Determinação do agente microbiano, perfil de resistência antimicrobiana e produção de biofilme em úlceras de membros inferiores em pacientes com sequelas de hanseníase e outras doenças crônicas / Determination of the microbial agent, antimicrobial resistance profile and biofilm production in lower limb ulcers in patients with leprosy sequelae and other chronic diseases

Úlceras crônicas são definidas quando o processo de reparação do tecido excede o período de 3 meses, dificultando sua cicatrização. Sua etiologia pode ser multifatorial, como a ocorrência de traumas e consequência de patologias, como hanseníase, hipertensão e diabetes. As úlceras abrigam diversos microrganismos colonizadores e residentes que podem tornar-se potenciais agravantes a sua condição clínica, visto sua capacidade de formação de biofilmes e resistência antimicrobiana, diminuindo a eficácia da terapêutica. O objetivo deste trabalho foi determinar os agentes microbianos presentes em úlceras de pacientes com doenças crônicas atendidos no ambulatório de feridas do Instituto Lauro de Souza Lima, avaliar a susceptibilidade antimicrobiana destes isolados e sua capacidade de produção de biofilme, bem como comparar os resultados evidenciados por swab e biópsia e correlacionar os resultados microbiológicos com dados clínicos dos pacientes. Foram coletadas amostras de exsudato por swab e biópsia de úlceras crônicas dos participantes com doenças crônicas. As amostras foram semeadas em ágar sangue, manitol, cetrimide e MacCnkey para posterior identificação microbiana. Também foi desempenhada a determinação da susceptibilidade aos antimicrobianos e capacidade de produção de biofilme dos isolados identificados por swab e biópsia. Foram identificados 47 microrganismos no total, sendo 26 (55%) isolados presentes no swab e 21 (45%) em biópsia. P. aeruginosa, P. mirabilis e S. aureus foram as bactérias comumente prevalentes em ambos os materiais de coleta, com predomínio de P. aeruginosa. Apenas 16 (36%) das bactérias demonstraram capacidade de produzir biofilme, com destaque para o grupo dos gram-positivos (92%) que também exibiram alto perfil de susceptibilidade frente linezolida e vancomicina. Meropenem foi o único fármaco a mostrar eficácia frente as cepas de P. aeruginosa presentes, enquanto o grupo das enterobactérias apresentaram menor resposta frente a amoxicilina com ácido clavulânico. Swab e biópsia apresentaram uma concordância geral de 60%, semelhante ao observado por outros estudos. Tais diferenças podem se dar devido à presença de colonizadores. A cobertura de zinco e bota de Unna foi correlacionada à ausência de sinais flogísticos de infecção. Os dados sociodemográficos mostram prevalência de indíviduos com baixa escolaridade e idade acima de 60 anos. O swab é menos invasivo e mais utilizado devido sua facilidade e baixo custo em relação a biópsia; contudo, deve ser considerado com mais cautela na análise dos resultados microbiológicos.

