Multibacillary Leprosy Patients Research Articles

A recombinant chimeric antigen constructed with B-cell epitopes from Mycobacterium leprae hypothetical proteins is effective for the diagnosis of leprosy

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.

Serodiagnosis of paucibacillary and multibacillary leprosy using a recombinant chimeric protein composed of specific B-cell epitopes derived from Mycobacterium leprae proteins

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.

Plantar pressure distribution and altered postural control in multibacillary leprosy patients

BackgroundLeprosy is a chronic infectious disease caused by Mycobacterium leprae, predominantly affecting the peripheral nerves, resulting in sensory and motor deficits in the feet. Foot ulcers and imbalances are frequent manifestations in leprosy, often correlating with diminished sensitivity. While clinical scales and monofilament esthesiometers are conventionally utilized to evaluate foot sensitivity and balance in these patients, their discriminatory power is limited and their effectiveness is greatly dependent on the examiner’s proficiency. In contrast, baropodometry and posturography offer a more comprehensive evaluation, aiming to preempt potential damage events. This study aimed was to assess the correlation between baropodometry and force plate measurements in leprosy patients and control participants, to improve the prevention and treatment of foot ulcers and complications associated with leprosy.MethodologyThis cross-sectional study was conducted during 2022 and enrolled 39 participants (22 patients with multibacillary leprosy and 17 non-leprosy controls). Demographic data were collected, and a monofilament esthesiometer was used to assess sensory deficits. In addition, physical examinations and balance and plantar pressure tests were conducted.The Student’s t-test was used to compare mean and maximum plantar pressures between groups. For most COP variables, a Mann-Whitney Wilcoxon test was used, except for AP amplitude which was analyzed with the Student’s t-test due to its normal distribution. The relationship between foot pressure and balance control was assessed using Spearman’s correlation, focusing on areas with significant pressure differences between groups.Principal findingsLeprosy patients showed increased pressure in forefoot areas (T1, M1, T2-T5, and M2) and decreased pressure in hindfoot regions (MH and LH) compared to controls. These patients also displayed higher AP and ML amplitudes, suggesting poorer COP control. Correlation analyses between the two groups revealed that foot plantar pressures significantly impact balance control. Specifically, increased T1 region pressures correlated with greater sway in balance tasks, while decreased MH region pressures were linked to reduced COP control.Conclusions/significanceThe findings suggest a joint disturbance of plantar pressure distribution and static balance control in leprosy patients. These alterations may increase the risk of tissue injuries, including calluses and deformities, as well as falls.

Levels of TGF- Serum Positively Correlated with Levels of IgM Anti PGL-1 In Household Contacts of Multibacillary Leprosy Patients

Background Leprosy is a chronic progressive infectious disease. Phenolic glycolipid-1 (PGL-1) is an antigen of Mycobacterium leprae, which can trigger the host’s antibody response. Transforming growth factorβ (TGF-β) plays an immunosuppressive role when the host is exposed to PGL-1 antigen or other M. leprae antigens. This study aims to determine the correlation of TGF-β levels with IgM anti-PGL-1 levels in patients with multibacillary leprosy household contacts. Methods Observational analytical study with a cross-sectional approach. The study subjects consisted of 48 household contact subjects and 24 non-household contact subjects aged 15-65 years old who were selected by consecutive sampling based on inclusion and exclusion criteria. 3 mL of venous blood samples were taken and then examined for IgM anti-PGL-1 and TGF-β with the ELISA kit. Data analysis was carried out using SPSS version 23, and a p-value <0.05 was significant. Results The mean level of IgM anti-PGL-1 in the household contact group was 685.46 ± 290.79 u/mL, while in the non-household contact group was 345.50 ± 206.58 u/mL. The mean TGF-β level in household contact groups was 256.69 ± 127.41 pg/mL, while in the non-household contact group was 144.85 ± 36.73 pg/mL (p<0.001). This study found a moderate positive relationship (r=0.450, p<0.001) between levels of TGF-β and IgM anti-PGL-1 household contacts and non-household contacts group. Conclusions The mean level of IgM anti-PGL-1 and TGF-β in household contacts is higher than in non-household contacts, with a significant difference. There is a moderate positive significant relationship between levels of TGF-β and IgM anti-PGL-1 household contacts and non-household contacts group.

