NDLI: Detección de mutaciones en rpoB relacionadas a rifampicina-resistencia en aislamientos clínicos de Mycobacterium tuberculosis a través de un ensayo simple y de bajo costo basado en High Resolution Melting Analysis

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Detección de mutaciones en rpoB relacionadas a rifampicina-resistencia en aislamientos clínicos de Mycobacterium tuberculosis a través de un ensayo simple y de bajo costo basado en High Resolution Melting Analysis

Content Provider	Semantic Scholar
Author	Calderón, Daniela Alvarez
Copyright Year	2019
Abstract	Tuberculosis is the leading cause of death from infectious disease. Particularly, Peru is in the top 30 for high MDR-TB burden countries. A proper and prompt drug susceptibility diagnosis is demanding to increase the chance for cure. Nowadays, phenotypic drug susceptibility test based on culture is still the reference test, though turnaround time is too prolonged. Molecular tests such as Genotype MTBDRplus and GeneXpert MTB/RIF are a quicker alternative for diagnosis but lack of easy-access for general population. In this study, we evaluated the efficiency of a versatile test based on high resolution-melting (HRM) assay for the detection of mutations in rpoB gene in rifampicin resistant M. tuberculosis clinical isolates, as a marker for multidrugresistance. DNA samples extracted with High Pure PCR Template Preparation kit from 123 clinical isolates, were included in the HRM analysis resulting in an overall sensitivity and specificity of 82.4%, 96.4%, respectively, employing DNA sequencing the reference standard test. Additionally, in order to reduce costs, DNA was extracted by phenol-chloroform method from sputum samples, encountering similar sensitivity values but increasing the HRM curves scattering. Our study also suggests a quantitative analysis for melting curves, using maximum relative fluorescence unit (RFUmax) values to set up a cut-off. Considering both types of analysis, we concluded that HRM is a robust, simple, rapid and low-cost test for the detection of rpoB mutations associated to rifampicin resistance.
File Format	PDF HTM / HTML
Alternate Webpage(s)	http://repositorio.upch.edu.pe/bitstream/handle/upch/6591/Deteccion_AlvarezCalderon_Daniela.pdf?isAllowed=y&sequence=1
Language	English
Access Restriction	Open
Content Type	Text
Resource Type	Article

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