麻风病

983202 鼻分泌物及皮肤组织中麻风菌及其PGL-1抗原的检测/翁小满（北京热带医学研究所）…//临床皮肤科杂志.-1998,27（3）.-150～153 为了更好地理解麻风菌鼻携带在麻风病传播、维持中的作用,以及运用鼻携带检测来评价麻风病防治效果,比较了PCR和Dot-ELISA/ECL平行检测32例活动性麻风家内接触者鼻分泌物及皮肤组织中的麻风菌及其PGL-1抗原。结果显示,Dot-ELISA/ECL具有较好的敏感性、特异性,是一项适用于现场研究的简便、快速、经济的麻风流行病学工具。此外,用于免疫学试验,GVHP是一种吸附性好,适合于检测粘膜分泌物抗原的载体。图1表3参8 （原文摘要）983203 二甲胺四环素抗麻风杆菌活性及其治疗多菌型麻风的疗效/沈建平（中国协和医大皮研所）…//中华皮肤科杂志.-1998,31（3）.-189～190     

鼻分泌物及皮肤组织中麻风菌及共PGL—1抗原的检测

为了更好地理解麻风菌处携带在麻风病传播，维持中的作用，以及运用鼻携带检测来评价麻风病防治效果，比较了ＰＣＲ和Ｄｏｌ－ＥＬＩＳＡ／ＥＣＬ平行检测３２例活动性麻风患者，１３例愈后者和１４３名麻风家内接触者鼻分泌物及皮肤组织中的麻风菌及其ＰＧＬ－１抗原。     

麻风病

20052965麻风病免疫中的树突状细胞与T细胞的相互作用及其研究进展/翁小满(首都医大附属北京友谊医院北京热带医学研究所),胡永秀∥中国麻风皮肤病杂志.-2005,21(3).-200~203综述了有关麻风病在抗原递呈及诱导T细胞反应的研究;比较和分析DC递呈麻风菌及麻风菌组分抗原及其诱导不同的T细胞反应的机制;揭示了麻风菌是一个很独特的致病性分枝杆菌,它可能含有阻碍DC与T细胞相互作用的成分,而DC递呈麻风菌的组分抗原都可活化T细胞,这为逆转免疫无反应性的研究提供借鉴。参16(马宽玉)20052966酚糖酯-免疫球蛋白M和鼻分泌物中麻风菌PCR检测在麻风流行病学中的应用/翁小满(北京热带医学研究所),温艳,袁联潮…∥中国麻风皮肤病杂志.-2005,21(6).-425~428采用酚糖酯-酶联免疫吸附试验(PGL-ELISA)和检测鼻携带麻风菌的PCR方法,对流行乡村(云南省同烘和南丘)麻风病患者、家内接触者及普通人群麻风菌感染进行检测。结果:麻风病家内接触者的PGL-IgM阳性率和PCR检测的麻风菌携带率分别为30.4%和23.1%,两村普通村民PGL-IgM阳性率南丘村18.7%,同烘村20.86%,无...     

鼻拭子中麻风菌DNA的提取与检测

用鼻、咽喉、阴道等拭子收集分泌物标本,是检查病原微生物的一种简便的、非损伤性的方法.鼻腔是麻风菌进人人体和排除的通道,也是麻风病传播的传播途径之一.鼻拭子中麻风菌DNA检测,是麻风病流行病学研究的重要工具之一.但是,受鼻拭子取量少或携带菌量少的限制,传统的酚、氯仿抽提DNA,显然不适宜.本文介绍三种DNA提取方法,比较其优缺点及克服的方法.     

斑点-酶联免疫吸附试验检测麻风病人尿和鼻分泌物中的麻风杆菌特异性抗原

作者应用斑点-酶联免疫吸附试验(Dot-ELISA)对102例(TT30例、BT22例、BB5例、BL20例、LL25例)新发未治麻风病人尿及鼻分泌物中麻风杆菌特异性抗原进行了检测。这些病人的病期为1月～13年。每个病人取50ml尿液,从中提取脂质后溶于已烷备用。另外以湿棉花拭子清洗病人鼻腔,将清洗物混悬于5 ml生理盐水中,用真空冷冻干燥法浓缩20倍,用于抗原检测。以Dot-ELISA检测时,使用2种第一抗体,一种为从LL型病人中制取的抗麻风超免疫混合血清,并在使用前用包括卡介苗在内的6种可培养分支杆菌可溶性抗原吸收,其工     