Comparison between lymecycline with multidrug therapy and standard multidrug regimen (WHO-MDT) in the treatment of multibacillary leprosy patients: a retrospective cohort study.

Hansen's disease or leprosy is a chronic, infectious disease that has locally and globally afflicted all populations. Despite standard treatment with multidrug therapy (WHO-MDT), the incidence of drug resistance has been an increasingly prevalent global problem in leprosy management. This study compared the effectiveness between lymecycline with WHO-MDT and standard WHO-MDT in leprosy treatment. The research is a retrospective cohort study at a tertiary hospital from January 2011 to July 2021. Pre- and post-treatment bacillary index, presence of new lesions, nerve function impairment, and leprosy reactions were obtained through chart review. The results showed a significant difference in bacteriological index (BI) in both groups at the end of the treatment. However, a higher reduction in BI was noted for the lymecycline group. For the group that took WHO-MDT alone, BI decreased by 0.7 (P < 0.001) whereas patients who took lymecycline and WHO-MDT had a BI difference of 3 (P < 0.001) upon completion of treatment. A significant decrease in the recurrence of lesions (P = 0.006) and nerve function impairment (P = 0.038) was also noted in the lymecycline group whereas there was no significant difference in leprosy reactions between the two groups. Lymecycline 600 mg daily for 3 months can be used as an adjunct in cases of leprosy resistance and treatment failure among multibacillary patients. Lymecycline significantly reduced bacillary index, recurrence of skin lesions, and nerve function impairment through its possible immunomodulatory, antiapoptotic, and neuroprotective effects.

New cases of childhood leprosy in two male siblings in low endemic province: an iceberg phenomenon?

Children are the leprosy population most at risk; multibacillary infections are common. We reported pediatric cases with WHO-multibacillary lepromatous type morbus Hansen. We discovered several hypopigmented patches on the right upper arm, anesthesia-filled claw hands, and infiltrates in the ears in one 17- year-old child. In the other case (13-year-old), we discovered reduced sensory sensitivity in nearly every area of the body, infiltrates on the ear, and numerous irregular hypopigmented plaques on the belly, lumbar, and gluteus. Both patients had positive acidfast bacteria. After three months of treatment, multibacillary leprosy patients began to show improvement. Low socioeconomic and environmental conditions, as well as a history of intimate contact with leprosy patients, have been identified as risk factors.

AN UPDATE TREATMENT OF ERITEMA NODOSUM LEPROSUM

Leprosy, also called Morbus Hansen, is a contagious skin disease caused by the bacterium Mycobacterium Leprae. It takes 2–5 years for the disease to show up, and it takes 2–3 weeks for the cells to divide. Leprosy reaction is a sudden event that happens during the long-term course of the disease. It causes skin sores to become very red and swollen. This disease can show up before, during, or after treatment for leprosy, and it can happen to 30–50% of people with leprosy. Reactions to leprosy happen when both cellular and humoral immune responses go wrong. Type 2 or Eritema Nodosum Leprosum (ENL) reactions have a clinical picture of the appearance of nodules in the skin. This reaction occurs in leprosy patients with very large numbers of bacteria, namely the multibacillary type. In contrast to the reversal reaction, the humoral immune response plays an important role in the pathogenesis of ENL. Large amounts of bacterial antigen concentrations in tissues will increase IgM and IgG titers in multibacillary leprosy patients. Th2 cell activation will stimulate the production of IL-4 and IL-10, thereby increasing the humoral immune response and increasing the production of B lymphocytes. The goals of the treatment are to control the inflammation, ease the pain, and stop new cases from happening. In weak cases of ENL, the best way to treat it is to rest and take anti-inflammatory drugs. Most of the time, people take aspirin, which is an anti-inflammatory drug. Other drugs have also been tried, including nonsteroidal anti-inflammatory drugs (NSAIDs), colchicine, oral zinc, pentoxifylline, and chloroquine. But each of the current treatment options has its own downsides and limitations, and the best one must be carefully chosen for each case.

