RESUMO
BACKGROUND: The impact of nutrient availability on the survival of Mycobacterium leprae and the development of leprosy remains largely unknown. Iron is essential for the survival and replication of pathogens, while vitamin D has been involved with pathogen elimination and immunoregulation. OBJECTIVES: We evaluated the influence of dietary iron and vitamin D supplementation and restriction on the inflammatory response of mouse immune cells in vitro. METHODS: After 30 days of standard or modified diets, peritoneal cells and splenocytes were stimulated with the alive microorganisms and sonicated antigens of M. leprae, respectively. The production of inflammatory cytokines, reactive oxygen species, and cell proliferation were evaluated. FINDINGS: In peritoneal cells, vitamin D supplementation and iron restriction reduced the production of IL-6 and TNF in response to M. leprae, while splenocytes presented a reduction in TNF production under the same conditions. Lower levels of IFN-γ and TNF were observed in both iron-supplemented and iron-deficient splenocytes. Besides, iron supplementation also reduced the production of IL-6 and IL-10. No changes in the production of reactive oxygen species or in cell proliferation were observed related to different diets. MAIN CONCLUSIONS: Taken together, these data point to an interference of the status of these nutrients on the interaction between the host and M. leprae, with the potential to interfere with the progression of leprosy. Our results highlight the impact of nutritional aspects on this neglected disease, which is significantly associated with unfavourable social conditions.
Assuntos
Citocinas , Mycobacterium leprae , Espécies Reativas de Oxigênio , Baço , Vitamina D , Animais , Mycobacterium leprae/imunologia , Mycobacterium leprae/efeitos dos fármacos , Vitamina D/farmacologia , Vitamina D/administração & dosagem , Baço/imunologia , Camundongos , Citocinas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Ferro/metabolismo , Proliferação de Células/efeitos dos fármacos , Suplementos Nutricionais , Feminino , Hanseníase/imunologia , Masculino , Camundongos Endogâmicos BALB CRESUMO
Leprosy continues to pose a significant challenge to public health, particularly in certain global regions. Accurate diagnosis and understanding of the disease's etiology are crucial for effective management and prevention. This study aimed to explore the contribution of Natural resistance-associated macrophage protein 1 (NRAMP1) and its genetic variations, as well as the levels of anti-PGL-1 antibodies, to the pathology of multibacillary leprosy in affected individuals and their household contacts. The study included 23 multibacillary leprosy patients and 28 household contacts. NRAMP1 protein expression and anti-PGL-1 IgG and IgM levels were measured using PCR and ELISA techniques, respectively. Genotypic variants of the NRAMP1 gene were also examined. Statistical analyses, including Mann-Whitney tests and univariate logistic regression, were employed to evaluate the data. Significant differences were observed in NRAMP1 protein expression and IgG and IgM levels between the patient and household contact groups. The study also highlighted the role of the NRAMP1 gene and its D543N and 3'UTR polymorphisms in leprosy susceptibility. No significant differences were observed in the genotype variants of INT4 between the two groups. These findings emphasize the potential of integrating PCR technology with serological tests to enhance diagnostic precision in leprosy. They also suggest the need for further research to clarify the role of NRAMP1 and its polymorphisms in leprosy susceptibility and resistance.
Assuntos
Anticorpos Antibacterianos , Antígenos de Bactérias , Proteínas de Transporte de Cátions , Glicolipídeos , Imunoglobulina M , Hanseníase Multibacilar , Humanos , Masculino , Hanseníase Multibacilar/genética , Feminino , Adulto , Imunoglobulina M/sangue , Anticorpos Antibacterianos/sangue , Proteínas de Transporte de Cátions/genética , Pessoa de Meia-Idade , Antígenos de Bactérias/imunologia , Antígenos de Bactérias/genética , Imunoglobulina G/sangue , Adulto Jovem , Genótipo , Adolescente , Características da Família , Expressão Gênica , Mycobacterium leprae/imunologia , Mycobacterium leprae/genéticaRESUMO
BACKGROUND: Leprosy is a chronic infectious disease caused by Mycobacterium leprae (M. leprae) that is responsible for deformities and irreversible peripheral nerve damage and has a broad spectrum of clinical and serological manifestations. Leprosy primarily affects the peripheral nerves and rarely presents with central nervous system involvement. Diagnosing leprosy can still be difficult in some cases, especially when the infection involves uncommon clinical manifestations and extracutaneous sites. Delayed diagnosis and treatment of leprosy may lead to irreversible damage and death. CASE PRESENTATION: We report a case of a 30-year-old female presenting with "repeated high fever with symptoms of headache for 14 days". On the day of admission, physical signs of lost eyebrows and scattered red induration patches all over her body were observed. The patient's diagnosis was based on the clinical characteristics using a combination of metagenomic next-generation sequencing (mNGS) of cerebrospinal fluid (CSF) and slit-skin smear. After confirming Listeria meningitis and multibacillary leprosy with erythema nodosum leprosum (ENL), a type 2 reaction, she was treated with ampicillin sodium, dapsone, rifampicin, clofazimine, methylprednisolone, and thalidomide. At the 1-year follow-up, the frequency and severity of headaches have significantly decreased and a good clinical response with improved skin lesions was found. CONCLUSION: This case highlights the importance of considering leprosy, which is a rare and underrecognized disease, in the differential diagnosis of skin rashes with rheumatic manifestations, even in areas where the disease is not endemic, and physicians should be alerted about the possibility of central nervous system infections. In addition, mNGS can be used as a complementary diagnostic tool to traditional diagnostic methods to enhance the diagnostic accuracy of leprosy.