Chronic wounds are defined when the tissue repair process exceeds the period of 3 months, making it difficult to heal. Its etiology can be multifactorial, such as the occurrence of trauma and consequences of pathologies, such as leprosy, hypertension, and diabetes mellitus. Ulcers harbor several colonizing and resident microorganisms that can become potential aggravating factors for their clinical condition, given their ability to form biofilms and their antimicrobial resistance, decreasing the therapeutic efficacy. This study aimed to determine the microbial agents present in ulcers of patients with chronic conditions treated at the wound clinic of the Instituto Lauro de Souza Lima, to evaluate their antimicrobial susceptibility and ability to produce biofilm, as well as to compare the results evidenced by swab and biopsy and correlate the microbiological results with clinical data of the patients. Exudate samples were collected by swab and biopsy of leg ulcers from participants with chronic diseases. Samples were seeded on sheep blood agar, mannitol, cetrimide and MacConkey agar for subsequent microbial identification. The determination of antimicrobial susceptibility and biofilm production capacity of isolates identified by swab and biopsy was also performed. A total of 47 microorganisms were identified, 26 (55%) of which were isolated from the swab and 21 (45%) from the biopsy. P. aeruginosa, P. mirabilis and S. aureus were the commonly prevalent bacteria in both collection materials, with predominance of P. aeruginosa. Only 16 (36%) bacteria demonstrated the ability to produce biofilm, with emphasis on the gram-positive group (92%) that also exhibited a high profile of susceptibility to linezolid and vancomycin. Meropenem was the only drug to show efficacy against the strains of P. aeruginosa present, while the group of enterobacteria showed less response against amoxicillin with clavulanic acid. Swab and biopsy showed an overall agreement of 60%, similar to that observed by other studies. Such differences may occur due to the presence of colonizers. Zinc coating and Unna boot correlated with the absence of phlogistic signs of infection. Sociodemographic data show a prevalence of individuals with low education and aged over 60 years. The swab is less invasive and more used due to its ease and low cost compared to biopsy; however, it should be considered with more caution in the analysis of microbiological results

Resistencia a los medicamentos del Mycobacterium Leprae detectados en cultivo de almohadilla plantar de ratón entre 1997 y 2013 en Malasia / No disponible

Antecedentes: Después de tres décadas de implementación de la multiterapia (MDT), consistente en una combinación de rifampicina, dapsona y clofazimina, en Malasia la aparición de resistencia farmacológica del Mycobacterium leprae constituye una preocupación, ya que puede llevar al fracaso del tratamiento y la recidiva de la enfermedad. Objetivos: Determinar el modelo de resistencia farmacológica del M. leprae en Malasia. Métodos: Se analizaron los cultivos en almohadilla plantar de ratón (MFP) de todas las biopsias cutáneas de pacientes con lepra borderline lepromatosa y lepra lepromatosa enviados a la Unidad de la Lepra, Laboratorio Nacional de Salud Publica, Sungai Buloh, Malasia, entre 1997-2013. Resultados: Se realizaron 651 cultivos MFP. La edad media de los pacientes fue de 41 anos (rango: 6-88). La proporción varón/hembra era de 3·8:1. Cuatrocientos cuarenta y cuatro pacientes (69·1%) eran malayos. La proporción de M. leprae positivo en cultivo era del 66·6% (433 of 651). El Índice Bacteriologico (IB) y el Índice Morfológico (IM) promedios para los cultivos positivos fue de 3·7 and 2·8 respectivamente. El IB y el IM de los que no crecieron en la MFP eran significativamente menores que los que presentaban cultivos positivos (P < 0·001). La dapsona presento el mayor índice de resistencia del 55% (238 of 433). Sin embargo, el elevado grado de resistencia a la dapsona (0·01%) fue de 6·24%. Hubo 407 MFP con rifampicina 0·003% y 12 (2·9%) resultaron resistentes a la misma. La clofazimina presento el menor grado de resistencia intermedia (0·001%) que fue del 0·2% (1 of 429). No había diferencias significativas entre el patrón de resistencia y género o nacionalidad de los pacientes. Conclusiones: Mas de la mitad de los cultivos MFP presentaron resistencia de baja intensidad a la dapsona; menos del 3% eran resistentes a la rifampicina y la resistencia a la clofazimina resulto muy baja