Positive Correlation of Plasma Interleukin-6 Levels with Immunoglobulin M (IgM) Anti Phenolic Glycolipid-1 (PGL-1) Levels in Household Contacts of Multibacillary Leprosy Patients

Background/aim: Leprosy is a chronic infectious disease that can be cured but causes high morbidity because it causes disability. Leprosy is caused by the bacterium Mycobacterium leprae, an obligate intracellular virulence factor in the form of phenolic glycolipid-1 (PGL-1), a capsule component in M. leprae. This study aims for known Increased IL-6 correlation with IgM anti PGL-1 level in household contacts of multibacillary leprosy patient. Methods: Analytical observational study with a cross-sectional approach. The sample consisted of 42 household contact persons and 22 non-contact samples aged 18-65 years who were selected through consecutive sampling according to the inclusion and exclusion criteria. Samples of 3 millilitres of venous blood were taken, which would then be examined for PGL-1 and IL-6 with the ELISA kit. Data analysis with SPSS version 23 and p-value &lt;0.05 was significant Results: The average anti-PGL-1 IgM level in the household contact group was 636.76±288.45 u/ml, while in the non-contact group, it was 211.00±237.43 u/ml. The average IL-6 level in the household contact group was 20.28 ± 12.54 pg/ml, while in the non-contact group, it was 8.29 ± 2.88 pg/ml. There was a significant difference in the levels of IL-6 and anti-PGL-1 IgM in the household contact and non-contact groups (p&lt;0.001). This study found a strong positive relationship (r=0.620, p&lt;0.001) between IL-6 levels and anti-PGL-1 IgM levels. Conclusion: IgM anti-PGL-1 and IL-6 levels in household contacts were higher than in non-contacts, with a significant difference. There is a significant positive relationship between IL-6 and anti-PGL-1 IgM levels. Keywords: leprosy, contacts, IgM anti-PGL-1, IL-6

High levels of zinc (Zn) as a protective factor and negatively correlated with IgM anti PGL-1 levels among household contact with multibacillary leprosy patients

Background: Close contacts may develop subclinical leprosy with no symptoms but with M. leprae in their blood. Zinc is said to help in the prevention of M. leprosy infection. This study aimed to determine the correlation between zinc (Zn) serum levels and IgM anti PGL-1 levels in household contacts of multibacillary type leprosy patients.&#x0D; Methods: This study is cross-sectional and involves 48 leprosy subjects (33 multibacillary leprosy household contact and 15 non-contact subjects). Subjects were selected based on the inclusion and exclusion criteria. Zinc and IgM anti PGL-1 levels were derived from venous blood examined with ELISA.&#x0D; Results: This study showed the mean serum zinc level of the contact groups of 60.88 ± 15.92 μg/dl and the non-contact group of 90.00 ± 7.61 μg / dl (p &lt;0.001; CI 95%: 20.38 - 37.85). The median (interquartile range) serum levels of IgM anti PGL-1 in the contact groups was 613.00 (40 - 1433) u/ml and the non-contact groups was 99.00 (14 - 695) u/ml (p = 0.001; 95% CI: 154.162 – 610.116). The correlation analysis between serum zinc levels and IgM anti PGL-1 serum levels r= 0.644 (p&lt;0.05). A high serum zinc level is a protective factor against high IgM anti PGL-1 levels in household contact with leprosy (PR: 0.47; 95% CI: 0.20-1.09).&#x0D; Conclusion: Zinc levels negatively correlate with IgM anti PGL-1 levels in subjects with household contact with multibacillary type leprosy. A high serum zinc level protects against high IgM anti PGL-1 levels.

