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 |
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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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