Background: After three decades of implementing multidrug therapy (MDT) consisting of rifampicin, dapsone and clofazimine in Malaysia, the drug resistance pattern of Mycobacterium leprae is a growing concern as it may lead to failure of treatment and relapse of disease. Objective: To determine the drug resistance patterns of M. leprae in Malaysia. Methods: Mouse footpad (MFP) culture of all skin biopsy samples from patients with borderline lepromatous and lepromatous leprosy sent to the Leprosy Unit, National Public Health Laboratory, Sungai Buloh, Malaysia between 1997-2013 were retrospectively studied. Results: There were 651 MFP cultures performed. The mean age of patients was 41 years old (range: 6-88). The male: female ratio was 3·8:1. Four hundred and forty four patients (69·1%) were Malaysian. The rate of positive M. leprae culture was 66·6% (433 of 651). The mean Bacteriological Index (BI) and median Morphological Index (MI) for those with positive culture were 3·7 and 2·8 respectively. The mean BI and MI of those which failed to grow in the MFP were significantly lower than those with positive cultures (P < 0·001). Dapsone has the highest resistance rate of 55% (238 of 433). Nevertheless, high degree dapsone resistance (0·01%) was 6·24%. There were 407 MFP tests using rifampicin 0·003% and 12 (2·9%) were resistant to it. Clofazimine has the lowest intermediate degree (0·001%) resistance rate of 0·2% (1 of 429). There were no significant differences between the drug resistance pattern and the gender or the nationality of the patients. Conclusion: More than half of our positive MFP cultures showed low-level resistance to dapsone; less than 3% were resistant to rifampicin, and clofazimine resistance remained very low

Métodos de determinación de sensibilidad a los antimicrobianos en micobacterias / Methods for determining the antimicrobial susceptibility of mycobacteria

Las micobacterias son un amplio grupo de microorganismos en el que múltiples especies son causa de una importante morbimortalidad, como la tuberculosis y la lepra. La aparición y diseminación de cepas del complejo Mycobacterium tuberculosis resistentes a diversos fármacos constituye en la actualidad uno de los problemas sanitarios de mayor gravedad a nivel mundial. Por otro lado, las micobacterias diferentes de M. tuberculosis y Mycobacterium leprae, denominadas micobacterias no tuberculosas (MNT), son aislamientos cada vez más frecuentes, requiriendo en muchos casos un tratamiento que precisa una orientación sobre la sensibilidad de estos microorganismos a los antimicrobianos. En el presente artículo se revisan los métodos para determinar la sensibilidad in vitro a los antimicobacterianos de los aislamientos del complejo M. tuberculosis y las MNT más relevantes. Además, también se realiza un análisis de las técnicas moleculares de detección rápida de la resistencia a partir de las muestras clínicas (AU)

Mycobacteria are a large group of microorganisms, multiple species of which are major causes of morbidity and mortality, such as tuberculosis and leprosy. At present, the emergence and spread of multidrug-resistant strains of Mycobacterium tuberculosis complex are one of the most serious health problems worldwide. Furthermore, in contrast to M. tuberculosis and Mycobacterium leprae, non-tuberculous mycobacteria (NTM) are more frequently isolated and, in many cases, treatment is based on drug susceptibility testing. This article is a review of the different methods to determine the in vitro drug susceptibility of M. tuberculosis complex and the most relevant NTM isolates. The molecular techniques currently used for rapid detection of resistance of clinical specimens are also analysed (AU)

Methods for determining the antimicrobial susceptibility of mycobacteria. / Métodos de determinación de sensibilidad a los antimicrobianos en micobacterias.

Mycobacteria are a large group of microorganisms, multiple species of which are major causes of morbidity and mortality, such as tuberculosis and leprosy. At present, the emergence and spread of multidrug-resistant strains of Mycobacterium tuberculosis complex are one of the most serious health problems worldwide. Furthermore, in contrast to M. tuberculosis and Mycobacterium leprae, non-tuberculous mycobacteria (NTM) are more frequently isolated and, in many cases, treatment is based on drug susceptibility testing. This article is a review of the different methods to determine the in vitro drug susceptibility of M. tuberculosis complex and the most relevant NTM isolates. The molecular techniques currently used for rapid detection of resistance of clinical specimens are also analysed.

Transmission of Drug-Resistant Leprosy in Guinea-Conakry Detected Using Molecular Epidemiological Approaches.

Molecular drug susceptibility testing was performed on skin biopsies from 24 leprosy patients from Guinea-Conakry for the first time. We identified primary drug resistance in 4 cases and a dapsone-resistant cluster caused by the same strain. Primary transmission of drug-resistant Mycobacterium leprae, including a rifampicin-resistant strain, is reported.

