Impact of qPCR on the diagnostic evaluation of individuals with suspected leprosy

Authors

DOI:

https://doi.org/10.33448/rsd-v12i3.40549

Keywords:

Leprosy; qPCR; Early diagnosis.

Abstract

Introduction: Leprosy is a chronic infection caused by Mycobacterium leprae, primarily affecting the peripheral nerves. It is known that despite the reduction in the number of new cases, transmission remains active due to the incidence of cases in children under 15 years of age. Thus, the search for more sensitive and specific diagnostic methods for the detection of subclinical infection becomes relevant. Objective: to verify, through qPCR, the presence of M. leprae in intradermal scrapings (IS) of individuals treated at CREDEN-PES/GV/MG. Method: Intradermal scraping (IS) samples were collected from 411 participants to determine the bacilloscopic index (BI), extraction, and amplification of M. leprae DNA. Participants were divided into two groups: GROUP 1, “Notified cases,” registered at SINAN; GROUP 2, “Individuals not notified,” who had a suspected diagnosis of leprosy. Of the total of 411 samples collected, 158 were obtained from notified cases and 253 from not notified individuals. Results: It was found that of the total number of cases, 58.86% (male) and 41.14% (female). Among the not notified individuals, 53.36% (male) and 46.64% (female). Considering the age range, both groups were aged (40 to 69 years). As for the IS, 34.18% (cases) and 0% (not notified) had a positive IS. In qPCR, 82.28% of cases and 47.43% of not notified individuals tested positive, with an illness odds ratio of 5.14. Conclusion: It is possible to infer those individuals with suspected leprosy, with positive qPCR, may be carriers of subclinical infection and potential transmitters of M. leprae to more susceptible individuals.

Author Biographies

Daisy Cristina Monteiro dos Santos, Universidade Federal de Juiz de Fora

Faculdade de Farmácia

Departamento Ciências Básicas da Vida

 

Heloine Martins Leite, Universidade Federal de Juiz de Fora

Programa Multicêntrico em Bioquimica e Biologia Molecular-PMBqBM

Lucia Alves de Oliveira Fraga, Universidade Federal de Juiz de Fora

Programa Multicêntrico em Bioquimica e Biologia Molecular-PMBqBM

Departamento de Ciências Básicas da Vida

References

Amorim, F. M., Nobre, M. L., Ferreira, L. C., Nascimento, L. S., Miranda, A. M., Monteiro, G. R. G., Dupnik, K. M., Duthie, M. S., Reed, S. G., & Jeronimo, S. M. B. (2016). Identifying leprosy and those at risk of developing leprosy by detection of antibodies against LID-1 and LID-NDO. PLoS Neglected Tropical Diseases, 10(9), e0004934. https://doi.org/10.1371/journal.pntd.0004934.

Araujo, S., Freitas, L. O., Goulart, L. R., & Goulart, I. M. B. (2016). Molecular evidence for the aerial route of infection ofMycobacterium lepraeand the role of asymptomatic carriers in the persistence of leprosy. Clinical Infectious Diseases: An Official Publication of the Infectious Diseases Society of America, 63(11), 1412–1420. https://doi.org/10.1093/cid/ciw570.

Bakker, M. I., Hatta, M., Kwenang, A., Van Mosseveld, P., Faber, W. R., Klatser, P. R., & Oskam, L. (2006). Risk factors for developing leprosy--a population-based cohort study in Indonesia. Leprosy Review, 77(1), 48–61. https://doi.org/10.47276/lr.77.1.48.

Banerjee, S., Sarkar, K., Gupta, S., Mahapatra, P. S., Gupta, S., Guha, S., Bandhopadhayay, D., Ghosal, C., Paine, S. K., Dutta, R. N., Biswas, N., & Bhattacharya, B. (2010). Multiplex PCR technique could be an alternative approach for early detection of leprosy among close contacts--a pilot study from India. BMC Infectious Diseases, 10(1), 252. https://doi.org/10.1186/1471-2334-10-252.

Brasil. (2010). Guia de procedimentos técnicos Baciloscopia em Hanseníase; Brasília: Ministério da Saúde. Ministério da Saúde. Secretaria de Vigilância em Saúde. Departamento de Vigilância Epidemiológica.

