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      Sensitivity of different DNA extraction methods and PCR to detect resistance in patients with leprosy stratified by the bacilloscopic index

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          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 slit-skin 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.

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          Most cited references26

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          Leprosy: review of the epidemiological, clinical, and etiopathogenic aspects - Part 1*

          Leprosy is caused by Mycobacterium leprae and has been known since biblical times. It is still endemic in many regions of the world and a public health problem in Brazil. The prevalence rate in 2011 reached 1.54 cases per 10,000 inhabitants in Brazil. The mechanism of transmission of leprosy consists of prolonged close contact between susceptible and genetically predisposed individuals and untreated multibacillary patients. Transmission occurs through inhalation of bacilli present in upper airway secretion. The nasal mucosa is the main entry or exit route of M. leprae. The deeper understanding of the structural and biological characteristics of M. leprae, the sequencing of its genome, along with the advances in understanding the mechanisms of host immune response against the bacilli, dependent on genetic susceptibility, have contributed to the understanding of the pathogenesis, variations in the clinical characteristics, and progression of the disease. This article aims to update dermatologist on epidemiological, clinical, and etiopathogenic leprosy aspects.
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            Multidrug resistant Mycobacterium leprae from patients with leprosy.

            Sequences of the folP1, rpoB, and gyrA genes were analyzed for 88 isolates of Mycobacterium leprae from leprosy patients in Japan, Haiti, Indonesia, Pakistan, and the Philippines. Thirteen isolates (14.8%) showed representative mutations in more than two genes, suggesting the emergence of multidrug-resistant M. leprae.
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              Diagnostic accuracy of tests for leprosy: a systematic review and meta-analysis

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                Author and article information

                Contributors
                Journal
                Braz J Infect Dis
                Braz J Infect Dis
                The Brazilian Journal of Infectious Diseases
                Elsevier
                1413-8670
                1678-4391
                27 June 2022
                Jul-Aug 2022
                27 June 2022
                : 26
                : 4
                : 102381
                Affiliations
                [a ]Faculdade de Medicina, Programa de Pós-graduação em Ciências Médicas, Universidade de Brasília, Brasília, DF, Brazil
                [b ]Universidade de Brasília, Hospital Universitário de Brasília, Brasília, DF, Brazil
                [c ]Laboratório Central de Saúde Pública do Distrito Federal, Secretaria de Saúde do Distrito Federal, Brasília, DF, Brazil
                [d ]Núcleo de Medicina Tropical, Universidade de Brasília, Brasília, DF, Brazil
                Author notes
                [* ]Corresponding author at: Faculdade de Medicina, Programa de Pós-graduação em Ciências Médicas, Universidade de Brasília, Brasília, DF, Brazil. cirogomes@ 123456unb.br
                Article
                S1413-8670(22)00068-X 102381
                10.1016/j.bjid.2022.102381
                9459058
                35772498
                fefc70d6-c3fe-43f8-91b9-0be7d5308b86
                © 2022 Sociedade Brasileira de Infectologia. Published by Elsevier España, S.L.U.

                This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

                History
                : 10 March 2022
                : 7 June 2022
                Categories
                Original Article

                leprosy,microbial sensitivity tests,polymerase chain reaction,therapeutics,bacilloscopic index,dna extraction

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