Prevalence and antibiotic susceptibility of Mycoplasma hominis and Ureaplasma urealyticum in genital samples collected over 6 years at a Serbian university hospital.

BACKGROUND: Mycoplasma hominis and Ureaplasma urealyticum are implicated in a wide array of infectious diseases in adults and children. Since some species have innate or acquired resistance to certain types of antibiotics, antibiotic susceptibility testing of mycoplasma isolated from the urogenital tract assumes increasing importance. AIMS: To evaluate the prevalence and antibiotic susceptibility of M. hominis and U. urealyticum in genital samples collected between 2007 and 2012. METHODS: Three hundred and seventy three patients presenting with symptoms of sexually transmitted diseases, infertility or risky sexual behaviour, who had not taken antibiotics in the previous 6 weeks and had ≥10 WBC per high power field on genital smears were studied. Urethral samples were taken in men and endocervical samples in women. The mycoplasma IST-2 kit was used for organism identification and for testing susceptibility to doxycycline, josamycin, ofloxacin, erythromycin, tetracycline, ciprofloxacin, azithromycin, clarithromycin and pristinamycin. RESULTS: U. urealyticum was isolated from 42 patients and M. hominis from 11 patients. From 9.8% of isolates, both organisms were grown. All M. hominis isolates were resistant to tetracycline, clarithromycin and erythromycin while U. urealyticum was highly resistant to clarithromycin (94.6%), tetracycline (86.5%), ciprofloxacin (83.8%) and erythromycin (83.8%). M. hominis was sensitive to doxycycline (83.3%) and ofloxacin (66.7%) while most U. urealyticum strains were sensitive to doxycycline (94.6%). LIMITATIONS: Inability of the commercial kit used in the study to detect other potentially pathogenic urogenital mycoplasmas (Ureaplasma parvum, Mycoplasma genitalium). CONCLUSION: There is significant resistance of U. urealyticum and M. hominis to tetracycline and macrolides. The most active tetracycline for genital mycoplasmas was found to be doxycycline, which continues to be the drug of first choice.

Prevalência e perfil de susceptibilidade a antimicrobianos dos uropatógenos em pacientes atendidos no Instituto Lauro de Souza Lima/Bauru/SP / Prevalence and antimicrobial susceptibility profile of uropathogens in patients treated at the Instituto Lauro de Souza Lima / Bauru / SP

As infecções de trato urinário (ITU) estão entre as doenças infecciosas mais comuns na prática clínica. O objetivo do presente estudo foi investigar a prevalência de microrganismos patogênicos, analisando a faixa etária e gênero mais acometidos bem como o perfil de resistência aos antimicrobianos. O estudo caracterizou-se por uma pesquisa histórica documental, na qual se analisou registros de 605 uroculturas realizadas pelo Laboratório de microbiologia do Instituto Lauro de Souza Lima, na cidade de Bauru/SP, no período de Outubro de 2010 a Outubro de 2015. Foram incluídos pacientes de ambos os gêneros e de todas as idades. Os dados foram coletados a partir do livro de registro de exames do laboratório e do sistema de informatização SIGH e transcritos para uma planilha eletrônica utilizando o programa Microsoft Office Excel. Das 605 uroculturas analisadas, 24,2% apresentaram resultados positivos para ITU. Dentre as positivas, 75,3% foram de pacientes do gênerofeminino. Analisando a incidência dos microrganismos, a bactéria Escherichia coli foi a mais isolada, apresentando maior resistência à Penicilina G, quinolonas e trimetoprim-sulfamatoxazol. Pode-se concluir que o correto diagnóstico é imprescindívelna escolha e na instituição da antibioticoterapia mais adequada, evitando o uso indiscriminado de antimicrobianos...

The draft genome of Mycobacterium aurum, a potential model organism for investigating drugs against Mycobacterium tuberculosis and Mycobacterium leprae.