Assuntos
Sequenciamento de Nucleotídeos em Larga Escala , Mycobacterium leprae , Humanos , Feminino , Adulto , Mycobacterium leprae/genética , Mycobacterium leprae/isolamento & purificação , Mycobacterium leprae/efeitos dos fármacos , Hanseníase/diagnóstico , Hanseníase/líquido cefalorraquidiano , Hanseníase/microbiologia , Hanseníase/tratamento farmacológico , Metagenômica , Líquido Cefalorraquidiano/microbiologia , Hansenostáticos/uso terapêuticoRESUMO
BACKGROUND: In December 2023, our hospital confirmed a case of systemic lupus erythematosus complicated with Mycobacterium leprae infection. The patient has extensive patchy erythema on the back and face, with obvious itching. There are multiple subcutaneous masses on both hands, some of which are accompanied by tenderness, wave sensation, and other symptoms. The patient's mother has a history of leprosy and close contact with the patient. The patient tested positive for syphilis antibodies 2 years ago and did not receive formal treatment. There is no other history of chronic illness. METHODS: Under local anesthesia, the left hand skin lesion was excised, followed by tissue pathological biopsy, acid-fast staining, mNGS, and serum Treponema pallidum antibody detection. RESULTS: Pathological biopsy results: A large number of foam-like histiocytes, lymphocytes, and plasma cells were mainly found in the superficial and deep layers of the dermis, as well as around the blood vessels and sweat glands in the subcutaneous fat. Cellulose-like degeneration is seen in some blood vessel walls. Tissue acid-fast staining: positive, tissue mNGS detection: Mycobacterium leprae. CLINICAL DIAGNOSIS: 1. Borderline leprosy, 2. Subacute cutaneous lupus erythematosus. Treat with methylprednisolone 32 mg qd po + aluminum magnesium suspension 15 mL tid po + calcium carbonate D3 tablets 0.6 g qd po + rifampicin 450 mg qd po + dapsone 100 mg qd. After 10 days of treatment, the patient improved and was discharged from the hospital. CONCLUSIONS: Mycobacterium leprae infection occurs during SLE treatment and is often difficult to distinguish from skin symptoms caused by SLE. In the clinical treatment of infectious diseases, the effect of conventional anti-bacterial drugs is not good. The auxiliary examination indicates severe infection and the routine culture is negative. The possibility of special pathogen infection should be considered in combination with the medical history. With the popularity of new detection methods such as mNGS, the importance of traditional smear detection methods cannot be ignored.
Assuntos
Lúpus Eritematoso Sistêmico , Mycobacterium leprae , Humanos , Mycobacterium leprae/isolamento & purificação , Lúpus Eritematoso Sistêmico/complicações , Lúpus Eritematoso Sistêmico/diagnóstico , Feminino , Hanseníase/diagnóstico , Hanseníase/microbiologia , Hanseníase/tratamento farmacológico , Hanseníase/complicações , AdultoRESUMO
The assessment of ELISA plates coated with phenolic glycolipid-I/PGL-I revealed excellent stability during eight years of storage at room temperature, promoting consistent IgM antibody detection in multibacillary leprosy patients. These stable, standardized plates can significantly contribute to efficient leprosy serology research and support its widespread distribution and use in endemic countries.