Existence of viable Mycobacterium leprae in natural environment and its genetic profiling in a leprosy endemic region

IntroductionMolecular epidemiology of leprosy is very important to study leprosy transmission dynamics and to enhance our understanding of leprosy in endemic areas by utilizing the molecular typing method. Nowadays our understanding of leprosy transmission dynamics has been refined by SNP typing and VNTR marker analysis of M. leprae strains.ObjectiveThis study was carried out to find out the presence of viable M. leprae in the soil and water samples from residing areas of leprosy patients staying in different blocks of Purulia district of West Bengal, understanding their genotypes and compared with that of M. leprae present in patients.Material and methodsSlit-skin smear (SSS) samples (n=112) were collected from the active multibacillary leprosy patients from different blocks of leprosy endemic area. Soil samples (n=1060) and water samples (n=620) were collected from residing areas of leprosy patients. SNP subtyping was performed by PCR followed by sequencing. Multiplex PCR was performed using fifteen ML-VNTR loci and results were analysed.ResultsWe observed high PCR positivity in soil samples (344 out of 1060; 32%) and water samples (140 out of 620; 23%). These PCR positive samples when further screened for viability, it was observed that 150 soil samples (44%) and 56 water samples (40%) showed presence of 16S rRNA. SNP typing of M. leprae revealed presence of predominantly type 1. SNP subtype 1D (83%) was most prevalent in all the blocks of Purulia followed by subtype 1C (15%) and subtype 1A (2%). SNP subtype 2F was noted in only one sample. SNP and VNTR combination showed presence of similar strain type in certain pockets of Purulia region which was responsible for transmission.ConclusionPresence of viable M. leprae in the environment, and presence of SNP Type 1 M. leprae in patients and environment suggests both environment and patients play a role in disease transmission.

Evaluation of altered patterns of tactile sensation in the diagnosis and monitoring of leprosy using the Semmes-Weinstein monofilaments.

Leprosy neuropathy is the most common peripheral neuropathy of infectious etiology worldwide; it is characterized as asymmetric and focal multiple mononeuropathy. Semmes-Weinstein monofilament (SWM) test is a simple method to assess sensory nerve function. In this prospective cohort study, a dermatologist carried out hands and feet tactile sensation test with SWM in 107 multibacillary leprosy patients at diagnosis and in 76 patients at the end of treatment from 2016 to 2019. At diagnosis, 81/107 (75.7%) patients had some degree of functional disability, and 46 (43%) of them had altered SWM-test in the hands and 94 (87.9%) of them in the feet. After one year of multibacillary multidrug therapy, the disability decreasing to 44/76 patients (57.9%) and decreasing of the percentual of patients with altered SWM-test to 18% for the hands, and to 28.7% for the feet. At the end of treatment, the number of SMW-test points presented improvement in the hands of 22 (28.9%) patients, and in the feet of 47 (61.8%). In the hands, by SWM-test, only the radial nerve point demonstrated a significant asymmetry, while in the feet, the difference between the sum of altered SWM-test points showed significant asymmetry between both sides, highlighting the tibial nerve for the establishment of asymmetric leprosy neuropathy. In Spearman's correlation analysis, a positive correlation with statistical significance was observed between the number of hands and feet SWM altered points at diagnosis and the degree of disability at diagnosis (0.69) and at the end of the treatment (0.80). The patterns of hands and feet tactile sensation at diagnosis and their consequent modifications with the anti-leprosy drugs define the bacterial etiology of neuropathy, an important tool for the clinical diagnosis and follow up of the disease, highlighting the tibial nerve findings, the most affected nerve among leprosy patients by SWM-test, with significant asymmetry and focality impairments.

