ArticleMicrobiologyOpen2019
Three-reaction high-resolution melting assay for rapid differentiation of Mycobacterium tuberculosis complex members.
Article in MicrobiologyOpen, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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9 citing papers in PubMed.
- Evidence ofFrontiers in veterinary science · 2025Article
- Dynamic Challenges of Active Tuberculosis: Prevalence and Risk Factors of Co-Infection in Clinical and Microbiological Characteristics at King Abdulaziz University Hospital, Jeddah, Saudi Arabia.International journal of general medicine · 2025Article
- Culture-independent detection ofFrontiers in veterinary science · 2025Article
- Insights into mycobacteriome composition in Mycobacterium bovis-infected African buffalo (Syncerus caffer) tissue samples.Scientific reports · 2024Article
- Identification and molecular characterization ofOne health (Amsterdam, Netherlands) · 2024Article
- Whole genome sequencing-based identification of human tuberculosis caused by animal-lineageJournal of clinical microbiology · 2023Article
- Single-nucleotide polymorphism-based epidemiological analysis of KoreanJournal of veterinary science · 2021Article
- Article
- Three-reaction high-resolution melting assay for rapid differentiation of Mycobacterium tuberculosis complex members.MicrobiologyOpen · 2019Article
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4 authors.
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Abstract
The possibility of introducing a reliable assay for a quick identification and differentiation of the main species of Mycobacterium tuberculosis complex (MTBC) supports the improvement of efficient tuberculosis combating strategies worldwide. Commercially available assays are often based on cultured samples; however, due to the long cultivation time of mycobacteria, results are delayed. Developed PCR approaches have been published previously, though, when testing intricate veterinary samples, the complex composition of multiplex qPCRs frequently leads to assay failure. In order to overcome those limits, a paradigm of a three-reaction high-resolution melting (HRM) assay for the simultaneous identification and differentiation of the main members of MTBC was established. The assay is based on single nucleotide polymorphisms within gyrB and gyrA, which have been used as target for the establishment of two highly specific HRM assays (HRM assays 1 and 2) discriminating M. tuberculosis/ Mycobacterium canetti, Mycobacterium bovis/M. bovis BCG, Mycobacterium caprae/rare M. caprae/M. bovis ecotypes, Mycobacterium africanum/Mycobacterium orygis/ Mycobacterium pinnipedii/Clade A1, Mycobacterium microti, and a rare subtype of M. canettii followed by a third HRM assay (HRM assay 3) allowing a further differentiation of M. bovis, M. bovis BCG, and a rare subtype of M. caprae/M. bovis, which is considered to be a novel ecotype. High-resolution melting assay 1 is described in a previously published report. High-resolution melting assay 2 showed 100% correlation of all 39 examined isolates with the results of a commercial identification kit. 96% of the clinical samples tested demonstrated concordant results. High-resolution melting assay 3 showed an accordance of 100% with the results of the commercially available identification kit of all 22 samples analyzed. The proposed strategy of the three-reaction HRM assay can be used for an accurate differentiation of up to seven groups of MTBC and potentially to identify a rare subtype of M. canettii either on isolates or on clinical samples.
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