Urine-based ELISA using a recombinant chimeric protein for the diagnosis of paucibacillary and multibacillary leprosy - 26/02/25

Doi : 10.1016/j.tube.2025.102613

Raquel S.B. Câmara ^a,^{^{1
These authors contributed equally for the study.}}, Isabela A.G. Pereira ^a,^{^{1
These authors contributed equally for the study.}}, Geise C. Espíndola ^a,^{^{1
These authors contributed equally for the study.}}, Daniela P. Lage ^a, Ana L. Silva ^a, Camila S. Freitas ^a, Bárbara P.N. Assis ^a,^b, Laís V.A. Corrêa ^a, Ricardo L.F. Moreira ^b, Sandra Lyon ^b, Rozana C. Silva ^b, Tiago S. Barros ^b, Ana Laura G. de Oliveira ^a, Fernanda Ludolf ^a,^c, Miguel A. Chávez-Fumagalli ^d, Myron Christodoulides ^e, Ricardo A. Machado-de-Ávila ^f, Unaí Tupinambás ^a, Denise U. Gonçalves ^a, Manoel O. da Costa Rocha ^a, Eduardo A.F. Coelho ^a,^g,^⁎,^{^{1
These authors contributed equally for the study.}} , Ana T. Chaves ^a,^{^{1
These authors contributed equally for the study.}}
^a Programa de Pós-Graduação em Ciências da Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Belo Horizonte, 30130-100, Minas Gerais, Brazil
^b Fundação Hospitalar do Estado de Minas Gerais, Hospital Eduardo de Menezes, Belo Horizonte, 30622-020, Minas Gerais, Brazil
^c Programa de Pós-Graduação em Ciências da Saúde, Faculdade de Ciências Médicas de Minas Gerais, Belo Horizonte, 30130-110, Minas Gerais, Brazil
^d Computational Biology and Chemistry Research Group, Vicerrectorado de Investigación, Universidad Católica de Santa María, Urb. San José S/N, Umacollo, Arequipa, 04000, Peru
^e Neisseria Research Group, Molecular Microbiology, School of Clinical and Experimental Sciences, University of Southampton Faculty of Medicine, Southampton General Hospital, Southampton, England, UK
^f Programa de Pós-Graduação em Ciências da Saúde, Universidade do Extremo Sul Catarinense, Criciúma, 88806-000, Santa Catarina, Brazil
^g Departamento de Patologia Clínica, COLTEC, Universidade Federal de Minas Gerais, Av. Antônio Carlos, 6627, Belo Horizonte, Minas Gerais, Brazil

^⁎Corresponding author.Laboratório de Pesquisa do Programa de Pós-Graduação em Ciências da Saúde: Infectologia e Medicina Tropical, Faculdade de Medicina, Universidade Federal de Minas Gerais, Avenida Prof. Alfredo Balena, 190, 30130-100, Belo Horizonte, Minas Gerais, Brazil.Laboratório de Pesquisa do Programa de Pós-Graduação em Ciências da Saúde: Infectologia e Medicina TropicalFaculdade de MedicinaUniversidade Federal de Minas GeraisAvenida Prof. Alfredo Balena190Belo HorizonteMinas Gerais30130-100Brazil

Abstract

Leprosy diagnosis is difficult to perform due to variable sensitivity and/or specificity of the tests. In addition, the collection of the blood samples requires laboratorial structure and trained professionals. In the present study, the diagnostic efficacy of M1 chimeric protein, which was recently showed to be antigenic for leprosy using a serum-based ELISA, was evaluated against patient urine. Paired serum and urine samples were collected from patients with paucibacillary (PB) and multibacillary (MB) leprosy, tegumentary and visceral leishmaniasis, tuberculosis, Chagas disease, malaria, and HIV-infected subjects. Samples from healthy individuals and household contacts were also used. The protein and peptides used to compose it were used as antigens, and results showed that the four peptides presented good sensitivity and specificity to detect MB leprosy, while M1 protein showed sensitivity and specificity of 98.5 % and 100 %, respectively, to detect both PB and MB leprosy, when an urine-based ELISA was performed. Positive (PPV) and negative (NPV) predictive values were 100 % and 98.3 %, respectively. In a serum-based ELISA, sensitivity and specificity were 96.9 % and 100 %, respectively, with PPV and NPV of 100 % and 96.5 %, respectively. In conclusion, preliminary data suggest that M1 protein could be considered for diagnosis of leprosy by using patient urine.

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Highlights

•	M1 chimeric protein was tested against urine and serum samples of leprosy patients.
•	The antigen was used in ELISA experiments against 310 paired samples.
•	It showed high sensitivity and specificity values to diagnose the disease.
•	Both pauci- and multibacillary clinic forms were diagnosed by using M1 antigen.
•	Urine can be considered as an alternative biological analyte to diagnose leprosy.

Le texte complet de cet article est disponible en PDF.

Keywords : Leprosy, Chimeric protein, Diagnosis, ELISA, Urine, Serum

Plan

Introduction

Materials and methods

Production of the recombinant M1 protein and synthetic peptides

Ethics

Study population

Diagnosis of diseases

Reactivity of the synthetic peptides against patient urine samples

Serological evaluation using M1 protein for diagnosis of leprosy

Urine-based ELISA using M1 protein for diagnosis of leprosy

Discussion

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Urine-based ELISA using a recombinant chimeric protein for the diagnosis of paucibacillary and multibacillary leprosy - 26/02/25

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