Downmodulation of Regulatory T Cells Producing TGF-β Participates in Pathogenesis of Leprosy Reactions.

Leprosy reactions are an acute and systemic manifestation, which occurs suddenly, can be severe and lead leprosy patients to disability. Reactional episodes are observed among half of the multibacillary patients, mainly in borderline lepromatous and lepromatous forms. They may begin at any time during multidrug therapy, and even before the treatment. Physical disabilities, which are the source of extreme suffering and pain for patients, occur in progression of the cellular immune response associated with a reaction and are still poorly understood. Thus, this work aimed to phenotypically and functionally characterize CD4+ and CD8+ Treg cells ex vivo and in response to Mycobacterium leprae (ML). We studied 52 individuals, including 18 newly diagnosed and untreated multibacillary leprosy patients, 19 reactional multibacillary patients (Type I or Type II episodes) and 15 healthy volunteers, included as controls, all residents of the city of Rio de Janeiro. The functional activity and frequencies of these cells were evaluated through multiparametric flow cytometry. In addition, the production of cytokines in supernatant from peripheral blood mononuclear cell cultures was also investigated against ML by enzyme-linked immunosorbent assay. Our results showed a decrease in CD4+TGF-β+ Treg and CD8+ TGF-β+ Treg in leprosy multibacillary patients during both types of reactional episodes. Alterations in the cytokine profile was also observed in Type II reactions, along with upregulation of IL-17 and IL-6 in supernatant. Thus, our study suggests that downregulation of Treg cells is related with both classes of reactional episodes, improving our understanding of immune hyporesponsiveness in multibacillary patients and hyperesponsiveness in both reactions.

The Type I Interferon Pathway Is Upregulated in the Cutaneous Lesions and Blood of Multibacillary Leprosy Patients With Erythema Nodosum Leprosum.

In leprosy patients, acute inflammatory episodes, known as erythema nodosum leprosum (ENL), are responsible for high morbidity and tissue damage that occur during the course of Mycobacterium leprae infection. In a previous study, we showed evidence implicating DNA-sensing via TLR9 as an important inflammatory pathway in ENL. A likely important consequence of TLR9 pathway activation is the production of type I interferons (IFN-I) by plasmacytoid dendritic cells (pDCs), also implicated in the pathogenesis of several chronic inflammatory diseases. In this study, we investigated whether the IFN-I pathway is activated during ENL. Blood samples and skin lesions from multibacillary patients diagnosed with ENL were collected and the expression of genes of the IFN-I pathway and interferon-stimulated genes were compared with samples collected from non-reactional multibacillary (NR) patients. Whole blood RNAseq analysis suggested higher activation of the IFN-I pathway in ENL patients, confirmed by RT-qPCR. Likewise, significantly higher mRNA levels of IFN-I-related genes were detected in ENL skin biopsies when compared to NR patient lesions. During thalidomide administration, the drug of choice for ENL treatment, a decrease in the mRNA and protein levels of some of these genes both in the skin and blood was observed. Indeed, in vitro assays showed that thalidomide was able to block the secretion of IFN-I by peripheral blood mononuclear cells in response to M. leprae sonicate or CpG-A, a TLR9 ligand. Finally, the decreased frequencies of peripheral pDCs in ENL patients, along with the higher TLR9 expression in ENL pDCs and the enrichment of CD123+ cells in ENL skin lesions, suggest the involvement of these cells as IFN-I producers in this type of reaction. Taken together, our data point to the involvement of the pDC/type I IFN pathway in the pathogenesis of ENL, opening new avenues in identifying biomarkers for early diagnosis and new therapeutic targets for the better management of this reactional episode.

