Spatial distribution pattern of new leprosy cases under 15 years of age and their contacts in Sobral, Ceará, Brazil.

This study's objective was to analyze the spatial distribution pattern of new leprosy cases under 15 years and their contacts. A cross-sectional descriptive study covering sociodemographic characteristics and spatial analysis was carried out. The participants were from the city of Sobral, Ceará and the study was conducted between August 2014 and September 2015. Contacts were identified by the persons responsible for the children. Seropositivity was determined with the NDO-LID antigen, and positive cases were plotted on Voronoi polygons. Nine new cases of leprosy under 15 years of age have been found. The average number of people living with the cases was higher than the number of people living with non-household contacts. All household contacts were aware of other leprosy cases and had a higher rate of seropositive tests than non-household contacts. The index cases lived in the poorest regions of the municipality and hyper-endemic areas. Spatial analysis revealed a cluster of subclinical infection within a radius of 102 meters, suggesting that non-household transmission is related to proximity with seropositive individuals. In conclusion, the search for new leprosy cases cannot be restricted to household contacts.

Spatial distribution pattern of new leprosy cases under 15 years of age and their contacts in Sobral, Ceará, Brazil / Distribuição espacial de novos casos de hanseníase em menores de 15 anos e de seus contatos em Sobral, Ceará, Brasil

Abstract This study's objective was to analyze the spatial distribution pattern of new leprosy cases under 15 years and their contacts. A cross-sectional descriptive study covering sociodemographic characteristics and spatial analysis was carried out. The participants were from the city of Sobral, Ceará and the study was conducted between August 2014 and September 2015. Contacts were identified by the persons responsible for the children. Seropositivity was determined with the NDO-LID antigen, and positive cases were plotted on Voronoi polygons. Nine new cases of leprosy under 15 years of age have been found. The average number of people living with the cases was higher than the number of people living with non-household contacts. All household contacts were aware of other leprosy cases and had a higher rate of seropositive tests than non-household contacts. The index cases lived in the poorest regions of the municipality and hyper-endemic areas. Spatial analysis revealed a cluster of subclinical infection within a radius of 102 meters, suggesting that non-household transmission is related to proximity with seropositive individuals. In conclusion, the search for new leprosy cases cannot be restricted to household contacts.

Resumo O objetivo deste estudo foi analisar o padrão de distribuição espacial de novos casos de hanseníase em menores de 15 anos e seus contatos. Estudo transversal, descritivo, abrangendo características sociodemográficas e análise espacial. Os participantes eram de Sobral, Ceará e o estudo foi realizado entre agosto de 2014 e setembro de 2015. Os contatos foram identificados pelos responsáveis pelas crianças. A soropositividade foi determinada com o antígeno NDO-LID e os casos positivos foram plotados em polígonos de Voronoi. Nove novos casos de hanseníase em menores de 15 anos foram encontrados. O número médio de pessoas que conviviam com os casos foi superior ao número de pessoas que conviviam com contatos não domiciliares. Todos os contatos domiciliares sabiam de outros casos de hanseníase e apresentaram maior taxa de testes soropositivos do que os contatos não domiciliares. Os casos índice residiam nas regiões mais pobres do município e em áreas hiperendêmicas. A análise espacial revelou um agrupamento de infecção subclínica em um raio de 102 metros, sugerindo que a transmissão não domiciliar está relacionada à proximidade com indivíduos soropositivos. Concluindo, a busca por novos casos de hanseníase não pode se restringir aos contatos domiciliares.

Genotyping of Mycobacterium leprae for better understanding of leprosy transmission in Fortaleza, Northeastern Brazil.

