Examinando por Materia "Molecular Diagnostic Techniques"

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    DNA extraction from samples fixed on slides for molecular detection of Mycobacterium Leprae Lima, 2023
    (Universidad Privada Norbert Wiener, 2024-08-03) Vidia Vitelia, Cotrina Maccha; Huamán Cárdenas, Victor Raul
    Leprosy is an infectious disease caused by Mycobacterium leprae. In Peru, the standard laboratory diagnosis is bacilloscopy from smear-fixed samples. In recent years, other countries have started using PCR testing as an alternative for diagnosis due to its high sensitivity. Objective: To extract DNA from smear-fixed samples for molecular detection of Mycobacterium leprae. Methodology: The study included 50 samples from skin incision scrapings fixed on glass slides (30 samples with positive bacilloscopy and 20 samples with negative bacilloscopy). DNA was extracted using a silica column and the 16S rRNA sequence was amplified by real-time PCR (qPCR). Results: The quality of the extracted DNA ranged from low to good, which did not affect amplification through qPCR. DNA concentrations ranged from 0.5 ng/mL to 67.2 ng/mL. qPCR results showed 100% categorical agreement for the 30 positive samples and 40% disagreement for the 20 negative samples. Conclusion: Good results were obtained from DNA extraction of Mycobacterium leprae from smear-fixed skin incision scraping samples.