Arginase 1 is a marker of protection against illness in contacts of leprosy patients

Leprosy household contacts are generally more prone to develop the disease compared to the general population. Previous studies have demonstrated that genes related to the alternative activation (M2) profile in macrophages are associated with the increased bacillary load in multibacillary leprosy patients (MB), and that contacts of MB patients have a higher risk of contracting the disease. In addition, positive serological responses to PGL-1 or LID-1 are associated with a higher risk of disease. We performed a 5-year follow-up of contacts of leprosy patients and evaluated the pattern of gene and protein expression in cells from contacts that developed leprosy during this period. Leprosy household contacts had decreased soluble CD163 and heme oxygenase 1 (HO-1) serum levels when compared with healthy donors and leprosy patients. In contrast, arginase 1 activities were higher in contacts when compared with both healthy donors and leprosy patients. Of the contacts, 33 developed leprosy during the follow-up. Gene expression analysis revealed reduced ARG1 expression in these contacts when compared with contacts that did not develop disease. Arginase activity was a good predictive marker of protection in contacts (sensitivity: 90.0%, specificity: 96.77%) and the association with serology for anti-PGL-1 and anti-LID-1 increased the sensitivity to 100%. Altogether, the data presented here demonstrate a positive role of arginase against leprosy and suggest that the evaluation of arginase activity should be incorporated into leprosy control programs in order to aid in the decision of which contacts should receive chemoprophylaxis.

Correlation between Interleukin 10 Serum Levels and Severity of Peripheral Neuropathy in Multibacillary Leprosy Patients a Research Article

ABSTRACT Background of the ResearchLeprosy is a chronic infectious peripheral neuropathy caused by Mycobacterium leprae. Clinical presentations of leprosy neuropathy are obtained when the nerve damage is more than 30% of peripheral nerve fibers. The anti-inflammatory property of interleukin 10 is supposed to protect tissue from the damage caused by proinflammatory cytokines. Instead, it causes extensive nerve damage due to suppression of the cellular immune system, failure of forming granuloma to limit the growth of Mycobacterium leprae. As a result, it multiplies and spreads. Purpose of the ResearchThe purpose of the research is to analyze the correlation between interleukin levels 10 serum and the severity of peripheral neuropathy. Methods of the ResearchThe method used is cross-sectional research on 30 subjects with multibacillary leprosy aged 18-65 at the Neurology Department of Kelet Hospital Jepara from September to October 2021. The questionnaire was used to collect data of the patients. Interleukin level 10 serum evaluation is assessed by examining blood serum. Data analysis used Spearman Correlation Test. The significant result occurs if it is p&lt;0.05. Research resultPatients with multibacillary leprosy are mostly men (73.3%). There is also a strong significant correlation between interleukin 10 levels and the severity of peripheral neuropathy (p&lt;0.001) with rho (0.7) ConclusionThere is a strong and significant correlation between serum interleukin 10 levels and the severity of peripheral neuropathy in multibacillary leprosy Keywords: multibacillary leprosy, interleukin 10, peripheral neuropathy

Standardization of SYBR Green-Based Real-Time PCR Through the Evaluation of Different Thresholds for Different Skin Layers: An Accuracy Study and Track of the Transmission Potential of Multibacillary and Paucibacillary Leprosy Patients.

The development of new molecular techniques is essential for the early diagnosis of leprosy. Studies in the field have failed to elucidate the performance of these tests in clinical practice. We aimed to design a new primer pair for the repetitive element (RLEP) target of Mycobacterium leprae and to test the accuracy of SYBR green-based real-time PCR through the evaluation of different thresholds for different skin layers. We also aimed to track the transmission potential of multibacillary and paucibacillary leprosy patients. The in vitro validation of our reaction resulted in a quantification limit of 0.03 bacilli. We then conducted a cross-sectional/cohort-based study of diagnostic accuracy. Patients were included, and skin samples were divided into four layers: epidermis, superior dermis, inferior dermis, and hypodermis. We also quantified M. leprae in nasal swabs of the included patients and compared the results to the number of household contacts also diagnosed with leprosy. One hundred patients with a clinical presentation compatible with leprosy were allocated to the leprosy or control group. Although the parasite load was greater in the superior and inferior dermis, M. leprae DNA was found in all skin layers. The best sensitivity was observed for the superior dermis using the presence of any quantifiable bacillus DNA as the threshold [sensitivity=59.26% (95% CI=45.97–71.32)]. In the epidermis, setting 1 quantifiable bacillus as the threshold resulted in 100% specificity (95% CI=92.29–100). The number of bacilli found in nasal swabs was not significantly related to the number of household contacts also diagnosed with leprosy. Paucibacillary patients tested positive only for bacillus fragments in nasal swabs but not for the entire bacilli. We can conclude that superficial biopsies might result in sensitivity loss, although different skin sample types will have little influence on the final accuracy. In contrast, threshold changes greatly influence these properties. Paucibacillary patients may not be a relevant source of disease transmission.