Mycobacterium aurum (M. aurum) is an environmental mycobacteria that has previously been used in studies of anti-mycobacterial drugs due to its fast growth rate and low pathogenicity. The M. aurum genome has been sequenced and assembled into 46 contigs, with a total length of 6.02Mb containing 5684 annotated protein-coding genes. A phylogenetic analysis using whole genome alignments positioned M. aurum close to Mycobacterium vaccae and Mycobacterium vanbaalenii, within a clade related to fast-growing mycobacteria. Large-scale genomic rearrangements were identified by comparing the M. aurum genome to those of Mycobacterium tuberculosis and Mycobacterium leprae. M. aurum orthologous genes implicated in resistance to anti-tuberculosis drugs in M. tuberculosis were observed. The sequence identity at the DNA level varied from 68.6% for pncA (pyrazinamide drug-related) to 96.2% for rrs (streptomycin, capreomycin). We observed two homologous genes encoding the catalase-peroxidase enzyme (katG) that is associated with resistance to isoniazid. Similarly, two embB homologues were identified in the M. aurum genome. In addition to describing for the first time the genome of M. aurum, this work provides a resource to aid the use of M. aurum in studies to develop improved drugs for the pathogenic mycobacteria M. tuberculosis and M. leprae.

Patrones de resistencia farmacológica en muestras de Mycobacterium leprae de pacientes de lepra con recidivas en los Hospitales de the Leprosy mission (TLM) de India / No disponible

La implementación de la multiterapia (MDT) en los programas de control de la lepra ha reducido la prevalencia global de la enfermedad durante las dos últimas décadas. Después de muchos años de administrar la MDT hay que evaluar si pueden haber casos de resistencia del Mycobacterium leprae frente a la multiterapia. Esto constituye una gran preocupación, especialmente durante la actual fase de eliminación de la lepra. En este trabajo se obtuvieron frotis cutáneos de 140 casos de lepra con recidivas de distintos hospitales de The Leprosy Mission en la India. El DNA extraído de 111 (79%) de estas muestras se analizó para el análisis de genes asociados a la resistencia por M. leprae. Más del 90% de los pacientes recidivaron como multibacilares (MB). En nuestro estudio, cuatro de las muestras de DNA (3·6%) revelaron mutaciones asociadas con resistencia a la rifampicina. También se detectó que las mutaciones asociadas con la resistencia a la dapsona u ofloxacino se detectaron en 9 (8·1%) de las muestras; dos muestras presentaban resistencia a ambos. Se requiere más vigilancia e intervenciones adecuadas para asegurar la eficacia futura de la quimioterapia en la lepra

Implementation of multidrug therapy (MDT) in leprosy control programmes has significantly reduced the global prevalence of the disease in the last two decades. After many years of use of MDT, it is expected that drug resistance in Mycobacterium leprae may emerge. This is a major concern, especially during the stage of elimination. In the present study, slit-skin smears were collected from 140 leprosy relapse cases from different Leprosy Mission hospitals across India. DNA extracted from 111 (79%) of these samples was analysed for the genes associated with drug resistance in M. leprae. More than 90% of the patients relapsed as multibacillary (MB) cases. In our study, four (3·6%) of the DNA samples analysed showed mutations associated with rifampicin resistance. We also observed that mutations associated with resistance to dapsone and ofloxacin were observed in 9 (8·1%) of the DNA samples each; two samples had both dapsone and ofloxacin resistance. Further surveillance and appropriate interventions are needed to ensure the continued success of chemotherapy for leprosy

[What Hansen's disease research learned from tuberculosis research: from molecular biological aspect].

As for the Mycobacterium leprae which is a causative agent of Hansen's disease, many studies had been done since it was identified in 1873. However, those studies, at the same time, experienced many struggles because of the difficulty of culture of M. leprae on the artificial growth media. Hence, the study of Hansen's disease progressed by taking the knowledge from the study of tuberculosis caused by the bacteria belonging to the same genus, genus Mycobacterium. For instance, the knowledge of mutations in specific genes responsible for rifampicin- and quinolone-resistance in M. tuberculosis led the elucidation of drug-resistant acquisition mechanism of M. leprae. Similarly, it is necessary for the researcher of Hansen's disease to get important information from the latest topic of the tuberculosis study and utilize them to the study of the disease.