麻风病血清流行病学调查与化学预防的实施

目的:评价化学预防的保护性,本研究依据血清流行病学调查,对两个麻风病高流行村麻风病接触者和一般人群实施利福平化学预防.方法:采用临床查体、检测血清酚糖脂-1(PGL-Ⅰ)抗体和鼻分泌物麻风菌,了解人群的麻风菌感染状况,为评价预防治疗效果提供科学依据.结果:YG村临床普查率达98%,发现早期病人2例;接触者血清PGL-Ⅰ抗体阳性率76%,麻风菌鼻携带率35%,预防服药率达98%.HG村临床普查率91%,发现早期病人1例,人群血清PGL-Ⅰ抗体受检率54%,其中血清PGL-Ⅰ抗体阳性率33%,服药率达85%.结论:对亚临床感染率较高的人群实施化学预防很有必要.预防后的发病率与血清学等实验室数据可为评价预防治疗提供依据.     

酚糖酯-免疫球蛋白M和鼻分泌物中麻风菌PCR检测在麻风流行病学中的应用

目的通过对流行乡村(同烘和南丘)麻风病患者、家内接触者及普通人群麻风菌感染的检测,评估实验流行病学对预测麻风病传播的意义。方法采用酚糖酯-酶联免疫吸附试验(PGL-ELISA)和检测鼻携带麻风菌的PCR方法,开展流行病学调查。结果(1)麻风病家内接触者的酚糖酯-免疫球蛋白M(PGL-IgM)阳性率和PCR检测的麻风菌鼻携带率分别为30.4%和23.1%;但PGL抗体阳性率在家内接触者和普通村民之间却无显著性差异。(2)两村普通村民的PGL-IgM阳性率,在统计学上无显著差异。然而,在<20岁的年龄组中,同烘村的PGL-IgM阳性率却显著高于南丘村。无论同烘或南丘村,PGL-IgM阳性率高峰均在<20岁的年龄组。随年龄的增加,阳性率逐渐下降。此外,女性的PGL-IgM阳性率高于男性。结论两村的新发现病人主要为年轻人,这与两村PGL-IgM阳性高峰位于<20岁年龄组相关。在<20岁的年龄组中,同烘村的PGL-IgM阳性率显著高于南丘村,除与同烘村患病率和发现率均高于南丘村相关,也与消除麻风病运动(LEC)后,同烘村仍有新病人出现有关。这一现象似乎支持麻风患病率与PGL-IgM阳性率相关。为评价麻风病的传播是否得到控制,以PGL的血清学仍是一种有用的方法。     

麻风病免疫中的树突状细胞与T细胞的相互作用及其研究进展

研究瘤型麻风病的特异性免疫无反应性及其逆转机制，是研制麻风疫苗和发展免疫治疗的基础。单核细胞来源的树突状细胞(DC)与T细胞共同培养，是研究DC与T细胞相互作用的理想模式。本文综述了有关麻风病在抗原递呈及诱导T细胞反应的研究；比较和分析DC递呈麻风完整菌及麻风菌组分抗原及其诱导不同的T细胞反应的机制；揭示了麻风菌是一个很独特的致病性分枝杆菌，它可能含有阻碍DC与T细胞相互作用的成分，而DC递呈麻风菌的组分抗原却可活化T细胞这为逆转免疫无反应性的研究提供借鉴。     

应用ROC曲线评价不同抗原检测麻风病特异性抗体的价值

目的应用ROC曲线评价不同抗原采用ELISA检测麻风病特异性抗体的价值。方法筛选经金标准确认的61例麻风病和42例对照标本,应用ELISA法检测,比较不同的麻风特异性抗原的检测效率。结果 LID-1,NDO-BSA,NDO-LID三种抗原检测多菌型麻风(MB)患者抗体阳性率分别为92.11%,89.47%,92.11%;检测少菌型麻风(PB)患者抗体阳性率分别为52.17%,60.87%,47.83%。将病例组根据不同的型别分为MB和PB,MB,PB三个组,通过ROC曲线计算ROC曲线下面积,当病例组包括MB和PB时,NDO-LID抗原AUROC面积最大,等于0.86;当病例组只包括MB时,也是NDO-LID抗原AUROC面积最大,等于0.95,当病例组为PB时,NDO-BSA抗原AUROC面积最大,等于0.74。提示ELISA方法对MB检测效率高于PB,三种抗原中检测MB患者,NDO-LID最优,对PB患者NDO-BSA稍优于其他抗原。结论 ROC曲线是评价不同抗原检测麻风病特异性抗体效率的有效方法。     

麻风病

20070660 多菌型麻风患5种自身抗体检测结果分析;20070661 1952-2005年368例麻风死亡原因分析；20070662门诊一日遇二例麻风患；20070663 一本全面、新颖和实用的麻风病专——《现代麻风病学》；20070664 麻风病误诊1例；20070665 汕头市澄海区1991—2005年麻风病流行病学分析；[编按]     

Detection of a Mycobacterium leprae cell wall antigen in the urine of untreated and treated patients.