Assuntos
Anticorpos Antibacterianos , Antígenos de Bactérias , Ensaio de Imunoadsorção Enzimática , Glicolipídeos , Imunoglobulina M , Mycobacterium leprae , Imunoglobulina M/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Mycobacterium leprae/imunologia , Humanos , Anticorpos Antibacterianos/imunologia , Glicolipídeos/imunologia , Antígenos de Bactérias/imunologia , Hanseníase/diagnóstico , Hanseníase/microbiologia , Hanseníase/imunologia , Fatores de TempoRESUMO
OBJECTIVE: To compare the diagnostic efficacy of multiplex polymerase chain reaction (PCR), Mycobacterium leprae-specific repetitive element (RLEP) PCR and loop-mediated isothermal amplification (LAMP) PCR in the diagnosis of pediatric leprosy as an alternative to slit-skin smear (SSS) examination. METHODS: A cross-sectional study was performed on 26 children aged 0-18 years with characteristic skin lesions of leprosy. SSS examination for acid fast bacilli (AFB) was performed for all children. Additionally, urine, stool and blood samples were tested by three PCR techniques - multiplex, RLEP and LAMP. The results of these tests were compared with each other and with results of SSS examination for acid fast bacilli (AFB) using appropriate statistical tests. RESULTS: Out of 26 patients studied, SSS examination was positive for AFB in 7 cases (26.9%). In blood samples, the positivity of multiplex PCR, RLEP PCR and LAMP PCR was 84.6%, 80.8%, and 80.8%, respectively. Multiplex PCR in blood samples was positive in 100% (n = 7) of SSS positive cases and 84.2% (16 out of 19) of the SSS negative cases (P < 0.001). The positivity of all PCR methods in urine and stool samples was significantly lesser than in blood. CONCLUSION: Multiplex PCR in blood sample is a superior diagnostic tool for pediatric leprosy compared to RLEP PCR and LAMP PCR as well as SSS examination.
Assuntos
Fezes , Hanseníase , Reação em Cadeia da Polimerase Multiplex , Humanos , Criança , Hanseníase/diagnóstico , Estudos Transversais , Pré-Escolar , Adolescente , Lactente , Reação em Cadeia da Polimerase Multiplex/métodos , Masculino , Feminino , Fezes/microbiologia , Técnicas de Amplificação de Ácido Nucleico/métodos , Técnicas de Amplificação de Ácido Nucleico/normas , Mycobacterium leprae/isolamento & purificação , Mycobacterium leprae/genética , Reação em Cadeia da Polimerase/métodos , Reação em Cadeia da Polimerase/normas , Recém-Nascido , Sensibilidade e Especificidade , Técnicas de Diagnóstico MolecularRESUMO
Leprosy is a chronic infectious disease caused by the bacillus Mycobacterium leprae. The disease may evolve for inflammatory reactions, reversal reaction (RR) and erythema nodosum leprosum (ENL), the major cause of irreversible neuropathy in leprosy, which occur in 1 in 3 people with leprosy, even with effective treatment of M. leprae. Leprosy remains persistently endemic in our region where it predominantly affects lowest socioeconomic conditions people, as Toxoplasma gondii infection in the municipality studied. Previously, we have shown T. gondii coinfection as a risk marker for leprosy, mainly in its severe form. This present study assessed whether T. gondii infection is also a risk factor for leprosy reactions and the predictive value of immunoglobulin production prior to development of leprosy reactions. Patients with leprosy (n = 180), co-infected or not with T. gondii, had their serum investigated for levels of IgA, IgE, IgG1, IgG2, IgG3 and IgG4 anti-PGL-1 by ELISA prior to development of leprosy reactions. The serologic prevalence for T. gondii infection was 87.7% in leprosy reaction patients reaching 90.9% in those with ENL. The leprosy reaction risk increased in T. gondii seropositive individuals was two-fold ([OR] = 2.366; 95% confidence interval [CI 95%]: 1.024-5.469) higher than those seronegative, and considering the risk of ENL, this increase was even more evident (OR = 6.753; 95% CI: 1.050-72.85) in coinfected individuals. When evaluated the prediction of anti-PGL-1 immunoglobulin levels for development of leprosy reactions in patients coinfected or not with T. gondii, only the increase IgE levels were associated to occurrence of reactional episodes of leprosy, specifically ENL type, in patients coinfected with T. gondii, compared to those not coinfected or no reaction. Thus, the immunomodulation in co-parasitism T. gondii-M. leprae suggest increased levels of IgE as a biomarker for early detection of these acute inflammatory episodes and thereby help prevent permanent neuropathy and disability in leprosy patients.
