Experimental platform utilising melting curve technology for detection of mutations in Mycobacterium tuberculosis isolates

Autor: Nicki Casali, James Hume, Agnieszka Broda, Richard Bowker, Gemma Blackwell, Francis Drobniewski, Huma Khan, Waqar Hussain, Vlad Nikolayevskyy, Bhakti Patel
Přispěvatelé: Innovate UK, Imperial College Trust
Rok vydání: 2018
Předmět:
0301 basic medicine
Antitubercular Agents
SUSCEPTIBILITY
Nucleic Acid Denaturation
0302 clinical medicine
Drug Resistance
Multiple
Bacterial
Drugs resistance
Tuberculosis
Multidrug-Resistant
030212 general & internal medicine
11 Medical and Health Sciences
Melting curves
biology
Isoniazid
Molecular assays
General Medicine
3. Good health
Infectious Diseases
Original Article
medicine.symptom
Rifampin
Life Sciences & Biomedicine
medicine.drug
Microbiology (medical)
Tuberculosis
Point-of-Care Systems
030106 microbiology
Microbial Sensitivity Tests
DIAGNOSIS
Rapid detection
Microbiology
Sensitivity and Specificity
Melting curve analysis
Mycobacterium tuberculosis
03 medical and health sciences
Multidrug-resistant tuberculosis
medicine
Humans
RIFAMPIN RESISTANCE
Tuberculosis
Pulmonary
METAANALYSIS
Science & Technology
Base Sequence
business.industry
Sputum
RAPID DETECTION
Sequence Analysis
DNA
06 Biological Sciences
medicine.disease
biology.organism_classification
Virology
Mutation
Nontuberculous mycobacteria
business
Rifampicin
Zdroj: European Journal of Clinical Microbiology & Infectious Diseases
ISSN: 1435-4373
Popis: Tuberculosis (TB) remains one of the most deadly infections with approximately a quarter of cases not being identified and/or treated mainly due to a lack of resources. Rapid detection of TB or drug-resistant TB enables timely adequate treatment and is a cornerstone of effective TB management. We evaluated the analytical performance of a single-tube assay for multidrug-resistant TB (MDR-TB) on an experimental platform utilising RT-PCR and melting curve analysis that could potentially be operated as a point-of-care (PoC) test in resource-constrained settings with a high burden of TB. Firstly, we developed and evaluated the prototype MDR-TB assay using specimens extracted from well-characterised TB isolates with a variety of distinct rifampicin and isoniazid resistance conferring mutations and nontuberculous Mycobacteria (NTM) strains. Secondly, we validated the experimental platform using 98 clinical sputum samples from pulmonary TB patients collected in high MDR-TB settings. The sensitivity of the platform for TB detection in clinical specimens was 75% for smear-negative and 92.6% for smear-positive sputum samples. The sensitivity of detection for rifampicin and isoniazid resistance was 88.9 and 96.0% and specificity was 87.5 and 100%, respectively. Observed limitations in sensitivity and specificity could be resolved by adjusting the sample preparation methodology and melting curve recognition algorithm. Overall technology could be considered a promising PoC methodology especially in resource-constrained settings based on its combined accuracy, convenience, simplicity, speed, and cost characteristics. Electronic supplementary material The online version of this article (10.1007/s10096-018-3246-2) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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