RESUMO
We aimed to identify an unique host transcriptional signature in peripheral blood mononuclear cells (PBMCs) in response to Mycobacterium leprae antigens to distinguish between patients with leprosy and non-leprosy controls for early diagnosis of the disease. Sixteen individuals were enrolled in the discovery cohort [eight patients with leprosy, comprising four multibacillary (MB) and four paucibacillary (PB); and eight non-leprosy controls, comprising four healthy house contacts (HHCs) and four endemic controls (ECs)]. The differences in the transcriptome response of PBMCs to M. leprae sonicate antigen were evaluated between leprosy patients and non-leprosy controls, and 12 differentially expressed genes (CCL2/MCP-1, IL-8, JAKM, ATP, ND1, SERP, FLJ10489, LINC00659, LOC34487, LOC101928143, MIR22, and NCF1C) were identified. The accuracy of the 12 differentially expressed genes was further validated for the diagnosis of leprosy using real-time quantitative PCR in 82 individuals (13 MB, 10 PB, 37 HHCs, and 22 ECs) in the validation cohort. We found that a 5 gene signature set IL-8, CCL2/MCP-1, SERP, LINC00659 and FLJ10489 had a suitable performance in discriminating leprosy from ECs. In addition, elevated expression of IL-8, CCL2/MCP-1, SERP and LINC00659 was associated with MB diagnosis compared with ECs, whereas increased expression of IL-8, CCL2/MCP-1, SERP and FLJ10489 was found to be useful biomarkers for PB diagnosis from ECs. Moreover, we found decreased expression of NCF1C among leprosy patients could distinguish leprosy from HHCs, whereas higher expression of CCL2 among MB than PB could distinguish different leprosy patients. In conclusion, among the 12 candidate host genes identified, a three gene signature IL-8, CCL2/MCP-1, and SERP showed the best performance in distinguishing leprosy patients from healthy controls. These findings may have implications for developing a rapid blood-based test for early diagnosis of leprosy.
Assuntos
Hanseníase , Mycobacterium leprae , Antígenos de Bactérias , Biomarcadores , Diagnóstico Precoce , Humanos , Hanseníase/diagnóstico , Leucócitos Mononucleares , Mycobacterium leprae/genética , TranscriptomaRESUMO
Leprosy is an infectious disease caused by Mycobacterium leprae (M. leprae), with about 210,000 new cases per year worldwide. Although numerous risk loci have been uncovered by genome-wide association studies, the effects of common genetic variants are relatively modest. To identify possible new genetic locus involved in susceptibility to leprosy, whole exome sequencing was performed for 28 subjects including 14 patients and 12 unaffected members from 8 leprosy-affected families as well as another case and an unrelated control, and then the follow-up SNP genotyping of the candidate variants was studied in case-control sample sets. A rare missense variant in mitochondrial ribosomal protein S5 (MRPS5), rs200730619 (c. 95108402T>C [p. Tyr137Cys]) was identified and validated in 369 cases and 270 controls of Chinese descent (Padjusted = 0.006, odds ratio [OR] = 2.74) as a contributing factor to leprosy risk. Moreover, the mRNA level of MRPS5 was downregulated in M. leprae sonicate-stimulated peripheral blood mononuclear cells. Our results indicated that MRPS5 may be involved in leprosy pathogenesis. Further studies are needed to determine if defective MRPS5 could lead to impairment of energy metabolism of host immune cells, which could further cause defect in clearing M. leprae and increase susceptibility to infection.
Assuntos
Povo Asiático/genética , Predisposição Genética para Doença , Hanseníase/genética , Proteínas Mitocondriais/genética , Polimorfismo de Nucleotídeo Único , Proteínas Ribossômicas/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , China/epidemiologia , Feminino , Regulação da Expressão Gênica , Humanos , Hanseníase/epidemiologia , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
OBJECTIVE: To investigate the incidence of welder's pneumoconiosis in container manufacturing industries in Guangdong province and present some preventive and curative strategy. METHODS: Occupational epidemiology study methods were used to study the incidence of welder's pneumoconiosis in two container manufacturing enterprises (enterprise A and enterprise B). RESULTS: Before 2004, the rate of up-to-standard of weld fume concentration in the workplaces was relatively low (< or = 40%), and the maximum value of time weighted average (TWA) was 26.7 mg/m(3). After 2004, with the system of dust control, the rate of up-to-standard was 85% in the enterprise A. Of the 813 weld workers examined, 19 were diagnosed as welder's pneumoconiosis (stage I was 15, while I (+) was 4 ) and the incidence was 2.34% (19/813). The age of electric welders suffering from welder's pneumoconiosis and the duration of dust exposure were (33.45 + 4.64) and (8.04 + 1.97) years respectively. Chest radiographic examination showed mainly small round opacities "p". The value of MVV, FEF25%, FEF50% and FEF75% in lung function tests were significantly lower than those of the control group (P < 0.05 or P < 0.01). CONCLUSION: The main features of welder's pneumoconiosis in container manufacturing industries are the short duration of dust exposure, and small round opacities "p" in the chest radiographs. Therefore, it should be the key point of the prevention and treatment of occupational diseases in this industry to control the harmful weld fume.
