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Related Concept Videos

Mutations01:39

Mutations

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Mutations01:35

Mutations

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Mutations are changes in the sequence of DNA. These changes can occur spontaneously or they can be induced by exposure to environmental factors. Mutations can be characterized in a number of different ways: whether and how they alter the amino acid sequence of the protein, whether they occur over a small or large area of DNA, and whether they occur in somatic cells or germline cells.
Chromosomal Alterations Are Large-Scale Mutations
While point mutations are changes in a single nucleotide in...
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Viral Mutations00:36

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A mutation is a change in the sequence of bases of DNA or RNA in a genome. Some mutations occur during replication of the genome due to errors made by the polymerase enzymes that replicate DNA or RNA. Unlike DNA polymerase, RNA polymerase is prone to errors because it is not capable of “proofreading” its work. Viruses with RNA-based genomes, like HIV, therefore accrue mutations faster than viruses with DNA-based genomes. Because mutation and recombination provide the raw material...
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In a population that is not at Hardy-Weinberg equilibrium, the frequency of alleles changes over time. Therefore, any deviations from the five conditions of Hardy-Weinberg equilibrium can alter the genetic variation of a given population. Conditions that change the genetic variability of a population include mutations, natural selection, non-random mating, gene flow, and genetic drift (small population size).
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Mutations in Microorganisms01:18

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Mutations are heritable changes in an organism’s genome involving alterations in the base sequence of DNA or RNA. These changes can influence cellular processes and phenotypic traits, potentially transforming the unaltered wild type into a mutant form. Such changes, termed forward mutations, are pivotal in shaping the genetic diversity of organisms.RNA viruses exhibit the highest mutation rates due to the absence of robust proofreading mechanisms during genome replication. In contrast,...
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Point mutations are genetic alterations involving the change of a single nucleotide base pair in DNA. Depending on how the alteration affects protein synthesis, they can lead to various consequences.Point mutations fall into the following types:Silent mutations occur when a nucleotide change does not alter the amino acid sequence due to the redundancy of the genetic code. For instance, changing ACC to ACA still encodes threonine, leaving the protein function unaffected. This occurs because...
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Detection of Rare Mutations in CtDNA Using Next Generation Sequencing
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Comprehensive antibiotic-linked mutation assessment by resistance mutation sequencing (RM-seq).

Romain Guérillot1, Lucy Li1, Sarah Baines1

  • 1Department of Microbiology and Immunology, The University of Melbourne at the Doherty Institute for Infection & Immunity, Melbourne, Victoria, Australia.

Genome Medicine
|September 1, 2018
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Summary

This study introduces RM-seq, a novel deep sequencing method for detecting bacterial antibiotic resistance mutations. RM-seq enables high-throughput analysis of resistance in mixed populations, aiding in surveillance and characterization.

Keywords:
Antibiotic resistanceDaptomycinDeep sequencingMycobacterium tuberculosisResistance mutationsRifampicinStaphylococcus aureus

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Area of Science:

  • Microbiology
  • Genomics
  • Molecular Biology

Background:

  • Bacterial antibiotic resistance is a growing global health threat.
  • Mutation acquisition is a key, yet poorly understood, driver of resistance.
  • Current methods struggle to detect low-frequency resistance mutations in complex populations.

Purpose of the Study:

  • To develop a high-throughput sequencing workflow for detecting and assessing bacterial antibiotic resistance mutations.
  • To enable the characterization of resistance mechanisms in mixed bacterial populations.
  • To facilitate the study of rare resistant subpopulations and their associated phenotypes.

Main Methods:

  • Development of RM-seq, an amplicon-based deep sequencing workflow.
  • Adaptation of a molecular barcoding technique from Low Error Amplicon sequencing (LEA-seq).
  • Application of RM-seq to detect and quantify resistance mutations in vitro and in vivo.

Main Results:

  • RM-seq enables sensitive detection of very low-frequency resistant sub-populations.
  • The workflow allows for high-throughput functional assessment of mutational resistance.
  • Accurate quantification of resistance mutations facilitates phenotypic screening (e.g., persistence, cross-resistance).

Conclusions:

  • RM-seq provides a powerful tool for comprehensive detection, characterization, and surveillance of resistant bacterial populations.
  • This method can advance our understanding of antibiotic resistance evolution and spread.
  • RM-seq aids in identifying rare resistant strains during infections and characterizing their resistance profiles.