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  • 1
    In: Future Virology, Future Medicine Ltd, Vol. 8, No. 8 ( 2013-08), p. 815-820
    Abstract: Aim: The objective of the present study was to investigate whether patients with transmitted resistance more frequently harbor X4/DM tropic viral strains. Patients & methods: Patients were included from an Italian nationwide database if they were tested for tropism and resistance at the same time. HIV tropism was assessed by the Geno2pheno coreceptor system (false-positive rate: ≤10%) and enhanced-sensitivity Trofile assay. Overall, 299 naive patients, tested between 2009 and 2011, were included: 252 patients tested by Geno2pheno, 116 by enhanced-sensitivity Trofile assay and 80 by both methods. Results & conclusion: Using Geno2pheno, X4/DM tropic virus was detected in 55 patients (21.8%), with an overall mean false-positive rate of 42.3% (standard deviation: ±33.3). Using the enhanced-sensitivity Trofile assay, 29 patients (25.0%) carried X4/DM tropic virus. Resistance mutations were more frequently detected in patients harboring X4/DM tropic virus (mean: 1.18 ± 3.0 vs 0.41 ± 1.2 per patient; p = 0.001) and with both Geno2pheno (0.82 ± 2.6 vs 0.35 ± 0.9; p = 0.034) and enhanced-sensitivity Trofile assay (1.11 ± 1.9 vs 0.46 ± 1.1; p = 0.039). However, significant differences were found for reverse transcriptase-related mutations, but not for transmitted protease resistance, and this might be explained by the low frequency of transmitted protease resistance. Among single mutations, L33F and L90M with regards to protease and K65R, K70E, K219E and V106A/M with regards to reverse transcriptase were found to be significantly associated with X4/DM tropic virus. X4/DM tropism was also associated with lower CD4 + cell count, but not with higher HIV RNA levels. X4/DM tropic HIV strains were related to a higher frequency of transmitted reverse transcriptase resistance mutations in this unselected set of naive patients. As a consequence, if a patient harbors a non-CCR5 tropic virus and bears more reverse transcriptase resistance and less protease resistance, a boosted protease inhibitor-based first-line regimen should be preferred.
    Type of Medium: Online Resource
    ISSN: 1746-0794 , 1746-0808
    Language: English
    Publisher: Future Medicine Ltd
    Publication Date: 2013
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  • 2
    In: Pharmacogenomics, Future Medicine Ltd, Vol. 12, No. 4 ( 2011-04), p. 567-576
    Abstract: Aim: International HIV treatment guidelines recommend HLA-B*57:01 typing before abacavir administration, in order to reduce the incidence of abacavir hypersensitivity reactions, the major cause of early therapy discontinuation. A fast, sensitive and specific test for HLA-B*57:01 detection has been developed in the present study. Materials & methods: Two sets of sequence-specific primers were designed, and amplification rapidly detected by real-time PCR. Results: A total of 108 samples were analyzed in a single-blind fashion, and 41 samples were identified as positive. Complete agreement, with κ = 1 (standard error = 0.0962, p 〈 0.0001), was found, with a validated methodology used in the EPI109367 clinical trial funded by GlaxoSmithKline, and consisting of low-resolution sequence-specific oligonucleotide PCR, followed by high-resolution sequence-specific oligonucleotide PCR carried out on the HLA-B*57-positive samples. Conclusion: We provided a detailed characterization of a novel HLA-B*57:01 screening test, which can be easily implemented by those laboratories already involved in the detection of viral load and virus genotyping. Original submitted 26 October 2010; Revision submitted 13 December 2010.
    Type of Medium: Online Resource
    ISSN: 1462-2416 , 1744-8042
    Language: English
    Publisher: Future Medicine Ltd
    Publication Date: 2011
    SSG: 15,3
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  • 3
    In: Pharmacogenomics, Future Medicine Ltd, Vol. 15, No. 3 ( 2014-02), p. 319-327
    Abstract: Aim:HLA-B*57:01 status needs to be determined before initiating abacavir therapy. We developed a pharmacogenetic real-time (Q)-PCR screening test using two sets of sequence specific primers. This test has been implemented into routine clinical practice. Materials & methods: HIV-infected patients admitted at our University Hospital were thus genotyped using the above mentioned test. A panel of 80 DNA samples with a known genotype were used to characterize Q-PCR conditions using different master mixes. Results: A total of 353 patients were genotyped, detecting 15 (4.25%) HLA-B*57:01 positive carriers. Among the negative patients, 17.2% were treated with abacavir without any hypersensitivity reaction. Using different Q-PCR master mixes, significantly lower cutoff Ct values were found, thus new analytical settings are provided. Conclusion: The pharmacogenetic test developed in our laboratory for the fast screening of HLA-B*57:01 can be successfully implemented into routine clinical practice. All 16 sequences (including an additional six) currently known for the HLA-B*57:01 allele are detected by sequence specific primers used in this test. The Brilliant II SYBR ® Green QPCR MM (Stratagene) can safely replace the master mix originally used to develop the test. Original submitted 2 August 2013; Revision submitted 2 December 2013
    Type of Medium: Online Resource
    ISSN: 1462-2416 , 1744-8042
    Language: English
    Publisher: Future Medicine Ltd
    Publication Date: 2014
    SSG: 15,3
    Location Call Number Limitation Availability
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