The Demography, Clinical Characteristics, and White Blood Analysis of Leprosy Reactions in Multibacillary Leprosy: A Retrospective Study

Background: Leprosy is a neglected tropical disease caused by chronic granulomatous infection of Mycobacterium leprae. Indonesia ranks third in new case findings, with 84% of the case being multibacillary (MB) leprosy. MB leprosy cases have a higher risk of leprosy reactions and physical disabilities that decrease quality of life. Purpose: To determine the demographic, clinical characteristics, and white blood analysis of newly diagnosed MB leprosy patients, especially concerning leprosy reactions. Methods: This is a descriptive retrospective study with a cross-sectional design that describe the following data: domicile, gender, age, treatment status, disabilities, body mass index (BMI); bacterial index (BI), morphological index (MI), white blood cell (WBC) and differential counts, and thrombocyte count. Result: This study included 176 adult MB cases, predominantly male aged 20–39 years old with average BMI, lived in Surabaya with negative history of multi-drug therapy, disability, BI, nor MI. The grade 2 disability (G2D) percentage in this study setting than in Indonesia (10.7% vs. 6.43%). The WBCs, especially neutrophil count, was higher in T2R group. Monocyte and lymphocyte counts were relatively similar. There was an increase in thrombocyte count in leprosy reaction groups. Conclusion: MB leprosy in the endemic area, which is more commonly found in productive-aged male, displayed higher G2D than global Indonesia population. Thus denotes the importance of active case findings. The difference in blood analysis characteristics between MB leprosy with and without reactions may serve as the foundation for future study.

RLEP LAMP for the laboratory confirmation of leprosy: towards a point-of-care test

BackgroundNucleic acid-based amplification tests (NAAT), above all (q)PCR, have been applied for the detection of Mycobacterium leprae in leprosy cases and household contacts with subclinical infection. However, their application in the field poses a range of technical challenges. Loop-mediated isothermal amplification (LAMP), as a promising point-of-care NAAT does not require sophisticated laboratory equipment, is easy to perform, and is applicable for decentralized diagnosis at the primary health care level. Among a range of gene targets, the M. leprae specific repetitive element RLEP is regarded as highly sensitive and specific for diagnostic applications. MethodsOur group developed and validated a dry-reagent-based (DRB) RLEP LAMP, provided product specifications for customization of a ready-to-use kit (intended for commercial production) and compared it against the in-house prototype. The assays were optimized for application on a Genie® III portable fluorometer. For technical validation, 40 “must not detect RLEP” samples derived from RLEP qPCR negative exposed and non-exposed individuals, as well as from patients with other conditions and a set of closely related mycobacterial cultures, were tested together with 25 “must detect RLEP” samples derived from qPCR confirmed leprosy patients. For clinical validation, 150 RLEP qPCR tested samples were analyzed, consisting of the following categories: high-positive samples of multibacillary (MB) leprosy patients (> 10.000 bacilli/extract), medium-positive samples of MB leprosy patients (1.001–10.000 bacilli/extract), low-positive samples of MB leprosy patients (1–1.000 bacilli/extract), endemic controls and healthy non-exposed controls; each n = 30. ResultsTechnical validation: both LAMP formats had a limit of detection of 1.000 RLEP copies, i.e. 43–27 bacilli, a sensitivity of 92% (in-house protocol)/100% (ready-to-use protocol) and a specificity of 100%. Reagents were stable for at least 1 year at 22 °C. Clinical validation: Both formats showed a negativity rate of 100% and a positivity rate of 100% for high-positive samples and 93–100% for medium positive samples, together with a positive predictive value of 100% and semi-quantitative results. The positivity rate for low-positive samples was 77% (in-house protocol)/43% (ready-to-use protocol) and differed significantly between both formats. ConclusionsThe ready-to-use RLEP DRB LAMP assay constitutes an ASSURED test ready for field-based evaluation trials aiming for routine diagnosis of leprosy at the primary health care level.

