugs , 1998, Antimicrobial Agents and Chemotherapy.

[58]  J. Albert,et al.  Drug susceptibility of subtypes A,B,C,D, and E human immunodeficiency virus type 1 primary isolates. , 1998, AIDS research and human retroviruses.

[59]  C. Crumpacker,et al.  Attenuated replication of human immunodeficiency virus type 1 with a didanosine-selected reverse transcriptase mutation , 1997, Journal of virology.

[60]  F. Brun-Vézinet,et al.  Susceptibility of human immunodeficiency virus type 1 group O isolates to antiretroviral agents: in vitro phenotypic and genotypic analyses , 1997, Journal of virology.

[61]  D. Leduc,et al.  Toxicity, efficacy, plasma drug concentrations and protease mutations in patients with advanced HIV infection treated with ritonavir plus saquinavir , 1997, AIDS.

[62]  E A Emini,et al.  Treatment with indinavir, zidovudine, and lamivudine in adults with human immunodeficiency virus infection and prior antiretroviral therapy. , 1997, The New England journal of medicine.

[63]  Tom de Groot,et al.  Lamivudine-resistant human immunodeficiency virus type 1 variants (184V) require multiple amino acid changes to become co-resistant to zidovudine in vivo. , 1997, The Journal of infectious diseases.

[64]  R. Bhikhabhai,et al.  A Non-Radioactive Microtitre Plate Reverse Transcriptase (RT) Assay, Based on Immobilized Template, for Screening of RT Activity Inhibitors and Evaluation of their Mode of Action , 1997 .

[65]  L. Stuyver,et al.  Line probe assay for rapid detection of drug-selected mutations in the human immunodeficiency virus type 1 reverse transcriptase gene , 1997, Antimicrobial agents and chemotherapy.

[66]  E. De Clercq,et al.  Multiple drug resistance to nucleoside analogues and nonnucleoside reverse transcriptase inhibitors in an efficiently replicating human immunodeficiency virus type 1 patient strain. , 1996, The Journal of infectious diseases.

[67]  D. Mayers Rational approaches to resistance: nucleoside analogues. , 1996, AIDS.

[68]  S. Hammer,et al.  The relation of virologic and immunologic markers to clinical outcomes after nucleoside therapy in HIV-infected adults with 200 to 500 CD4 cells per cubic millimeter. AIDS Clinical Trials Group Study 175 Virology Study Team. , 1996, The New England journal of medicine.

[69]  J. Mellors,et al.  Zidovudine resistance is suppressed by mutations conferring resistance of human immunodeficiency virus type 1 to foscarnet , 1996, Journal of virology.

[70]  W. Keulen,et al.  Reduced replication of 3TC‐resistant HIV‐1 variants in primary cells due to a processivity defect of the reverse transcriptase enzyme. , 1996, The EMBO journal.

[71]  J. Gambertoglio,et al.  Zidovudine phosphorylation after short‐term and long‐term therapy with zidovudine in patients infected with the human immunodeficiency virus , 1996, Clinical pharmacology and therapeutics.

[72]  R. Shafer,et al.  Interlaboratory comparison of sequence-specific PCR and ligase detection reaction to detect a human immunodeficiency virus type 1 drug resistance mutation. The AIDS Clinical Trials Group Virology Committee Drug Resistance Working Group , 1996, Journal of clinical microbiology.

[73]  D. Ho,et al.  Ordered accumulation of mutations in HIV protease confers resistance to ritonavir , 1996, Nature Medicine.

[74]  J. Metcalf,et al.  Randomized, controlled phase I/II, trial of combination therapy with delavirdine (U-90152S) and conventional nucleosides in human immunodeficiency virus type 1-infected patients , 1996, Antimicrobial agents and chemotherapy.

[75]  Douglas Richman,et al.  Viral Dynamics of HIV: Implications for Drug Development and Therapeutic Strategies , 1996, Annals of Internal Medicine.

[76]  D. Faulds,et al.  Stavudine: a review of its pharmacodynamic and pharmacokinetic properties and clinical potential in HIV infection. , 1996, Drugs.

[77]  A. Caliendo,et al.  Effects of zidovudine-selected human immunodeficiency virus type 1 reverse transcriptase amino acid substitutions on processive DNA synthesis and viral replication , 1996, Journal of virology.

[78]  T. Merigan,et al.  Multidrug-resistant human immunodeficiency virus type 1 strains resulting from combination antiretroviral therapy , 1996, Journal of virology.

[79]  T. Merigan,et al.  The prognostic significance of serum viral load, codon 215 reverse transcriptase mutation and CD4+ T cells on progression of HIV disease in a double-blind study of thymopentin , 1996, AIDS.

[80]  R. Nuesch,et al.  The Fas receptor in HIV infection: expression on peripheral blood lymphocytes and role in the depletion of T cells , 1996, AIDS.

[81]  D. Ho,et al.  A preliminary study of ritonavir, an inhibitor of HIV-1 protease, to treat HIV-1 infection. , 1995, The New England journal of medicine.

[82]  L. M. Lehman,et al.  A short-term study of the safety, pharmacokinetics, and efficacy of ritonavir, an inhibitor of HIV-1 protease. European-Australian Collaborative Ritonavir Study Group. , 1995, The New England journal of medicine.

[83]  H. Mitsuya,et al.  Enzymatic Characterization of Human Immunodeficiency Virus Type 1 Reverse Transcriptase Resistant to Multiple 2′,3′-Dideoxynucleoside 5′-Triphosphates (*) , 1995, The Journal of Biological Chemistry.

[84]  C. Loveday,et al.  The appearance of drug resistance-associated point mutations in HIV type 1 plasma RNA precedes their appearance in proviral DNA. , 1995, AIDS research and human retroviruses.