Brasil, (2021). Ministério da Saúde. NOTA TÉCNICA No 16/2021-CGDE/DCCI/SVS/MS. Recuperado 20 de fevereiro de 2023, de https://www.conass.org.br/wp-content/uploads/2021/07/SEI_MS-0020845770-Nota-Te%CC%81cnica-16.pdf

Cabral, P. B. e., Júnior, J. E. C., Macedo, A. C. de, Alves, A. R., Gonçalves, T. B., Cabral, T. C. B. e., Gondim, A. P. S., Pinto, M. I. M., Oseki, K. T., Camara, L. M. C., Rabenhorst, S. H. B., & Nagao-Dias, A. T. (2013). Anti-PGL1 salivary IgA/IgM, serum IgG/IgM, and nasal Mycobacterium leprae DNA in individuals with household contact with leprosy. International Journal of Infectious Diseases: IJID: Official Publication of the International Society for Infectious Diseases, 17(11), e1005–e1010. https://doi.org/10.1016/j.ijid.2013.05.011.

Caleffi, K. R., Hirata, R. D. C., Hirata, M. H., Caleffi, E. R., Siqueira, V. L. D., & Cardoso, R. F. (2012). Use of the polymerase chain reaction to detect Mycobacterium leprae in urine. Brazilian Journal of Medical and Biological Research, 45(2), 153–157. https://doi.org/10.1590/s0100-879x2012007500011.

Duthie, M. S., Goto, W., Ireton, G. C., Reece, S. T., Cardoso, L. P. V., Martelli, C. M. T., Stefani, M. M. A., Nakatani, M., de Jesus, R. C., Netto, E. M., Balagon, M. V. F., Tan, E., Gelber, R. H., Maeda, Y., Makino, M., Hoft, D., & Reed, S. G. (2007). Use of protein antigens for early serological diagnosis of leprosy. Clinical and Vaccine Immunology: CVI, 14(11), 1400–1408. https://doi.org/10.1128/cvi.00299-07.

Frade, M. A. C., de Paula, N. A., Gomes, C. M., Vernal, S., Bernardes Filho, F., Lugão, H. B., de Abreu, M. M. M., Botini, P., Duthie, M. S., Spencer, J. S., Soares, R. C. F. R., & Foss, N. T. (2017). Unexpectedly high leprosy seroprevalence detected using a random surveillance strategy in midwestern Brazil: A comparison of ELISA and a rapid diagnostic test. PLoS Neglected Tropical Diseases, 11(2), e0005375. https://doi.org/10.1371/journal.pntd.0005375.

Fiocruz. (2021). Fiocruz cria teste molecular para hanseníase inédito no Brasil. Recuperado 20 de fevereiro de 2023, de https://www.unasus.gov.br/noticia/fiocruz-cria-teste-molecular-para-hanseniase-inedito-no-brasil.

Gama, R. S., Souza, M. L. M. de, Sarno, E. N., Moraes, M. O. de, Gonçalves, A., Stefani, M. M. A., Garcia, R. M. G., & Fraga, L. A. de O. (2019). A novel integrated molecular and serological analysis method to predict new cases of leprosy amongst household contacts. PLoS Neglected Tropical Diseases, 13(6), e0007400. https://doi.org/10.1371/journal.pntd.0007400.

Goulart, I. M. B., & Goulart, L. R. (2008). Leprosy: diagnostic and control challenges for a worldwide disease. Archives of Dermatological Research, 300(6), 269–290. https://doi.org/10.1007/s00403-008-0857-y

Kang, T.-J., Kim, S.-K., Lee, S.-B., Chae, G.-T., & Kim, J.-P. (2003). Comparison of two different PCR amplification products (the 18-kDa protein gene vs. RLEP repetitive sequence) in the diagnosis of Mycobacterium leprae: PCR for diagnosis of M. leprae. Clinical and Experimental Dermatology, 28(4), 420–424. https://doi.org/10.1046/j.1365-2230.2003.01300.x.

Leite, M. L. C., Miranda, M. C. R., Pugedo, A. C., Grossi, M. A. F., Ramalho, K. C., & Costa, J. V. (2009). Situação Epidemiológica da Hanseníase Minas Gerais – Municípios Silenciosos – 2009-2018; 10° Simpósio Brasileiro de Hansenologia.

Martinez, A. N., Britto, C. F. P. C., Nery, J. A. C., Sampaio, E. P., Jardim, M. R., Sarno, E. N., & Moraes, M. O. (2006). Evaluation of real-time and conventional PCR targeting complex 85 genes for detection of Mycobacterium leprae DNA in skin biopsy samples from patients diagnosed with leprosy. Journal of Clinical Microbiology, 44(9), 3154–3159. https://doi.org/10.1128/jcm.02250-05.