Assuntos
Pneumoconiose/epidemiologia , Soldagem , Adulto , China/epidemiologia , Poeira , Feminino , Humanos , Masculino , Exposição Ocupacional/efeitos adversosRESUMO
OBJECTIVE: To study the hazards of gem dusts and the epidemiological features of the occurrence of gem worker's silicosis (GWS). METHODS: Epidemiological investigation in a gem factory in Guangdong province was carried out. RESULTS: The average dusts concentration in workshop was 2.3 mg/m(3), average free silica content was 94.64%. There were 47 patients diagnosed as silicosis. The prevalence of GWS was 4.58% (47/ 1,027). The age of patients suffering from silicosis and the duration of dust exposure were (27.04 +/- 3.25) years and (4.67 +/- 1.17) years respectively. Chest radiographic examination showed mainly small round opacities "q". The incidences of GWS complicated by pulmonary tuberculosis, spontaneous pneumothorax and pulmonary emphysema were 10.64%, 10.64%, 44.68% respectively. CONCLUSION: The hazards of gem dust to workers might be serious. Higher incidence, younger age suffered from silicosis, shorter duration of dust exposure, more complications, and poorer prognosis in gem workers appeared to be the main epidemiological features of GWS in Guangdong.
Assuntos
Doenças Profissionais/epidemiologia , Silicose/epidemiologia , Adulto , Poluentes Ocupacionais do Ar/intoxicação , China/epidemiologia , Feminino , Humanos , Masculino , Doenças Profissionais/etiologia , Silicose/etiologiaRESUMO
OBJECTIVE: To assay and study the microwave leakage of 4 types of interphones. METHODS: The radiation intensities of four types of 199 interphones were determined by the microwave leakage measure instrument of model ML-91 made in China. RESULTS: The average intensities of microwave leakage at a distance of 5 cm from aerial part and other parts of interphones during launching [(1 316.0 +/- 144.3), (971.0 +/- 131.6) microW/cm(2) respectively] were significantly higher than during waiting [(14.4 +/- 5.3), (13.2 +/- 4.9) microW/cm(2) respectively] (P < 0.01). The average intensities of microwave leakage at a distance of 50 cm from different parts were (357.3 +/- 27.8) microW/cm(2). The daily average intensity of microwave leakage to which the head, chest and abdomen exposed was (945.5 +/- 447.1) microW.h/cm(2) in total, that exceeded the hygienic standard of microwave in China (400 microW.h/cm(2)), during the normal communication by interphones. CONCLUSION: The microwave leakage was higher during launching than during waiting, and was the highest at the aerial part of the interphones. The microwave radiation of most interphones was higher than the current national standard. It may lead to potential effects on the owner of interphone, so protection against it should be made.
Assuntos
Telefone Celular , Micro-Ondas/efeitos adversos , Doses de RadiaçãoRESUMO
Leprosy is the disabling outcome of chronic infection with Mycobacterium leprae. The disease often evades early detection, particularly now that fewer clinicians are able to confidently diagnose the disease following the integration of leprosy control measures within general health services in many countries. Although leprosy is officially eliminated in China, endemic regions remain in some difficult-to-reach, underdeveloped areas in Southwest China. In order to better understand the extent of M. leprae infection and identify new leprosy cases in a timely manner, simple tools that can detect infection and the early disease are required. In this report we evaluated the performance of antigen-specific ELISA, the NDO-LID rapid diagnostic test, and antigen-specific whole blood assays (WBA) as potential diagnostic tools. Our data support the use of antibody detection tests and WBA to facilitate the diagnosis of multibacillary and paucibacillary leprosy, respectively. These tools could be invaluable for increased, but simplified, monitoring of individuals in order to provide referrals for clinical exam and early leprosy diagnosis.
