“All-in-One Assay”, a direct phenotypic anti-human immunodeficiency virus type 1 drug resistance assay for three-drug combination therapies that takes into consideration in vivo drug concentrations

Hachiya, Atsuko; Matsuoka-Aizawa, Saori; Tsuchiya, Kiyoto; Gatanaga, Hiroyuki; Kimura, Satoshi; Tatsumi, Masashi; Oka, Shinichi

doi:10.1016/s0166-0934(03)00150-2

Cited by 6 publications

(4 citation statements)

References 37 publications

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“…EFV, NVP, and ETV were generously provided by Merck Co., Inc. (Rahway, NJ), Boehringer Ingelheim Pharmaceutics Inc. (Ridgefield, CT), and Tibotec Pharmaceuticals (Little Island, Co., Cork, Ireland), respectively. Recombinant and isolated HIV-1 susceptibility to EFV, NVP, and ETV was determined in triplicate using MAGIC-5 cells (10,12,16). Briefly, MAGIC-5 cells were infected with an adjusted virus stock (300 BFU) in various concentrations of NNRTIs, cultured for 48 h, fixed, and stained with 5-bromo-4-chloro-3-indolyl-␤-D-galactopyranoside (Takara Shuzo, Ohtsu, Japan).…”

Section: Hiv-1 Sequences and Clinical Isolates From Treatment-naïve Imentioning

confidence: 99%

Combination of V106I and V179D Polymorphic Mutations in Human Immunodeficiency Virus Type 1 Reverse Transcriptase Confers Resistance to Efavirenz and Nevirapine but Not Etravirine

Gatanaga

Ode

Hachiya

et al. 2010

Antimicrob Agents Chemother

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Etravirine (ETV) is a second-generation nonnucleoside reverse transcriptase (RT) inhibitor (NNRTI)introduced recently for salvage antiretroviral treatment after the emergence of NNRTI-resistant human immunodeficiency virus type 1 (HIV-1). Following its introduction, two naturally occurring mutations in HIV-1 RT, V106I and V179D, were listed as ETV resistance-associated mutations. However, the effect of these mutations on the development of NNRTI resistance has not been analyzed yet. To select highly NNRTIresistant HIV-1 in vitro, monoclonal HIV-1 strains harboring V106I and V179D (HIV-1 V106I and HIV-1 V179D ) were propagated in the presence of increasing concentrations of efavirenz (EFV). Interestingly, V179D emerged in one of three selection experiments from HIV-1 V106I and V106I emerged in two of three experiments from HIV-1 V179D . Analysis of recombinant HIV-1 clones showed that the combination of V106I and V179D conferred significant resistance to EFV and nevirapine (NVP) but not to ETV. Structural analysis indicated that ETV can overcome the repulsive interactions caused by the combination of V106I and V179D through fine-tuning of its binding module to RT facilitated by its plastic structure, whereas EFV and NVP cannot because of their rigid structures. Analysis of clinical isolates showed comparable drug susceptibilities, and the same combination of mutations was found in some database patients who experienced virologic NNRTI-based treatment failure. The combination of V106I and V179D is a newly identified NNRTI resistance pattern of mutations. The combination of polymorphic and minor resistance-associated mutations should be interpreted carefully.

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Section: Hiv-1 Sequences and Clinical Isolates From Treatment-naïve Imentioning

confidence: 99%

Combination of V106I and V179D Polymorphic Mutations in Human Immunodeficiency Virus Type 1 Reverse Transcriptase Confers Resistance to Efavirenz and Nevirapine but Not Etravirine

Gatanaga

Ode

Hachiya

et al. 2010

Antimicrob Agents Chemother

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“…The former uses a chimeric virus generated by homologous recombination of RT-PR PCR-derived sequences, which is then cultured with cells and different drug concentrations [10, 11]. The latter uses PBMCs co-culture to isolate HIV-1 and is then incubated in MAGIC-5 cells with different drug concentrations [12, 13]. It has been demonstrated that the DR mutations can impair HIV-1 fitness [36–40].…”

Section: Discussionmentioning

confidence: 99%

“…However, several studies have demonstrated that T-cell-tropic (X4) and macrophage-tropic (R5) viruses can effectively produce using MAGIC-5 cells. Therefore, both R5-tropic and X4-tropic viruses can using MAGIC-5 cells to determine the HIV-1 drug susceptibility to PIs and RTIs [12, 13]. …”

Section: Discussionmentioning

confidence: 99%

“…In contrast, phenotypic assays measure HIV-1 viral replication in cells cultured in different drug concentrations. There are two types of phenotypic assays: commercially available phenotypic assays generate chimeric viruses by homologous recombination of PCR-derived sequences and then culture with cells in different drug concentrations [10, 11] and in-house phenotypic assay use peripheral blood mononuclear cells (PBMCs) to isolate HIV-1 and then incubate them in target cells (MAGIC-5 cells) with different drug concentrations [12, 13]. It has been reported that phenotypic drug resistance using recombinant virus assay was limited to detect low-frequency viral quasispecies below than 50% [14].…”

Section: Introductionmentioning

confidence: 99%

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Characterization of the Drug Resistance Profiles of Patients Infected with CRF07_BC Using Phenotypic Assay and Ultra-Deep Pyrosequencing

Huang

Wang

et al. 2017

PLoS ONE

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The usefulness of ultra-deep pyrosequencing (UDPS) for the diagnosis of HIV-1 drug resistance (DR) remains to be determined. Previously, we reported an explosive outbreak of HIV-1 circulating recombinant form (CRF) 07_BC among injection drug users (IDUs) in Taiwan in 2004. The goal of this study was to characterize the DR of CRF07_BC strains using different assays including UDPS. Seven CRF07_BC isolates including 4 from early epidemic (collected in 2004–2005) and 3 from late epidemic (collected in 2008) were obtained from treatment-naïve patient’s peripheral blood mononuclear cells. Viral RNA was extracted directly from patient’s plasma or from cultural supernatant and the pol sequences were determined using RT-PCR sequencing or UDPS. For comparison, phenotypic drug susceptibility assay using MAGIC-5 cells (in-house phenotypic assay) and Antivirogram were performed. In-house phenotypic assay showed that all the early epidemic and none of the late epidemic CRF07_BC isolates were resistant to most protease inhibitors (PIs) (4.4–47.3 fold). Neither genotypic assay nor Antivirogram detected any DR mutations. UDPS showed that early epidemic isolates contained 0.01–0.08% of PI DR major mutations. Furthermore, the combinations of major and accessory PI DR mutations significantly correlated with the phenotypic DR. The in-house phenotypic assay is superior to other conventional phenotypic assays in the detection of DR variants with a frequency as low as 0.01%.

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Laboratory Testing for HIV Infection

Shepherd

Laeyendecker²,

Quinn³

2008

Global HIV/AIDS Medicine

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“All-in-One Assay”, a direct phenotypic anti-human immunodeficiency virus type 1 drug resistance assay for three-drug combination therapies that takes into consideration in vivo drug concentrations

Cited by 6 publications

References 37 publications

Combination of V106I and V179D Polymorphic Mutations in Human Immunodeficiency Virus Type 1 Reverse Transcriptase Confers Resistance to Efavirenz and Nevirapine but Not Etravirine

Combination of V106I and V179D Polymorphic Mutations in Human Immunodeficiency Virus Type 1 Reverse Transcriptase Confers Resistance to Efavirenz and Nevirapine but Not Etravirine

Characterization of the Drug Resistance Profiles of Patients Infected with CRF07_BC Using Phenotypic Assay and Ultra-Deep Pyrosequencing

Laboratory Testing for HIV Infection

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