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Basic Characteristics of Mutations
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Mutation Site
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Y181C |
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Mutation Site Sentence
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Integrated WT 001-005 contained the wild-type codon, and Integrated DRM 001-005 contained DRM-conferring nucleotide substitutions for K65R, K103N, V106M, Y181C, M184VI, and G190A (Supplementary Table 9). |
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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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RT |
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Standardized Encoding Gene
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gag-pol:155348
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Genotype/Subtype
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HIV-1 |
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Viral Reference
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HXB2
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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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NNRTIs |
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Location
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Los Alamos |
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Literature Information
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PMID
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33603155
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Title
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PANDAA intentionally violates conventional qPCR design to enable durable, mismatch-agnostic detection of highly polymorphic pathogens
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Author
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MacLeod IJ,Rowley CF,Essex M
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Journal
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Communications biology
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Journal Info
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2021 Feb 18;4(1):227
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Abstract
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Sensitive and reproducible diagnostics are fundamental to containing the spread of existing and emerging pathogens. Despite the reliance of clinical virology on qPCR, technical challenges persist that compromise their reliability for sustainable epidemic containment as sequence instability in probe-binding regions produces false-negative results. We systematically violated canonical qPCR design principles to develop a Pan-Degenerate Amplification and Adaptation (PANDAA), a point mutation assay that mitigates the impact of sequence variation on probe-based qPCR performance. Using HIV-1 as a model system, we optimized and validated PANDAA to detect HIV drug resistance mutations (DRMs). Ultra-degenerate primers with 3' termini overlapping the probe-binding site adapt the target through site-directed mutagenesis during qPCR to replace DRM-proximal sequence variation. PANDAA-quantified DRMs present at frequency >/=5% (2 h from nucleic acid to result) with a sensitivity and specificity of 96.9% and 97.5%, respectively. PANDAA is an innovative advancement with applicability to any pathogen where target-proximal genetic variability hinders diagnostic development.
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Sequence Data
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-
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