Hubungan Asupan Vitamin D Dan Selenium Dengan Indeks Bakteriologi Penderita Kusta Tipe MB (Multibacillary) : Systematic Literature Review

Background : As one of the infectious diseases that have a high prevalence until now, leprosy is a chronic transmission disease caused by infection with Mycobacterium Leprae. Apart from the standard MDT (Multidrug Therapy), nutritional support is a crucial component to restrict further infection severity. Vitamin D and selenium play essential roles as the primary defence against susceptibility. Methods : The systematic literature review was conducted using three international databases : PubMed, ResearchGate, and DOAJ, which were published between 2010 – 2020 using English or Indonesian. A total of 8 articles (2 articles identifying the correlation of selenium with leprosy, and 6 articles identifying the correlation of vitamin D with leprosy) using cross- sectional analytic and case-control analytic studies were included in this review. Results : The average serum levels of vitamin D and selenium in leprosy patients were lower than in healthy controls. This case was also found in Multibacillary leprosy patients compared to Paucibacillary leprosy patients. This serum levels can affect the bacteriological index, which is the higher the serum levels, the lower the bacteriological index. Disscusion : Serum vitamin D and selenium levels are affected by vitamin D and selenium intake, which can lead to a reduction in bacteriological index across all multibacillary leprosy patients. There is a negative correlation between vitamin D, selenium, and bacteriological index in patients with MB (Multibacillary) leprosy.Keywords:Leprosy, Vitamin D, Selenium, Bacteriological Index

Effect of Relationship between Protein Oxidation and Total Thiols in Progression of Leprosy

Objective: In several countries including India, leprosy is an older disease &amp; until now continuous to be an important health issue. Leprosy is a chronic granulomatous disease, the causative agent for which is mycobacterium leprae. Due to imbalance between natural antioxidants and oxidative stress is significant event occurs that influences the pathogenesis of leprosy. Considering this the study was carried out to find, what type of relationship between MDA, Protein carbonyl and thiols as disease advances.&#x0D; Methods: 50 diagnosed leprosy patients &amp; 50 healthy controls were included in this study. In leprosy group, 16 were Paucibacillary (PB) and 34 were Multibacillary (MB) type leprosy patients.&#x0D; Results: Serum Protein carbonyl and serum Malondialdehyde increased in leprosy patients than controlling. Further analysis reveals that the serum malondialdehyde was significantly increased in MB leprosy patients than PB leprosy patients. On the other hand, our proposed system significantly lowers the total thiols in leprosy patients when compared with controls. Serum total thiols were lower in MB leprosy than PB leprosy patients. Among leprosy patients the negative correlation of Malondialdehyde with total thiol and Protein carbonyl with total thiols was observed.&#x0D; Conclusion: Thus, as protein oxidation and lipid per oxidation in leprosy increases, the total thiols decreases which may lead to modifications of protein and lipids that are responsible for severity of disease hence therapy aimed at reducing generation of free radicals that can result in oxidative stress and as a result a modification of proteins and lipids.