Genetic Identification and Drug-Resistance Characterization of Mycobacterium tuberculosis Using a Portable Sequencing Device. A Pilot Study.
Jorge L CervantesNoemí Kaoru YokoboriBo-Young HongPublished in: Antibiotics (Basel, Switzerland) (2020)
Clinical management of tuberculosis (TB) in endemic areas is often challenged by a lack of resources including laboratories for Mycobacterium tuberculosis (Mtb) culture. Traditional phenotypic drug susceptibility testing for Mtb is costly and time consuming, while PCR-based methods are limited to selected target loci. We herein utilized a portable, USB-powered, long-read sequencing instrument (MinION), to investigate Mtb genomic DNA from clinical isolates to determine the presence of anti-TB drug-resistance conferring mutations. Data analysis platform EPI2ME and antibiotic-resistance analysis using the real time ARMA workflow, identified Mtb species as well as extensive resistance gene profiles. The approach was highly sensitive, being able to detect almost all described drug resistance conferring mutations based on previous whole genome sequencing analysis. Our findings are supportive of the practical use of this system as a suitable method for the detection of antimicrobial resistance genes, and effective in providing Mtb genomic information. Future improvements in the error rate through statistical analysis, drug resistance prediction algorithms and reference databases would make this a platform suited for the clinical setting. The small size, relatively inexpensive cost of the device, as well as its rapid and simple library preparation protocol and analysis, make it an attractive option for settings with limited laboratory infrastructure.
Keyphrases
- mycobacterium tuberculosis
- pulmonary tuberculosis
- data analysis
- genome wide
- antimicrobial resistance
- copy number
- randomized controlled trial
- high throughput
- machine learning
- emergency department
- single molecule
- loop mediated isothermal amplification
- healthcare
- dna methylation
- current status
- genome wide identification
- molecularly imprinted
- circulating tumor
- low cost
- electronic health record
- big data
- genome wide analysis
- transcription factor
- fluorescent probe
- high resolution
- genome wide association study
- living cells