Assuntos
Antígenos de Bactérias/sangue , Hanseníase/sangue , Mycobacterium leprae/isolamento & purificação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , China , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Hanseníase/genética , Hanseníase/patologia , Pessoa de Meia-Idade , Mycobacterium leprae/genética , Mycobacterium leprae/patogenicidadeRESUMO
The TaqMan real-time polymerase chain reaction (PCR) assay was evaluated systematically with respect to the standard curve, linear range, and used for detecting Mycobacterium leprae DNA in paraffin-embedded skin biopsy specimens from 60 confirmed leprosy patients and three healthy individuals and 29 other dermatoses and bacterial DNA from 21 different species. The test was further evaluated with 51 paucibacillary (PB) patients. The results showed that the test had good sensitivity (8 fg) and good specificity with no cross-reactivity with 21 other bacterial species and the control specimens, except one with Xanthomatosis. The real-time PCR detection rate for the 51 PB specimens was 74.5% (38 of 51). We conclude that the real-time PCR test is a useful adjunct test for diagnosing early stage or PB leprosy cases.
Assuntos
Hanseníase Paucibacilar/diagnóstico , Mycobacterium leprae/genética , Pele/microbiologia , Adolescente , Adulto , Idoso , Carga Bacteriana , Biópsia , Estudos de Casos e Controles , Criança , DNA Bacteriano/análise , Feminino , Humanos , Sequências Repetitivas Dispersas/genética , Hanseníase Paucibacilar/microbiologia , Masculino , Pessoa de Meia-Idade , Inclusão em Parafina , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e Especificidade , Dermatopatias/diagnóstico , Dermatopatias/microbiologia , Adulto JovemRESUMO
We evaluated the sensitivity and specificity of a nested-polymerase chain reaction (PCR) method for detection of Mycobacterium leprae DNA from whole blood. Whole-blood specimens were subjected to nested-PCR amplification of M. leprae repeat DNA sequences in 49 multibacillary (MB) and 30 paucibacillary (PB) leprosy patients, 96 household contacts (HHCs), 18 tuberculosis (TB) patients, and 35 normal healthy individuals. M. leprae DNA was detected in 95.92% (47/49) of MB, 70% (21/30) of PB, and 6.25% (6/96) of HHC, but it was not detected in 18 TB or 35 normal controls. The sensitivities of the anti-bovine serum albumin (ND-O-BSA) immunoglobulin M (IgM) and antifusion protein of ML0405-ML2331 IgG for MB were 97.96% and 89.8%, and these values for PB were 70% and 53.33%. However, the ND-O-BSA enzyme-linked immunosorbent assay (ELISA) had lower specificity, with relatively high false-positive results for TB patients (16.67%) and normal healthy controls (10%). Based on these promising findings, we propose the use of nested PCR of whole-blood samples along with ELISA test for early detection of leprosy cases.
Assuntos
DNA Bacteriano/sangue , Diagnóstico Precoce , Hanseníase/diagnóstico , Mycobacterium leprae/genética , Reação em Cadeia da Polimerase/métodos , Animais , Anticorpos Antibacterianos/sangue , Bovinos , China , DNA Bacteriano/isolamento & purificação , Ensaio de Imunoadsorção Enzimática , Humanos , Imunoglobulina M , Hanseníase/microbiologia , Mycobacterium leprae/isolamento & purificação , Sensibilidade e Especificidade , Soroalbumina Bovina/imunologiaRESUMO
OBJECTIVE: To explore the factors influencing the steady transmission of leprosy as indicated by new case detection rate in Qiubei county, Yunnan province, China despite the implementation of MDT for the last 25 years. METHODS: Information related to case-finding was collected. ELISA and PCR were applied to detect anti-PGL-1 antibody in sera and Mycobacterium leprae in nasal secretions respectively, in leprosy patients, their household contacts and the general population. M. leprae by PCR was also detected from water in the highly endemic villages. VNTR typing was performed to explore the mode and chain of transmission of M. leprae. RESULTS: Prior to 2001, the proportion of new cases detected from the examination of household contacts of leprosy patients was low (number, compared to), while the proportion of patients whose identification was delayed by more than 2 years, was high (number, compared to). Qualities of these two indicators has been improved, along with the improvement of leprosy control program since 2001, but the detection rates has been steady at 4-5/100 000 during 1986 - 2010. The PGL-1 seropositivity rate was 20% - 30% in general population, with the peak rate (30%) detected in the teenage population in the endemic villages. In addition to the fact that M. leprae was detected in nasal secretion from patients, their contacts and from water, the M. leprae VNTR genotypes were found to be highly similar between skin biopsy and nasal secretion in untreated cases. Families with multi-cases were clustered and located in the Northern part of the County, and the genotypes of M. leprae were identical within those families. The percentage of clusters was considerably higher in Northern rather than Southern parts of the County. CONCLUSION: Results from this molecular study demonstrated evidence that transmission of leprosy within the families and in the endemic-villages was severe. M. leprae were detected in waters from the endemic villages and others areas which might have a relation to the continued transmission of leprosy.