Assuntos
Eritema Nodoso , Imunoglobulina E , Toxoplasma , Toxoplasmose , Humanos , Toxoplasmose/sangue , Toxoplasmose/complicações , Toxoplasmose/imunologia , Toxoplasmose/epidemiologia , Eritema Nodoso/imunologia , Eritema Nodoso/epidemiologia , Eritema Nodoso/sangue , Feminino , Masculino , Adulto , Imunoglobulina E/sangue , Pessoa de Meia-Idade , Toxoplasma/imunologia , Coinfecção/imunologia , Coinfecção/parasitologia , Mycobacterium leprae/imunologia , Adulto Jovem , Adolescente , Fatores de Risco , Idoso , Hanseníase Virchowiana/imunologia , Hanseníase Virchowiana/complicações , Hanseníase Virchowiana/sangue , Hanseníase Virchowiana/epidemiologiaRESUMO
Leprosy, caused by the bacterium Mycobacterium leprae, is known to primarily affect the skin and peripheral nerves. We present a rare case of leprosy initially manifesting as demyelinating polyneuropathy. A 46-year-old female presented with progressive weakness, tingling, and numbness in her extremities. Nerve conduction studies revealed evidence of demyelination, prompting further investigations. Skin slit-skin smears confirmed the diagnosis of leprosy, with the presence of acid-fast bacilli. The patient was subsequently started on multidrug therapy, leading to significant clinical improvement. This case highlights the importance of considering leprosy as a differential diagnosis in patients presenting with demyelinating polyneuropathy, especially in endemic regions.
Assuntos
Hanseníase , Mycobacterium leprae , Polirradiculoneuropatia Desmielinizante Inflamatória Crônica , Humanos , Feminino , Pessoa de Meia-Idade , Polirradiculoneuropatia Desmielinizante Inflamatória Crônica/diagnóstico , Polirradiculoneuropatia Desmielinizante Inflamatória Crônica/tratamento farmacológico , Hanseníase/diagnóstico , Hanseníase/tratamento farmacológico , Hanseníase/microbiologia , Hanseníase/complicações , Diagnóstico Diferencial , Mycobacterium leprae/isolamento & purificação , Mycobacterium leprae/genética , Pele/patologia , Pele/microbiologia , Hansenostáticos/uso terapêuticoRESUMO
Leprosy is a chronic granulomatous infectious and disabling disease caused by two mycobacteria, Mycobacterium leprae and Mycobacterium lepromatosis. Acute inflammatory responses, known as leprosy reactions, are significant contributors to disabilities. Three types of leprosy reactions have been identified based on excessive cytokine release (e.g. type 1) or the accumulation of immune complexes in tissues inducing multiorgan damage (e.g. types 2 and 3). The type of leprosy reaction has implications on treatment and management strategies, yet are not well understood by health workers caring for leprosy patients. We attempt to describe the immunologic mechanisms behind the different leprosy reactions and the rationale for tailoring clinical treatment and management to the particular type of leprosy reaction based on the underlying immunologic situation.
Assuntos
Hanseníase , Mycobacterium leprae , Humanos , Hanseníase/imunologia , Hanseníase/microbiologia , Hanseníase/patologia , Mycobacterium leprae/imunologia , Citocinas/metabolismoRESUMO
BACKGROUND: Post-exposure prophylaxis (PEP) using single-dose rifampicin reduces progression from infection with Mycobacterium leprae to leprosy disease. We compared effectiveness of different administration modalities, using a higher (20 mg/kg) dose of rifampicin-single double-dose rifampicin (SDDR)-PEP. METHODS: We did a cluster randomised study in 16 villages in Madagascar and 48 villages in Comoros. Villages were randomly assigned to four study arms and inhabitants were screened once a year for leprosy, for 4 consecutive years. All permanent residents (no age restriction) were eligible to participate and all identified patients with leprosy were treated with multidrug therapy (SDDR-PEP was provided to asymptomatic contacts aged ≥2 years). Arm 1 was the comparator arm, in which no PEP was provided. In arm 2, SDDR-PEP was provided to household contacts of patients with leprosy, whereas arm 3 extended SDDR-PEP to anyone living within 100 m. In arm 4, SDDR-PEP was offered to household contacts and to anyone living within 100 m and testing positive to anti-phenolic glycolipid-I. The main outcome was the incidence rate ratio (IRR) of leprosy between the comparator arm and each of the intervention arms. We also assessed the individual protective effect of SDDR-PEP and explored spatial associations. This trial is registered with ClinicalTrials.gov, NCT03662022, and is completed. FINDINGS: Between Jan 11, 2019, and Jan 16, 2023, we enrolled 109â436 individuals, of whom 95â762 had evaluable follow-up data. Our primary analysis showed a non-significant reduction in leprosy incidence in arm 2 (IRR 0·95), arm 3 (IRR 0·80), and arm 4 (IRR 0·58). After controlling for baseline prevalence, the reduction in arm 3 became stronger and significant (IRR 0·56, p=0·0030). At an individual level SDDR-PEP was also protective with an IRR of 0·55 (p=0·0050). Risk of leprosy was two to four times higher for those living within 75 m of an index patient at baseline. INTERPRETATION: SDDR-PEP appears to protect against leprosy but less than anticipated. Strong spatial associations were observed within 75 m of index patients. Targeted door-to-door screening around index patients complemented by a blanket SDDR-PEP approach will probably have a substantial effect on transmission. FUNDING: European and Developing Countries Clinical Trials Partnership. TRANSLATION: For the French translation of the abstract see Supplementary Materials section.