A total of 90 leprosy patients, 12 household contacts and 10 normal subjects were studied for the detection of Mycobacterium leprae cell wall antigen in urine using monoclonal antibody (ML30A2 IgG). In untreated multibacillary leprosy (BL-LL) the M. leprae cell wall antigen could be demonstrated in the urine of 14 (64%) patients by immunofluorescence (IF) and 22 (100%) by ELISA. In untreated paucibacillary leprosy (TT-BT), it could be demonstrated in 3 (11.5%) and in 13 (50%) patients by IF and ELISA methods respectively. All but 1 household contact (later confirmed to have BL leprosy) and all 10 normal subjects' urine was negative for M. leprae cell wall antigen by both methods. The same antigen was, however, demonstrated in urine of 50% paucibacillary patients who had received 6 months of treatment and in 68% multibacillary patients who had received 24 months of WHO recommended multidrug therapy. M. leprae cell wall antigen assays in urine will not be useful in the follow-up of leprosy patients on multidrug therapy.     

Immunogenicity of Mycobacterium leprae unique antigens in leprosy endemic populations in Asia and Africa

Ongoing transmission of leprosy is evident from the stable disease incidence in high burden areas. Tools for early detection of Mycobacterium leprae (M. leprae) infection, particularly in sub-clinically infected individuals, are urgently required to reduce transmission. Following the sequencing of the M. leprae genome, many M. leprae-unique candidate proteins have been identified, several of which have been tested for induction of M. leprae specific T cell responses in different leprosy endemic areas. In this study, 21 M. leprae-unique proteins and 10 peptide pools covering the complete sequence of five M. leprae-unique proteins (ML0576, ML1989, ML1990, ML2283, and ML2567) were evaluated in 160 individuals in Nepal and Ethiopia. These included: tuberculoid and borderline tuberculoid (TT/BT), borderline borderline and borderline lepromatous (BB/BL) leprosy patients; healthy household contacts (HHC); tuberculosis (TB) patients and endemic controls (EC). Immunogenicity of the proteins was determined by IFN-gamma secretion via stimulation of PBMC in 6 days lymphocyte stimulation tests (LST) or in whole blood assays (WBA). In LST, BB/BL patients (40%) responded to ML0573 and ML1601 whereas ML1604 was most immunogenic in TT/BT (35%) and HHC (36%). Additionally, significant numbers of EC displayed IFN-gamma production in response to ML0573 (54%), ML1601 (50%) and ML1604 (54%). TB patients on the other hand, hardly responded to any of the proteins except for ML1989. Comparison of IFN-gamma responses to ML0121, ML0141 and ML0188 for TT/BT patients showed specific increase in diluted 6 days WBA compared to the undiluted 24 hours WBA, whereas EC showed a reduced response in the diluted WBA, which may indicate detection of disease-specific responses in the 6 days WBA. In summary, identification of multiple M. leprae proteins inducing M. leprae-specific T cell responses in groups at high risk of developing leprosy may contribute to improve early detection for M. leprae infection.     

Use of non-conventional antigen: M. habana in detecting M. leprae antibodies from leprosy patients and contacts in FLA-ABS test

An indirect immunofluorescent (FLA-ABS) test has been developed to detect M. leprae specific antibodies in the active and subclinical cases of leprosy. An antigenically related mycobacterium, M. habana, was used as an antigen to detect M. leprae specific antibodies in the sera samples of leprosy patients. A comparison was made with M. leprae antigen using same set of sera samples. M. habana is capable of detecting anti-M. leprae antibodies in the serum samples of leprosy patients, previously absorbed with various mycobacterial antigens, cardiolipin and lecithin, almost to the same percentage as M. leprae. Possible use of M. habana antigen as an alternative to M. leprae, in the serodiagnosis of leprosy, has been discussed.     