[85]  E. Furfine,et al.  Analysis of resistance to human immunodeficiency virus type 1 protease inhibitors by using matched bacterial expression and proviral infection vectors , 1995, Journal of virology.

[86]  S D Kemp,et al.  Potential mechanism for sustained antiretroviral efficacy of AZT-3TC combination therapy. , 1995, Science.

[87]  M. Wainberg,et al.  Host Cell Dependence of Human Immunodeficiency Virus Type-1 Drug Resistance Profiles and Tissue Culture Selection Patterns , 1995 .

[88]  M. Magnani,et al.  2',3'-Dideoxycytidine induced drug resistance in human cells. , 1995, Life sciences.

[89]  P. Boyer,et al.  Analysis of mutations at position 184 in reverse transcriptase of human immunodeficiency virus type 1 , 1995, Antimicrobial agents and chemotherapy.

[90]  J. Sninsky,et al.  Rapid changes in human immunodeficiency virus type 1 RNA load and appearance of drug-resistant virus populations in persons treated with lamivudine (3TC). , 1995, The Journal of infectious diseases.

[91]  E. Arnold,et al.  Emergence of human immunodeficiency virus type 1 variants with resistance to multiple dideoxynucleosides in patients receiving therapy with dideoxynucleosides. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[92]  T. Greenough,et al.  High-dose nevirapine: safety, pharmacokinetics, and antiviral effect in patients with human immunodeficiency virus infection. , 1995, The Journal of infectious diseases.

[93]  J. Albert,et al.  Comparison of the sensitivities of primary isolates of HIV type 2 and HIV type 1 to antiviral drugs and drug combinations. , 1994, AIDS Research and Human Retroviruses.

[94]  O. Turriziani,et al.  Zidovudine induces the expression of cellular resistance affecting its antiviral activity. , 1994, AIDS research and human retroviruses.

[95]  D. Katzenstein,et al.  Combination therapy with zidovudine and didanosine selects for drug-resistant human immunodeficiency virus type 1 strains with unique patterns of pol gene mutations. , 1994, The Journal of infectious diseases.

[96]  B. Larder,et al.  Recombinant virus assay: a rapid, phenotypic assay for assessment of drug susceptibility of human immunodeficiency virus type 1 isolates , 1994, Antimicrobial Agents and Chemotherapy.

[97]  E. De Clercq,et al.  Treatment of human immunodeficiency virus type 1 (HIV-1)-infected cells with combinations of HIV-1-specific inhibitors results in a different resistance pattern than does treatment with single-drug therapy , 1993, Journal of virology.

[98]  I. Chen,et al.  A mutation in reverse transcriptase of bis(heteroaryl)piperazine-resistant human immunodeficiency virus type 1 that confers increased sensitivity to other nonnucleoside inhibitors. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[99]  B. Larder 3'-Azido-3'-deoxythymidine resistance suppressed by a mutation conferring human immunodeficiency virus type 1 resistance to nonnucleoside reverse transcriptase inhibitors , 1992, Antimicrobial Agents and Chemotherapy.

[100]  C. Loveday,et al.  A microtitre format point mutation assay: Application to the detection of drug resistance in human immunodeficiency virus type‐1 infected patients treated with zidovudine , 1992, Journal of medical virology.

[101]  S D Kemp,et al.  Resistance to ddI and sensitivity to AZT induced by a mutation in HIV-1 reverse transcriptase. , 1991, Science.

[102]  S D Kemp,et al.  Zidovudine resistance predicted by direct detection of mutations in DNA from HIV-infected lymphocytes. , 1991, AIDS.

[103]  J Desmyter,et al.  Evaluating Clinical Isolates for Their Phenotypic and Genotypic Resistance Against Anti-HIV Drugs. , 2000, Methods in molecular medicine.

[104]  A. Vandamme,et al.  Multidrug resistant human immunodeficiency virus type I , 2000 .

[105]  Rami Kantor,et al.  The Genetic Basis of HIV-1 Resistance to Reverse Transcriptase and Protease Inhibitors. , 2000, AIDS reviews.

[106]  Bryan Chan,et al.  Human immunodeficiency virus reverse transcriptase and protease sequence database , 2003, Nucleic Acids Res..

[107]  D. Katzenstein,et al.  Sequence and drug susceptibility of subtype C protease from human immunodeficiency virus type 1 seroconverters in Zimbabwe. , 1999, AIDS research and human retroviruses.

[108]  E. De Clercq,et al.  Managing resistance to anti-HIV drugs: an important consideration for effective disease management. , 1999, Drugs.

[109]  A. Bertoli,et al.  Clinical implications of HIV dynamics and drug resistance in macrophages. , 1998, Journal of biological regulators and homeostatic agents.

[110]  B. Larder,et al.  Mutations in Retroviral Genes Associated with Drug Resistance , 1996 .

[111]  S. Emery,et al.  A Review of its Pharmacodynamic and Pharmacokinetic Properties and Clinical Potential in HIV Infection , 1996 .

[112]  H. Nitschko,et al.  Identification, purification, and cell culture assays of retroviral proteases. , 1994, Methods in enzymology.

[113]  B. Gazzard,et al.  Characterization of HIV-1 strains isolated from patients treated with TIBO R82913. , 1994, AIDS research and human retroviruses.

[114]  J. Balzarini,et al.  Biochemical pharmacology of nucleoside analogues active against HIV , 1994 .

[115]  B. Larder,et al.  Ordered appearance of zidovudine resistance mutations during treatment of 18 human immunodeficiency virus-positive subjects. , 1992, The Journal of infectious diseases.