Assuntos
Hansenostáticos , Hanseníase , Profilaxia Pós-Exposição , Rifampina , Humanos , Hanseníase/prevenção & controle , Hanseníase/tratamento farmacológico , Hanseníase/epidemiologia , Masculino , Feminino , Adulto , Rifampina/administração & dosagem , Rifampina/uso terapêutico , Hansenostáticos/uso terapêutico , Hansenostáticos/administração & dosagem , Profilaxia Pós-Exposição/métodos , Pessoa de Meia-Idade , Adolescente , Adulto Jovem , Madagáscar/epidemiologia , Criança , Análise por Conglomerados , Incidência , Mycobacterium lepraeRESUMO
Leprosy, one of the oldest recorded diseases in human history, remains prevalent in Asia, Africa, and South America, with over 200,000 cases every year.1,2 Although ancient DNA (aDNA) approaches on the major causative agent, Mycobacterium leprae, have elucidated the disease's evolutionary history,3,4,5 the role of animal hosts and interspecies transmission in the past remains unexplored. Research has uncovered relationships between medieval strains isolated from archaeological human remains and modern animal hosts such as the red squirrel in England.6,7 However, the time frame, distribution, and direction of transmissions remains unknown. Here, we studied 25 human and 12 squirrel samples from two archaeological sites in Winchester, a medieval English city well known for its leprosarium and connections to the fur trade. We reconstructed four medieval M. leprae genomes, including one from a red squirrel, at a 2.2-fold average coverage. Our analysis revealed a phylogenetic placement of all strains on branch 3 as well as a close relationship between the squirrel strain and one newly reconstructed medieval human strain. In particular, the medieval squirrel strain is more closely related to some medieval human strains from Winchester than to modern red squirrel strains from England, indicating a yet-undetected circulation of M. leprae in non-human hosts in the Middle Ages. Our study represents the first One Health approach for M. leprae in archaeology, which is centered around a medieval animal host strain, and highlights the future capability of such approaches to understand the disease's zoonotic past and current potential.
Assuntos
Genoma Bacteriano , Hanseníase , Mycobacterium leprae , Filogenia , Sciuridae , Animais , Mycobacterium leprae/genética , Mycobacterium leprae/isolamento & purificação , Sciuridae/microbiologia , Hanseníase/microbiologia , Hanseníase/história , Humanos , Inglaterra , DNA Antigo/análise , Arqueologia , História MedievalRESUMO
The diagnosis if leprosy is difficult, as it requires clinical expertise and sensitive laboratory tests. In this study, we develop a serological test for leprosy by using bioinformatics tools to identify specific B-cell epitopes from Mycobacterium leprae hypothetical proteins, which were used to construct a recombinant chimeric protein, M1. The synthetic peptides were obtained and showed good reactivity to detect leprosy patients, although the M1 chimera have showed sensitivity (Se) and specificity (Sp) values higher than 90.0% to diagnose both paucibacillary (PB) and multibacillary (MB) leprosy patients, but not those developing tegumentary or visceral leishmaniasis, tuberculosis, Chagas disease, malaria, histoplasmosis and aspergillosis, in ELISA experiments. Using sera from household contacts, values for Se and Sp were 100% and 65.3%, respectively. In conclusion, our proof-of-concept study has generated data that suggest that a new recombinant protein could be developed into a diagnostic antigen for leprosy.
