AI Article Synopsis

  • The study investigates the use of next-generation sequencing to identify genetic mutations related to antibiotic resistance in clinical samples, particularly focusing on the bacterium that is challenging to culture and requires molecular methods for detection.* -
  • In the analysis of 98 specimens, the researchers found mutations in 94 samples, totaling 469 single nucleotide polymorphisms (SNPs), which could help create comprehensive drug resistance profiles for macrolides and fluoroquinolones within a typical turnaround time of 3 days.* -
  • The proposed method promises to enhance the understanding and diagnosis of antibiotic resistance mechanisms, paving the way for its potential adoption in clinical laboratories after thorough validation.*

Article Abstract

is fastidious to culture, and its detection in human clinical specimens relies mainly on molecular methods. Phenotypic determination of antibiotic susceptibility for this bacterium is not a timely or feasible option for most clinical laboratories. This study sought to determine whether next-generation sequencing technologies can effectively be employed in determining genetic mutations associated with drug resistance in samples collected in Aptima Hologic tubes and possibly integrating them into viable workflows in public health laboratories. Following analysis by a custom-designed bioinformatics pipeline, at least one mutation/sample has been identified in 94/98 specimens in at least one of seven loci (macrolides: , , ; fluoroquinolones: , , , ) described previously to be connected to antibiotic resistance. This method identified a total of 469 single nucleotide polymorphisms (SNPs) (452 mutations): 134 of 23S rRNA SNPs and 318 amino acid mutations: 114 substitutions and 204 synonymous; the turnaround time (sample to analyzed sequence) was typically 3 days. The assays and workflows described in this work demonstrated that the determination of a drug resistance profile for macrolides and fluoroquinolones of samples by using next-generation sequencing in clinical samples is a feasible approach that can be implemented in clinical laboratories, following thorough and extensive validation studies.IMPORTANCEThe mechanisms of drug resistance in are complex and involve several genetic loci. The molecular methods for accurately characterizing resistance to fluoroquinolones and macrolides in this organism are often not available or approved for patient use and do not cover all genetic determinants. To this end, we propose a next-generation sequencing-based method with a turnaround time of 3 days that includes the investigation of all drug resistance loci of . Following adaptation, validation, and verification for routine clinical use, assays based on this method may yield molecular results that can be used to guide proper treatment regimens and for surveillance of drug resistance in the general population.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10913745PMC
http://dx.doi.org/10.1128/spectrum.03845-23DOI Listing

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