Título:	Sensitivity of different DNA extraction methods and PCR to detect resistance in patients with leprosy stratified by the bacilloscopic index
Autor(es):	Sevilha-Santos, Lais Aquino, Danielle Costa Hans Neto, Günter Costa, Fabiano José Queiroz Sousa, Carlos Augusto Felipe de Morelo, Elaine Faria Santos Júnior, Agenor de Castro Moreira dos Gomes, Ciro Martins
ORCID:	https://orcid.org/0000-0003-2811-7537 https://orcid.org/0000-0001-8485-2603 https://orcid.org/0000-0002-3069-6884
Assunto:	Hanseníase Reação em cadeia da polimerase Baciloscopia Extração de DNA
Data de publicação:	2022
Editora:	Science Direct
Referência:	SEVILHA-SANTOS, Lais et al. Sensitivity of different DNA extraction methods and PCR to detect resistance in patients with leprosy stratified by the bacilloscopic index. The Brazilian Journal of Infectious Diseases, v. 26, n. 4, art. 102381, jul./ago. 2022. DOI 10.1016/j.bjid.2022.102381. Disponível em: https://www.sciencedirect.com/science/article/pii/S141386702200068X?via%3Dihub. Acesso em: 09 set. 2022.
Abstract:	Introduction: Antimicrobial resistance in leprosy is an emerging problem, and the quantitative impact of low bacilloscopic indexes (BIs) on the sensitivity of molecular tests is unknown. We aimed to evaluate the sensitivity of gene sequencing for the detection of mutations related to antimicrobial resistance in Mycobacterium leprae in patients with low BIs using an analytical model. Methods: Patients with leprosy were included and divided into two groups depending on their BIs (≥ 2+ and < 2+). The sensitivities of the two DNA extraction methods were compared after amplifying and sequencing the repetitive element (RLEP), folP1, rpoB and gyrA in M. leprae. Results: We included 56 patients with leprosy: 35 had BIs less than 2+ (22 had negative slitskin smear [SSS] results) and 21 patients had BIs greater than or equal to 2+. The sensitivity of the amplification of the RLEP target and the gene sequencing of folP1, rpoB and gyrA was 50 to 70% lower in patients with a BI less than 2+ and was significantly reduced in patients with lower BIs for all targets (p < 0.001). One patient had a mutation in the folP1 gene, and 14 patients had mutations in the gyrA gene, but no mutations related to antimicrobial resistance were found. Conclusions: We can conclude that the sensitivity of molecular tests is directly related to the BI, but these tests can still detect up to 20% of the targets in patients with BIs < 2+. New strategies to improve the sensitivity for detecting antimicrobial resistance in leprosy patients and reasonable clinical criteria for follow-up and the introduction of alternative treatments must be developed.
Licença:	The Brazilian Journal of Infectious Diseases - This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). Fonte: https://www.sciencedirect.com/science/article/pii/S141386702200068X?via%3Dihub. Acesso em: 09 set. 2022.
DOI:	https://doi.org/10.1016/j.bjid.2022.102381
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