Assuntos
Antígenos de Bactérias , Proteínas de Bactérias , Epitopos de Linfócito B , Hanseníase , Mycobacterium leprae , Sensibilidade e Especificidade , Humanos , Mycobacterium leprae/imunologia , Mycobacterium leprae/genética , Epitopos de Linfócito B/imunologia , Epitopos de Linfócito B/genética , Antígenos de Bactérias/imunologia , Antígenos de Bactérias/genética , Hanseníase/diagnóstico , Hanseníase/imunologia , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/genética , Ensaio de Imunoadsorção Enzimática/métodos , Adulto , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Masculino , Feminino , Testes Sorológicos/métodos , Biologia Computacional/métodos , Pessoa de Meia-Idade , Adulto Jovem , AdolescenteRESUMO
The nine-banded armadillo (Dasypus novemcinctus) is currently considered an invasive species in parts of its range in the USA, and this range continues to expand to the north and east. Nine-banded armadillos are one of a handful of mammals known to contract leprosy (also known as Hansen's disease); range expansion thus leads to public health concerns about whether this might increase human exposure to infected animals. We collected blood samples from 61 road-killed armadillos over two summers (2021 and 2022) in Tennessee, a US state near the northern extreme of the species' current range, and screened them for exposure to Mycobacterium leprae, the causative agent of leprosy. All animals were seronegative, providing no evidence that range expansion is increasing the distribution of leprosy in the US.
Assuntos
Tatus , Hanseníase , Mycobacterium leprae , Animais , Tatus/microbiologia , Hanseníase/veterinária , Hanseníase/epidemiologia , Tennessee/epidemiologia , Estudos Soroepidemiológicos , Mycobacterium leprae/imunologia , Feminino , MasculinoRESUMO
Despite low prevalence of leprosy worldwide, new cases continue to present and require swift evaluation and diagnosis to prevent complications. Here, we describe a case of lepromatous leprosy with Lucio's phenomenon initially presenting with facial and periorbital edema. A 38-year-old Brazilian woman presented to the emergency department with facial swelling and erythema, initially treated as cellulitis. Due to rapid worsening despite broad-spectrum antibiotics, she underwent soft tissue exploration and biopsy due to concern for necrotizing fasciitis. During her course, she also developed retiform purpura of bilateral upper and lower extremities. Periorbital and lower extremity pathological specimens ultimately revealed acid-fast bacilli consistent with Mycobacterium leprae , and the patient improved with multidrug therapy. This case illustrates the diagnostic difficulty of lepromatous leprosy with Lucio's phenomenon, which can initially present with periorbital edema.
Assuntos
Edema , Hanseníase Virchowiana , Humanos , Feminino , Hanseníase Virchowiana/diagnóstico , Hanseníase Virchowiana/tratamento farmacológico , Hanseníase Virchowiana/microbiologia , Adulto , Edema/diagnóstico , Edema/etiologia , Mycobacterium leprae/isolamento & purificação , Diagnóstico Diferencial , Infecções Oculares Bacterianas/diagnóstico , Infecções Oculares Bacterianas/microbiologia , Biópsia , Doenças Orbitárias/diagnóstico , Doenças Orbitárias/microbiologia , Hansenostáticos/uso terapêuticoRESUMO
The World Health Organization (WHO) aims to reduce new leprosy cases by 70% by 2030, necessitating advancements in leprosy diagnostics. Here we discuss the development of two WHO's target product profiles for such diagnostics. These profiles define criteria for product use, design, performance, configuration and distribution, with a focus on accessibility and affordability. The first target product profile outlines requirements for tests to confirm diagnosis of leprosy in individuals with clinical signs and symptoms, to guide multidrug treatment initiation. The second target product profile outlines requirements for tests to detect Mycobacterium leprae or M. lepromatosis infection among asymptomatic contacts of leprosy patients, aiding prophylactic interventions and prevention. Statistical modelling was used to assess sensitivity and specificity requirements for these diagnostic tests. The paper highlights challenges in achieving high specificity, given the varying endemicity of M. leprae, and identifying target analytes with robust performance across leprosy phenotypes. We conclude that diagnostics with appropriate product design and performance characteristics are crucial for early detection and preventive intervention, advocating for the transition from leprosy management to prevention.