Leprosy is endemic in large part of Brazil with 28,761 new patients in 2015, the second largest number worldwide and reaches 9/10.000 in highly endemic regions and 2.7/10.000 in the city of Fortaleza, Ceará, Northeast Brazil. For better understanding of risk factors for leprosy transmission, we conducted an epidemiologic study supplemented by 17 locus VNTR and SNP 1-4 typing of Mycobacterium leprae in skin biopsy samples from new multibacillary (MB) patients diagnosed at a reference center in 2009 and 2010. Among the 1,519 new patients detected during the study period, 998 (65.7%) were MB and we performed DNA extraction and genotyping on 160 skin biopsy samples, resulting in 159 (16%) good multilocus VNTR types. Thirty-eight of these patients also provided VNTR types from M. leprae in nasal swabs. The SNP-Type was obtained for 157 patients and 87% were of type 4. Upon consideration all VNTR markers, 156 different genotypes and three pairs with identical genotypes were observed; no epidemiologic relation could be observed between individuals in these pairs. Considerable variability in differentiating index (DI) was observed between the different markers and the four with highest DI [(AT)15, (TA)18, (AT)17 and (GAA)21] frequently demonstrated differences in copy number when comparing genotypes from both type of samples. Excluding these markers from analysis resulted in 83 genotypes, 20 of which included 96 of the patients (60.3%). These clusters were composed of two (n = 8), three (n = 6), four (n = 1), five (n = 2), six (n = 1), 19 (n = 1) and 23 (n = 23) individuals and suggests that recent transmission is contributing to the maintenance of leprosy in Fortaleza. When comparing epidemiological and clinical variables among patients within clustered or with unique M. leprae genotypes, a positive bacterial index in skin biopsies and knowledge of working with someone with the disease were significantly associated with clustering. A tendency to belong to a cluster was observed with later notification of disease (mean value of 3.4 months) and having disability grade 2. A tendency for lack of clustering was observed for patients who reported to have lived with another leprosy case but this might be due to lack of inclusion of household contacts in the study. Although clusters were spread over the city, kernel analysis revealed that some of the patients belonging to the two major clusters were spatially related to some neighborhoods that report poverty and high disease incidence in children. Finally, inclusion of genotypes from nasal swabs might be warranted. A major limitation of the study is that sample size of 160 patients from a two year period represents only 15% of the new patients and this could have weakened statistical outcomes. This is the first molecular epidemiology study of leprosy in Brazil and although the high clustering level suggests that recent transmission is the major cause of disease in Fortaleza; the existence of two large clusters needs further investigation.

Intrapatient comparison of Mycobacterium leprae by VNTR analysis in nasal secretions and skin biopsy in a Brazilian leprosy endemic region.

Background: This study compares the strains of genotypes of M. leprae from nasal secretions (NS) and skin biopsy (SB) in the same patient, supplementing conventional epidemiology to gain insight into the infection of leprosy in Fortaleza, Brazil. Methods: The sample consisted of 38 newly diagnosed leprosy patients attending the National Reference Center of Dermatology Dona Libania (CDERM), in Fortaleza, who tested positive for M. leprae by PCR in DNA extracts of nasal secretions. DNA was also extracted from skin biopsy (SB) scrapings of each patient and used for multiplex PCR amplification of M. leprae VNTR loci. The number of repeats at 15 loci were determined by the fragment length analysis method. Results: Locus VNTR genotypes were achieved in 38 NS, and in 38 SB specimens. M. leprae strains differed in their genotypes in paired specimens in all but two of 38 patients. The genotype similarity in the remainder ranged from 53% to 87%. Conclusion: M. leprae 15 VNTR loci genotypes of paired nasal and biopsy skin samples from five patients were identical, while as many as seven loci differed in the 33 other patients. When the NS and biopsy genotypes were pooled and compared, it was found that there was a great variability among different VNTR markers. It is important to investigate other molecular markers suitable for typing genetic variations of the bacilli.

Seropositivity to anti-phenolic glycolipid-I in leprosy cases, contacts and no known contacts of leprosy in an endemic and a non-endemic area in northeast Brazil.

The seroprevalence rates of IgM anti-phenolic glycolipid-I (PGL-I) antibodies in four study groups with differing exposure to Mycobacterium leprae in Ceará, Brazil were investigated between March 2005 and August 2006. The first three groups in a high prevalence area included 144 cases of leprosy, their 380 contacts and 317 participants with no known leprosy contact. The fourth group in a low prevalence area consisted of 87 participants with no known leprosy contact living in an area in which no cases of leprosy had been reported in the previous 6 months. Seropositivity and levels of IgM antibodies to PGL-I were investigated using ELISA. The seropositivity levels of anti-PGL-I among the different clinical forms of leprosy cases were 61% for lepromatous, 25% for tuberculoid and 27% indeterminate. The levels of anti-PGL-I antibodies in the endemic area differentiated leprosy cases from non-cases. However, the seropositivity was similar among contact cases (15.8%) and no known leprosy contact cases from high (15.1%) and low (13.8%) prevalence areas. The seropositivity of both contacts and no known contacts was much higher than previously reported among no known contacts in other endemic areas. The study indicates that anti-PGL-I antibodies are not useful as immunological markers of household leprosy contacts and no known leprosy contacts in endemic areas.

Mycobacterium tuberculosis Rv1419 encodes a secreted 13 kDa lectin with immunological reactivity during human tuberculosis.