Rifampicin-resistant Mycobacterium leprae in an elderly leprosy patient in the people's Republic of China.

The reported number of registered leprosy patients worldwide declined with the introduction of multidrug therapy. However, the emergence of rifampicin resistance in leprosy patients engenders difficulties for an individual patient, and its dissemination could pose a threat to leprosy control. This study reports an elderly patient who was diagnosed with rifampicin-resistant lepromatous leprosy. This case indicates that inadequate treatment and poor compliance may eventually result in rifampicin resistance in Mycobacterium leprae and clinical relapse.

Clinical analysis of multibacillary leprosy patients after 1-year fixed World Health Organization recommended multidrug therapy at Yangon General Hospital, Myanmar.

This study included 200 randomly selected multibacillary leprosy cases who had completed 1 year of fixed World Health Organization recommended multidrug therapy (WHO-MDT) without prior dapsone (DDS) monotherapy. The time interval after release from treatment varied from a few months to 8 years. All cases were clinically reviewed in 2006 by comparison with their old clinical records. Reactions, particularly reversal reactions, occurred frequently among patients who had completed MDT within the last 3 years. It was difficult to distinguish relapse cases and late reversal reactions in skin smear-negative multibacillary cases. Based on bacteriological and histological analyses, one patient was confirmed to have relapsed 1 year after release from treatment. The overall relapse rate was 0.5%. No drug resistance mutations were detected by polymerase chain reaction or dot blot hybridization. The present study indicates that it is important to follow up patients for several years after completion of MDT in order to detect possible lepra reactions and relapses.

[Study of rifampin and dapsone resistance in three patients with recurring leprosy]. / Estudio de resistencia a la rifampicina y la dapsona en tres pacientes con recurrencia de lepra.

OBJECTIVE: To detect the presence of rifampin- and dapsone-resistant strains of Mycobacterium leprae in three patients with recurring leprosy and clinically-suspected antimicrobial resistance through molecular techniques. METHODS: A retrospective, descriptive study was conducted of three multibacillary patients at the "Agua de Dios" Sanitarium in Cundinamarca, Colombia, that presented leprosy relapses that were documented by medical history, bacilloscopy, and biopsy. Biopsies were taken of the skin lesions and the bacteria were subject to DNA extraction and purification. Regions of the rpoB and folP1 genes associated with antimicrobial resistance were amplified and subjected to touch-down polymerase chain reaction and the amplified products were sequenced using the Sanger method. RESULTS: A punctual mutation was identified in nucleotide 1367 of the rpoB gene in two of the samples studied. This mutation was not found in the folP1 gene of any of the three patients. CONCLUSIONS: The mutation identified showed strains of rifampin-resistant M. leprae in two of the three patients with recurring leprosy. Mutations that indicate dapsone-resistance were not detected in any of the three patients.

Estudio de resistencia a la rifampicina y la dapsona en tres pacientes con recurrencia de lepra / Study of rifampin and dapsone resistance in three patients with recurring leprosy