L'Organisation mondiale de la Santé (OMS) vise à réduire le nombre de nouveaux cas de lèpre de 70% d'ici 2030, ce qui nécessite un meilleur diagnostic de la maladie. Dans le présent document, nous évoquons le développement de deux profils de produit cible établis par l'OMS à cette fin. Ces profils définissent des critères en matière d'utilisation, de conception, de performances, de configuration et de distribution du produit, en accordant une attention particulière à l'accessibilité et à l'abordabilité. Le premier profil de produit cible décrit les exigences pour les tests servant à confirmer le diagnostic de la lèpre chez les individus qui présentent des signes cliniques et des symptômes, afin d'orienter l'instauration d'un traitement à base de plusieurs médicaments. Le second profil de produit cible décrit les exigences pour les tests servant à détecter une infection à Mycobacterium leprae ou M. lepromatosis parmi les contacts asymptomatiques de patients lépreux, ce qui contribue à l'adoption de mesures prophylactiques et à la prévention. Nous avons eu recours à une modélisation statistique pour évaluer les exigences de sensibilité et de spécificité de ces tests diagnostiques. Cet article met en évidence les obstacles à l'atteinte d'un niveau élevé de spécificité en raison de l'endémicité variable de M. leprae, et à l'identification d'analytes cibles offrant de bons résultats chez les phénotypes lépreux. Nous concluons qu'un diagnostic reposant sur des caractéristiques de performance et de conception appropriées est essentiel pour détecter rapidement la maladie et intervenir en amont, et nous plaidons pour une prévention plutôt qu'une gestion de la lèpre.
La Organización Mundial de la Salud (OMS) pretende reducir los nuevos casos de lepra en un 70% para 2030, lo que requiere avances en el diagnóstico de la lepra. Aquí se analiza el desarrollo de dos perfiles de productos objetivo de la OMS para este tipo de diagnósticos. Estos perfiles definen los criterios de uso, diseño, rendimiento, configuración y distribución de los productos, centrándose en su accesibilidad y asequibilidad. El primer perfil de producto objetivo describe los requisitos de las pruebas para confirmar el diagnóstico de la lepra en personas con signos y síntomas clínicos, con el fin de orientar el inicio del tratamiento con múltiples fármacos. El segundo perfil de producto objetivo describe los requisitos de las pruebas para detectar la infección por Mycobacterium leprae o M. lepromatosis entre los contactos asintomáticos de los pacientes con lepra, para facilitar las intervenciones profilácticas y la prevención. Se utilizaron modelos estadísticos para evaluar los requisitos de sensibilidad y especificidad de estas pruebas diagnósticas. El artículo destaca las dificultades para lograr una alta especificidad, dada la diferente endemicidad de M. leprae, y para identificar analitos diana con un rendimiento sólido en todos los fenotipos de lepra. Concluimos que los diagnósticos con un diseño de producto y unas características de rendimiento adecuados son fundamentales para la detección precoz y la intervención preventiva, lo que favorece la transición del manejo de la lepra a la prevención.
Assuntos
Hanseníase , Humanos , Hanseníase/diagnóstico , Hanseníase/tratamento farmacológico , Mycobacterium leprae/genética , Sensibilidade e Especificidade , Modelos Estatísticos , Diagnóstico PrecoceRESUMO
ABSTRACT: Leprosy, an ancient disease, continues to be a public health concern as it remains endemic in several countries. After reaching the elimination target (1/10,000) as a public health problem in 2005 in India, around 1.2 lakh cases have been detected every year over the last decade indicating active transmission of leprosy bacillus (Mycobacterium leprae). Single-nucleotide polymorphisms (SNPs), genomic insertions/deletions and variable-number tandem repeats (VNTRs) have been identified as genetic markers for tracking M. leprae transmission. As the leprosy bacilli cannot be cultured in vitro, molecular testing of M. leprae genotypes is done by polymerase chain reaction-based sequencing which provides a practical alternative for the identification of strains as well as drug resistance-associated mutations. Whole-genome sequencing (WGS) of M. leprae directly from clinical samples has also proven to be an effective tool for identifying genetic variations which can further help refine the molecular epidemiological schemes based on SNPs and VNTRs. However, the WGS data of M. leprae strains from India are scarce, being responsible for a gross under-representation of the genetic diversity of M. leprae strains present in India and need to be addressed suitably. Molecular studies of leprosy can provide better insight into phylogeographic markers to monitor the transmission dynamics and emergence of antimicrobial resistance. An improved understanding of M. leprae transmission is essential to guide efficient leprosy control strategies. Therefore, this review compiles and discusses the current status of molecular epidemiology, genotyping and the potential of genome-wide analysis of M. leprae strains in the Indian context.