In this study, we have identified a secreted 13 kDa lectin from Mtb (Mtb, Mycobacterium tuberculosis; sMTL-13) by homology search of a non-redundant lectin database. Bioinformatic analysis revealed that sMTL-13 belongs to the ricin-type beta-trefoil family of proteins containing a Sec-type signal peptide present in Mtb complex species, but not in non-tuberculous mycobacteria. Following heterologous expression of sMTL-13 and generation of an mAb (clone 276.B7/IgG1kappa), we confirmed that this lectin is present in culture filtrate proteins from Mtb H37Rv, but not in non-tuberculous mycobacteria-derived culture filtrate proteins. In addition, sMTL-13 leads to an increased IFN-gamma production by PBMC from active tuberculosis (ATB) patients. Furthermore, sera from ATB patients displayed high titers of IgG Ab against sMTL-13, a response found to be decreased following successful anti-tuberculosis therapy. Together, our findings reveal a secreted 13 kDa ricin-like lectin from Mtb, which is immunologically recognized during ATB and could serve as a biomarker of disease treatment.

Viral Hepatitis C in a leading Brazilian hospital: epidemiological factors and genotyping

The hepatitis C virus (HCV) is classified into six different genotypes and their distribution is different throughout the world. Epidemiologic studies are important to determine several characteristics of the virus, as well as the disease. This study analysed the prevalence of HCV and its genotypes among patients from a leading hospital in Ceará, which is located in Northeast Brazil. A total of 119 anti-HCV-seropositive patients, each having previously completed a questionnaire about risk behaviours related to HCV infection were tested for HCV infection using a qualitative HCV polymerase chain reaction (PCR) assay and genotyping by restriction fragment length polymorphism (RFLP). The detection was based on amplifying of the non-coding 5' region. Of the 119 patients, 95 showed positive results in the qualitative HCV test. History of surgery was the most reported risk factor, followed by the use of drugs, having tattoos, undergoing haemodialysis and occupational exposure. Genotype 1 was the most prevalent (46.9 percent), followed by genotype 3 (34.4 percent) and 2 (8.3 percent). The genotype distribution was similar for all of the various risk behaviours.

O vírus da hepatite C (VHC) é classificado em seis genótipos diferentes e sua distribuição é diferente em todo o mundo. Os estudos epidemiológicos são importantes para determinar várias características sobre o vírus, bem como da doença. Este estudo analisou a prevalência do VHC e seus genótipos em pacientes atendidos em hospital de referência no Ceará, o qual é localizado no nordeste do Brasil. Um total de 119 pacientes, os quais eram soropositivos anti-VHC, preencheram questionários sobre fatores de risco relacionados à infecção pelo VHC e foram testados quanto à infecção ao VHC usando o teste da reação da polimerase em cadeia (PCR) qualitativo para VHC e genotipagem por "restriction fragment length polymorphism" (RFLP). A detecção foi baseada na amplificação da região não codificante 5'. Dos 119 pacientes, 95 mostraram resultados positivos no teste qualitativo para VHC. A história prévia de cirurgia foi o fator de risco mais relatado, seguido pelo uso de drogas, ter tatuagem, ter sido submetido à hemodiálise e risco ocupacional. O genótipo 1 foi o mais prevalente (46,9 por cento), seguido pelo genótipo 3 (34,4 por cento) e 2 (8,3 por cento). A distribuição dos genótipos foi similar entre os vários fatores de risco analisados.

The AraC family transcriptional regulator Rv1931c plays a role in the virulence of Mycobacterium tuberculosis.

A Mycobacterium tuberculosis strain disrupted in the AraC homologue Rv1931c was isolated. The mutant strain exhibited reduced survival both in macrophages and in a mouse infection model, with survival being restored on complementation with the Rv1931c gene. These results suggest that Rv1931c regulates genes important for virulence of M. tuberculosis.

Genome structure in the vole bacillus, Mycobacterium microti, a member of the Mycobacterium tuberculosis complex with a low virulence for humans.

Mycobacterium microti, a member of the Mycobacterium tuberculosis complex, is phylogenetically closely related to M. tuberculosis, differing in a few biochemical properties. However, these species have different levels of virulence in different hosts; most notably M. microti shows lower virulence for humans than M. tuberculosis. This report presents genomic comparisons using DNA microarray analysis for an extensive study of the diversity of M. microti strains. Compared to M. tuberculosis H37Rv, 13 deletions were identified in 12 strains of M. microti, including the regions RD1 to RD10, which are also missing in Mycobacterium bovis BCG. In addition, four new deleted regions, named MiD1, RD1beta, MiD2 and MiD3, were identified. DNA sequencing was used to define the extent of most of the deletions in one strain. Although RD1 of M. bovis BCG and M. microti is thought to be crucial for attenuation, in this study, three of the four M. microti strains that were isolated from immunocompetent patients had the RD1 deletion. In fact, only the RD3 deletion was present in all of the strains examined, although deletions RD7, RD8 and MiD1 were found in almost all the M. microti strains. These deletions might therefore have some relation to the different host range of M. microti. It was also noticeable that of the 12 strains studied, only three were identical; these strains were all isolated from immunocompetent humans, suggesting that they could have arisen from a single source. Thus, this study shows that it is difficult to ascribe virulence to any particular pattern of deletion in M. microti.