Martinez, A. N., Lahiri, R., Pittman, T. L., Scollard, D., Truman, R., Moraes, M. O., & Williams, D. L. (2009). Molecular determination of Mycobacterium leprae viability by use of real-time PCR. Journal of Clinical Microbiology, 47(7), 2124–2130. https://doi.org/10.1128/jcm.00512-09.

Minas Gerais (2020). Plano Estadual de Saúde 2020-2023; Belo Horizonte: Secretaria de Estado de Saúde De Minas Gerais. Recuperado 20 de fevereiro de 2023, de https://www.saude.mg.gov.br/images/1_noticias/09_2021/01_jan-fev-marc/08-02-Plano-Estadual-de-Saude%20-de-Minas-Gerais%20%202020-2023.pdf.

Niitsuma, E. N. A., Bueno, I. de C., Arantes, E. O., Carvalho, A. P. M., Xavier Junior, G. F., Fernandes, G. da R., & Lana, F. C. F. (2021). Fatores associados ao adoecimento por hanseníase em contatos: revisão sistemática e metanálise. Revista brasileira de epidemiologia [Brazilian journal of epidemiology], 24. https://doi.org/10.1590/1980-549720210039.

Pêgoa, F., Procópioj, P. M., & Condév, A. S. ([s.d.]). Hanseníase: correlação entre o número de lesões hansênicas, nervos afetados e o diagnóstico precoce no estado de Minas Gerais. Revista Eletrônica Acervo Saúde, v, 12(9).

Pereira A. S. et. al. (2018). Metodologia da pesquisa científica. UFSM. https://www.ufsm.br/app/uploads/sites/358/2019/02/Metodologia-da-Pesquisa-Cientifica_final.pdf.

Reis, E. M., Araujo, S., Lobato, J., Neves, A. F., Costa, A. V., Gonçalves, M. A., Goulart, L. R., & Goulart, I. M. B. (2014). Mycobacterium leprae DNA in peripheral blood may indicate a bacilli migration route and high-risk for leprosy onset. Clinical Microbiology and Infection: The Official Publication of the European Society of Clinical Microbiology and Infectious Diseases, 20(5), 447–452. https://doi.org/10.1111/1469-0691.12349

Ribeiro, G. de C., Barreto, J. G., Bueno, I. de C., Vasconcelos, B. F., & Lana, F. C. F. (2019). Prevalence and spatial distribution of Mycobacterium leprae infection in a medium endemicity municipality. Rev Rene, 20, e39497. https://doi.org/10.15253/2175-6783.20192039497

Rocha, M. C. N., Nobre, M. L., & Garcia, L. P. (2020). Características epidemiológicas da hanseníase nos idosos e comparação com outros grupos etários, Brasil (2016-2018). Cadernos de saude publica, 36(9). https://doi.org/10.1590/0102/311x00048019

Sales, A. M., Ponce de Leon, A., Düppre, N. C., Hacker, M. A., Nery, J. A. C., Sarno, E. N., & Penna, M. L. F. (2011). Leprosy among patient contacts: A multilevel study of risk factors. PLoS Neglected Tropical Diseases, 5(3), e1013. https://doi.org/10.1371/journal.pntd.0001013

Scollard, D. M., Adams, L. B., Gillis, T. P., Krahenbuhl, J. L., Truman, R. W., & Williams, D. L. (2006). The continuing challenges of leprosy. Clinical Microbiology Reviews, 19(2), 338–381. https://doi.org/10.1128/cmr.19.2.338-381.2006

Worobec, S. (2012). Current approaches and future directions in the treatment of leprosy. Research and Reports in Tropical Medicine, 79. https://doi.org/10.2147/rrtm.s27395

Published

02/03/2023

How to Cite

PINHEIRO, M. D. S. .; OLIVEIRA, N. G. de .; SANTOS, D. C. M. dos .; LEITE, H. M. .; BAPTISTA, I. M. F. D. .; FAIRLEY, J. K. .; FRAGA, L. A. de O. . Impact of qPCR on the diagnostic evaluation of individuals with suspected leprosy. Research, Society and Development, [S. l.], v. 12, n. 3, p. e13312340549, 2023. DOI: 10.33448/rsd-v12i3.40549. Disponível em: https://rsdjournal.org/index.php/rsd/article/view/40549. Acesso em: 12 nov. 2024.

Issue

Section

Health Sciences