Assuntos
Hanseníase , Mycobacterium leprae , Humanos , DNA Bacteriano/genética , Hanseníase/epidemiologia , Hanseníase/genética , Epidemiologia Molecular , Mycobacterium leprae/genética , Polimorfismo de Nucleotídeo Único/genética , ÍndiaRESUMO
Leprosy diagnosis is difficult due to the clinical similarity with other infectious diseases, and laboratory tests presents problems related to sensitivity and/or specificity. In this study, we used bioinformatics to assess Mycobacterium leprae proteins and formulated a chimeric protein that was tested as a diagnostic marker for the disease. The amino acid sequences from ML0008, ML0126, ML0308, ML1057, ML2028, ML2038, ML2498 proteins were evaluated, and the B-cell epitopes QASVAYPATSYADFRAHNHWWNGP, SLQRSISPNSYNTARVDP and QLLGQTADVAGAAKSGPVQPMGDRGSVSPVGQ were considered M. leprae-specific and used to construct the gene encoding the recombinant antigen. The gene was constructed, the recombinant protein was expressed, purified and tested in ELISA using 252 sera, which contained samples from multibacillary (MB) or paucibacillary (PB) leprosy patients, from their household contacts and healthy individuals, as well as from patients with Chagas disease, visceral and tegumentary leishmaniases (VL/TL), malaria, tuberculosis, and HIV. Sensitivity (Se) and specificity (Sp) for MB and PB samples compared to sera from both healthy subjects and individuals with cross-reactive diseases were 100%. The Se value for MB and PB samples compared to sera from household contacts was 100%, but Sp was 64%. In conclusion, data suggest that this protein could be considered in future studies for leprosy diagnosis.
Assuntos
Antígenos de Bactérias , Proteínas de Bactérias , Ensaio de Imunoadsorção Enzimática , Epitopos de Linfócito B , Hanseníase Multibacilar , Hanseníase Paucibacilar , Mycobacterium leprae , Testes Sorológicos , Mycobacterium leprae/imunologia , Mycobacterium leprae/genética , Humanos , Epitopos de Linfócito B/imunologia , Testes Sorológicos/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Antígenos de Bactérias/imunologia , Antígenos de Bactérias/genética , Hanseníase Paucibacilar/diagnóstico , Hanseníase Paucibacilar/imunologia , Proteínas de Bactérias/imunologia , Proteínas de Bactérias/genética , Hanseníase Multibacilar/diagnóstico , Hanseníase Multibacilar/imunologia , Anticorpos Antibacterianos/sangue , Proteínas Recombinantes de Fusão/imunologia , Valor Preditivo dos Testes , Feminino , Masculino , Sensibilidade e Especificidade , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/genéticaRESUMO
BACKGROUND: Mycobacterium leprae causes leprosy that is highly stigmatized and chronic infectious skin disease. Only some diagnostic tools are being used for the identification M. leprae in clinical samples, such as bacillary detection, and histopathological tests. These methods are invasive and often have low sensitivity. Currently, the PCR technique has been used as an effective tool fordetecting M. leprae DNA across different clinical samples. The current study aims to detect M. leprae DNA in urine samples of untreated and treated leprosy patients using the Rlep gene (129 bp) and compared the detection among Ridley-Jopling Classification. METHODS: Clinical samples (Blood, Urine, and Slit Skin Smears (SSS)) were collected from leprosy and Non-leprosy patients. DNA extraction was performed using standard laboratory protocol and Conventional PCR was carried out for all samples using Rlep gene target and the amplicons of urine samples were sequenced by Sanger sequencing to confirm the Rlep gene target. RESULTS: The M. leprae DNA was successfully detected in all clinical samples across all types of leprosy among all the study groups using RLEP-PCR. Rlep gene target was able to detect the presence of M. leprae DNA in 79.17% of urine, 58.33% of blood, and 50% of SSS samples of untreated Smear-Negative leprosy patients. The statistical significant difference (p = 0.004) was observed between BI Negative (Slit Skin Smear test) and RLEP PCR positivity in urine samples of untreated leprosy group. CONCLUSION: The PCR positivity using Rlep gene target (129 bp) was highest in all clinical samples among the study groups, across all types of leprosy. Untreated tuberculoid and PNL leprosy patients showed the highest PCR positivity in urine samples, indicating its potential as a non-invasive diagnostic tool for leprosy and even for contact screening.