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Basic Characteristics of Mutations
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Mutation Site
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H69Y |
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Mutation Site Sentence
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Table 1. AVA® and SmartGene® analyses |
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Mutation Level
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Amino acid level |
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Mutation Type
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Nonsynonymous substitution |
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Gene/Protein/Region
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PR |
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Standardized Encoding Gene
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gag-pol
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Genotype/Subtype
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HIV-1 |
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Viral Reference
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K03455
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Functional Impact and Mechanisms
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Disease
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HIV Infections
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Immune
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- |
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Target Gene
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-
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Clinical and Epidemiological Correlations
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Clinical Information
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- |
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Treatment
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PIs |
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Location
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France |
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Literature Information
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PMID
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29856864
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Title
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Evaluation of different analysis pipelines for the detection of HIV-1 minority resistant variants
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Author
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Perrier M,Desire N,Storto A,Todesco E,Rodriguez C,Bertine M,Le Hingrat Q,Visseaux B,Calvez V,Descamps D,Marcelin AG,Charpentier C
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Journal
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PloS one
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Journal Info
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2018 Jun 1;13(6):e0198334
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Abstract
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OBJECTIVE: Reliable detection of HIV minority resistant variants (MRVs) requires bioinformatics analysis with specific algorithms to obtain good quality alignments. The aim of this study was to analyze ultra-deep sequencing (UDS) data using different analysis pipelines. METHODS: HIV-1 protease, reverse transcriptase (RT) and integrase sequences from antiretroviral-naive patients were obtained using GS-Junior(R) (Roche) and MiSeq(R) (Illumina) platforms. MRVs were defined as variants harbouring resistance-mutation present at a frequency of 1%-20%. Reads were analyzed using different alignment algorithms: Amplicon Variant Analyzer(R), Geneious(R) compared to SmartGene(R) NGS HIV-1 module. RESULTS: 101 protease and 51 RT MRVs identified in 139 protease and 124 RT sequences generated with a GS-Junior(R) platform were analyzed using AVA(R) and SmartGene(R) software. The correlation coefficients for the MRVs were R2 = 0.974 for protease and R2 = 0.972 for RT. Discordances (n = 13 in protease and n = 15 in RT) mainly concerned low-level MRVs (i.e., with frequencies of 1%-2%, n = 18/28) and they were located in homopolymeric regions (n = 10/15). Geneious(R) and SmartGene(R) software were used to analyze 143 protease, 45 RT and 26 integrase MRVs identified in 172 protease, 69 RT, and 72 integrase sequences generated with a MiSeq(R) platform. The correlation coefficients for the MRVs were R2 = 0.987 for protease, R2 = 0.995 for RT and R2 = 0.993 for integrase. Discordances (n = 9 in protease, n = 3 in RT, and n = 3 in integrase) mainly concerned low-level MRVs (n = 13/15). CONCLUSION: We found an excellent correlation between the various UDS analysis pipelines that we tested. However, our results indicate that specific attention should be paid to low-level MRVs, for which the use of two different analysis pipelines and visual inspection of sequences alignments might be beneficial. Thus, our results argue for use of a 2% threshold for MRV detection, rather than the 1% threshold, to minimize misalignments and time-consuming sight reading steps essential to ensure accurate results for MRV frequencies below 2%.
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Sequence Data
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-
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