Detection of lepromatous leprosy patients shedding M. leprae in nasal droppings by enzyme immunoassay

Enzyme immunoassays (EIAs) for detection of lepromatous leprosy (LL) patients harbouring M. leprae in nasal mucosa are described. One EIA measures IgM antibodies against the synthetic disaccharide (ND-BSA) residue of phenolic glycolipid I of M. leprae, whereas the other titrates primarily IgG antibodies against sonicate supernatant antigens of Mycobacterium w. (M.w.). Fifty coded leprosy sera were analysed by EIAs under a double blind code. Amongst the 20 LL patients with positive nasal smear, 18 (90%) were positive in EIA based on ND-BSA, in comparison to 19 (95%) in EIA using M.w. antigens. The assays can be performed on fresh serum samples or on blood samples collected on filter paper discs. These assays can be useful for leprosy control programmes.     

Study of cytokine response against panel of purified Mycobacterium leprae antigens by using whole blood assay in subjects residing in a resettlement village of cured leprosy patients

Mycobacterium leprae being an intracellular pathogen, cell mediated immunity is very important in the clinical outcome of leprosy. Manifestation of the disease is correlated with the level and type of cell mediated immune response. The main objective of this study was to analyse TNF-alpha and IFN-gamma production by T-cells when challenged with different M. leprae purified antigens in subjects with known exposure. 50 subjects residing in resettlement village of cured leprosy patients were included in the study. Whole Blood assay studies were undertaken in which the blood was placed in culture and was challenged with 35-kDa antigen, whole M. leprae cells, M. leprae cell wall antigen and M. leprae soluble antigen minus LAM. T-cell derived cytokines TNF-alpha and IFN-gamma were measured by ELISA. It was observed that challenging the lymphocytes with 35-kDa antigen, the cell wall antigen and M. leprae soluble antigen minus LAM resulted in increased levels of IFN-gamma whereas challenge with 35-kDa antigen and M. leprae cell wall antigen resulted in increased levels of TNF-alpha.     

Leprosy: review of the epidemiological,clinical, and etiopathogenic aspects - Part 1

Leprosy is caused by Mycobacterium leprae and has been known since biblical times. It is still endemic in many regions of the world and a public health problem in Brazil. The prevalence rate in 2011 reached 1.54 cases per 10,000 inhabitants in Brazil. The mechanism of transmission of leprosy consists of prolonged close contact between susceptible and genetically predisposed individuals and untreated multibacillary patients. Transmission occurs through inhalation of bacilli present in upper airway secretion. The nasal mucosa is the main entry or exit route of M. leprae. The deeper understanding of the structural and biological characteristics of M. leprae, the sequencing of its genome, along with the advances in understanding the mechanisms of host immune response against the bacilli, dependent on genetic susceptibility, have contributed to the understanding of the pathogenesis, variations in the clinical characteristics, and progression of the disease. This article aims to update dermatologist on epidemiological, clinical, and etiopathogenic leprosy aspects.     

Comparison of PCR mediated amplification of DNA and the classical methods for detection of Mycobacterium leprae in different types of clinical samples in leprosy patients and contacts

Traditional staining and microscopic examination techniques for the detection of Mycobacterium leprae, DNA amplification by polymerase chain reaction (PCR) of a 531-bp fragment of the M. leprae specific gene encoding the 36-kDa antigen, and serodiagnosis with M. leprae specific antigens (PGL-1 and D-BSA) were compared on different clinical specimens (serum samples, slit-skin smears, biopsies and swabs) from 60 leprosy patients attending the Sanatorium of Fontilles. Patients were divided into groups; (i) 20 multibacillary patients (MB) with positive bacteriological index (BI) by conventional methods and on WHO multidrug therapy (MDT); (ii) 30 MB patients with negative BI and completed minimum 2 years treatment MDT; (iii) 10 paucibacillary (PB) patients who had completed 6 months MDT at least 8 years ago. Control groups included four non-leprosy patients for PCR methods and 40 health control patients and 10 tuberculosis patients for serological methods. In the multibacillary BI positive group, there was a good correlation between all methods. All tests were negative in the paucibacillary group, although only a few patients were tested and all had been treated many years ago. One must be cautious concerning the diagnostic potential of these techniques in this type of leprosy. We also studied different combinations of leprosy diagnosis methods to determine the potential risk in a leprosy contact individuals group. The prevalence of antibodies to M. leprae antigens in serum was measured, together with the presence of M. leprae DNA in the nose and lepromin status in a group of 43 contacts of leprosy patients (12 household and 31 occupational) to evaluate the maintenance of infection reservoirs and transmission of the disease. Only two individuals were found to form a potential high risk group.