OBJETIVO: Detectar la presencia de cepas de Mycobacterium leprae resistentes a la rifampicina y la dapsona en tres pacientes con recurrencia de lepra y sospecha clínica de resistencia antimicrobiana, mediante la aplicación de técnicas moleculares. MÉTODOS: Se realizó un estudio descriptivo retrospectivo en tres pacientes multibacilares del Sanatorio de Agua de Dios, Cundinamarca, Colombia, que habían presentado recidivas de lepra documentadas por su historia clínica, baciloscopia y biopsia. Se obtuvieron biopsias de lesiones cutáneas que se procesaron para la extracción y purificación del ADN bacilar. Se amplificaron regiones de los genes rpoB y folP1 asociadas con la resistencia antimicrobiana, mediante la reacción en cadena de la polimerasa "touch-down" y se secuenciaron los productos amplificados mediante el método de Sanger. RESULTADOS: Se detectó una mutación puntual en el nucleótido 1367 del gen rpoB en dos de las muestras estudiadas. No se encontró la mutación estudiada en el gen folP1 en ninguno de los tres pacientes. CONCLUSIONES: La mutación identificada demostró la presencia de bacilos de M. leprae resistentes a la rifampicina en dos de los tres pacientes estudiados con recurrencia de la enfermedad. No se detectó la mutación indicadora de resistencia a la dapsona en ninguno de los tres pacientes.

OBJECTIVE: To detect the presence of rifampin- and dapsone-resistant strains of Mycobacterium leprae in three patients with recurring leprosy and clinically-suspected antimicrobial resistance through molecular techniques. METHODS: A retrospective, descriptive study was conducted of three multibacillary patients at the "Agua de Dios" Sanitarium in Cundinamarca, Colombia, that presented leprosy relapses that were documented by medical history, bacilloscopy, and biopsy. Biopsies were taken of the skin lesions and the bacteria were subject to DNA extraction and purification. Regions of the rpoB and folP1 genes associated with antimicrobial resistance were amplified and subjected to touch-down polymerase chain reaction and the amplified products were sequenced using the Sanger method. RESULTS: A punctual mutation was identified in nucleotide 1367 of the rpoB gene in two of the samples studied. This mutation was not found in the folP1 gene of any of the three patients. CONCLUSIONS: The mutation identified showed strains of rifampin-resistant M. leprae in two of the three patients with recurring leprosy. Mutations that indicate dapsone-resistance were not detected in any of the three patients.

[Guideline for the treatment of Hansen's disease in Japan (Second edition)].

ad hoc committee of Japanese Leprosy Association recommends revised standard treatment protocol of leprosy in Japan, which is a modification of World Health Organization's multidrug therapy (WHO/MDT, 1997). For paucibacillary (PB) leprosy, 6 months treatment by rifampicin and dapsone (MDT/PB) is enough. However, for high bacterial load multibacillary (MB) leprosy, 12 months treatment seems insufficient. Thus, (A) For MB with bacterial index (BI) > or = 3 before treatment, 2 years treatment by rifampicin, dapsone and clofazimine (MDT/MB) is necessary. When BI become negative and active lesion is lost within 2 years, no maintenance therapy is necessary. When BI is still positive, one year of MDT/MB is added (3 years in total), followed by maintenance therapy by dapsone and clofazimine until BI negativity and loss of active lesions. (B) For MB with BI < 3 or fresh MB (less than 6 months after the onset of the disease) with BI > or = 3, 1 year treatment by MDT/MB is necessary. When BI become negative and active lesion is lost within one year, no maintenance therapy is necessary. When BI is still positive or active lesion is remaining, additional therapy with MDT/MB for one more year is recommended. This is a simplification of first version in 2000. Brief summary of diagnosis, purpose of therapy, character of drugs, and prevention of deformity is also described.

[DNA microarray based rapid drug susceptibility test for Mycobacterium leprae].

Antibiotic susceptibility test of Mycobacterium leprae still relies on the time consuming methods based on the growth of M. leprae in the mouse footpad. Thus, the establishment of a rapid, simple and reliable method for the detection of drug-resistant M. leprae is one of the most urgent subjects in the treatment of leprosy patients. Recently, many data on the mutation of specific genes correlating with drug resistance have been accumulated. Application of these data permit the establishment of new gene diagnostic methods for drug susceptibility test of leprosy. In this paper, the method using the low density oligonucleotide array that enables the detection of base substitutions involved in resistance against anti-leprosy drugs on a single platform was discussed. The low density oligonucleotide array described in this paper will open the new perspectives in terms of patient management for leprosy with low cost requirement.