Assuntos
Hanseníase/transmissão , China/epidemiologia , Humanos , Hanseníase/epidemiologia , Epidemiologia Molecular , Prevalência , Microbiologia da Água , Poluentes da ÁguaRESUMO
OBJECTIVE: Multiple locus variable number-tandem repeat (VNTR) analysis (MLVA) had been proposed as a means of strain typing for tracking of source and studying the transmission chain of pathogens. However, empirical data for a defined population from scale and duration were lacking for studying the transmission chain of leprosy. METHODS: MLVA on 7 VNTR loci was applied to the strain typing on prevalent Mycobacterium leprae isolates collected from Qiubei county, Yunnan province during 2002-2006 in the study on the relationship between geographic distribution and genotypes of M. leprae. The strain typing, combined with conventional epidemiological investigation was performed to trace the transmission chain. RESULTS: (1) Phylogenetic analyses through application of PAUP 4.0, The M. leprae were grouped into A, B, C, D and E strains according to the allelic range 9, 11-13, 15-26 and > 26 on the GTA9 locus. The strains with 9 copies on GTA9 locus, was named A. (2) Genotypes of strains from the five multi-case families located at North and North-West parts were similar and belonged to A strains. VNTR patterns of intra-family were identical or similar but not identical inter-family. (3) Not only A cluster appeared higher proportion in total isolates but also distributes cluster, indicating ongoing transmission from recent findings. CONCLUSION: VNTR strain typing was suitable to trace the short chain of transmission in both small area and intra-families. Multi-case families might constitute epidemic foci and source of M. leprae in villages, causing the predominant strain or cluster which tends to be those identified in multi-case families and resulted in the spreading of leprosy. A long-term study was underway to reveal whether A strain was predominant strain and to observe the evolution of M. leprae in this spatially and temporally defined endemic population.
Assuntos
Genótipo , Hanseníase/microbiologia , Mycobacterium leprae/genética , Feminino , Humanos , Masculino , Repetições Minissatélites/genética , Epidemiologia Molecular , Mycobacterium leprae/classificação , Filogenia , Reação em Cadeia da PolimeraseRESUMO
OBJECTIVE: To understand the genotypic mapping of Mycobacterium leprae identified in China and to compare with those from other countries to select suitable alleles for epidemiological investigation in the transmission chain of leprosy. METHODS: Various number of tandem repeat(VNTR) in genomic DNA of Mycobacterium leprae was used in the present genotyping study. 33 skin biopsies from Wenshan prefecture,Yunnan province and 17 from other parts of China were studied. DNA extracted from skin biopsies of leprosy patients was subjected to PCR followed by agarose gel analysis and DNA sequencing to determine the number of repeats. RESULTS: Loci GGT-5,12-5,21-3 and 23-3 were as highly homogenous as 100%; The homogeneity of loci AC-8, 18-8, 27-5 and rpoT were 97%, 94%, 97% and 85% respectively. Loci GTA-9, AC-9 and 6-7 showed significant allelic diversity in isolates and the diversity of GTA-9 in Mycobacterium leprae isolated from China was also different from those identified other countries. We had subjected loci GTA-9 and the ten loci to phylogenetic tree analysis respectively. CONCLUSION: The present study revealed that the genotype of Mycobacterium leprae identified from China was close to the strains from the Philippines and India although a few loci were somehow differentiate. Locus 12-5 manifested as only 3 copies in China whereas 4-5 copies predominating in other countries. 12-5 locus might serve as a useful marker to diffrentiate Chinese strains from those in other countries. However, further study on the diversity of GTA-9 was needed in China. The molecular typing of Mycobacterium leprae from different geographic areas might be useful in studying the transmission of leprosy.