Comparison between microsatellites and Ml MntH gene as targets to identify Mycobacterium leprae by PCR in leprosy

FUNDAMENTOS: PCR tem sido frequentemente utilizada no diagnóstico molecular da hanseníase. OBJETIVOS: comparar os resultados da PCR com 4 pares de primers específicos para Mycobacterium leprae, bem como os resultados da PCR à classificação operacional, segundo a OMS, de multibacilar (MB) e paucibacilar (PB) da hanseníase. MÉTODO: Vinte e oito amostras de DNA, extraído de biópsias congeladas de pele e de imprint de biópsias em papel de filtro de 23 pacientes (14 MB e 9 PB), foram utilizadas na PCR com primers que amplificam 131pb, 151pb e 168pb de regiões de microssatélites, e um fragmento de 336pb do gene Ml MntH (ML2098) do bacilo. RESULTADOS: O bacilo pôde ser detectado em 22 (78,6%) das 28 amostras. Nove (45%) das 20 amostras de biópsia e 6 (75%) das 8 amostras de imprints foram positivas para TTC. Sete (35,5%) amostras de biópsias e 5 (62,5%) imprints foram positivos para AGT, e 11 (55%) biópsias e 4 (50%) imprints foram positivos para AT. Oito (38%) amostras de biópsias e 5 (62,5%) imprints foram positivos para o gene Ml MntH. Dentre o grupo MB, os microssatélites detectaram o bacilo em 78,5% das amostras, e o gene Ml MntH, em 57,1% das amostras, independentemente do material clínico. No grupo PB, 55,5% das amostras foram positivas para os microssatélites, enquanto que 22,2% o foram para o gene Ml MntH. CONCLUSÕES: Estes resultados mostram que, tanto as regiões específicas de microssatélites quanto o gene Ml MntH, podem representar ferramentas úteis na detecção do Ml MntH por PCR em amostras de biópsias e imprint de biópsiasBACKGROUND: The Polymerase Chain Reaction (PCR) technique has been frequently used in the molecular diagnosis of leprosy. OBJECTIVES: To compare the results of PCR with four pairs of Mycobacterium leprae specific primers as well as to compare these results to multibacillary (MB) and paucibacillary (PB) leprosy according to the WHO operational classification. METHOD: 28 DNA samples, collected from the frozen skin biopsies and biopsy imprints on filter paper of 23 patients (14 MB and PB 9), were examined for PCR using primers which amplify 131, 151 and 168bp of specific microsatellite regions and a 336 fragment of the Ml MntH (ML2098) gene. RESULTS: M.leprae bacillus could be detected in 22 (78.6%) of the 28 samples. 9 (45%) of the 20 biopsy samples and 6 (75%) of the 8 imprints were positive to TTC. 7 (35.5%) skin biopsy specimens and 5 (62.5%) imprints were positive to AGT, and 11 (55%) biopsies and 4 (50%) were positive to AGT. 11 (55%) skin biopsies and 4 (50%) imprints were positive to AT. 8(38%) skin biopsies and 5 (62.5%) imprints were positive to the Ml MntH gene. In the MB group, the microsatellites detected the bacillus in 78.5% of the samples, and the Ml MntH gene in 57.1% of the samples, independent of the clinical material. In the PB group 55.5% of samples were positive to the microsatellite primers, while 22.2% were positive to the Ml MntH gene. CONCLUSIONS: These results show that both the specific regions of microsatellites, as well as the Ml MntH gene fragment can be useful tools for detecting the M. leprae DNA by PCR in frozen skin biopsy samples and filter paper biopsy imprintsCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)(FAEPA) Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto - Universidade de São Paulo (USP) - Fundação de Apoio ao Ensino, Pesquisa e Assistênci

Episódios reacionais da hanseníase podem ser exacerbados por infecções orais?

INTRODUCTION: This study evaluated whether leprosy reactions could be associated with oral infection. METHODS: Leprosy patients (n = 38) with (Group I) and without (Group II) oral infections were selected. Reactions were identified from the clinical and histopathological features associated with serum C-reactive protein (CRP) and10kDa interferon-gamma-induced protein (IP-10) levels, determined before and after elimination of the foci of infection. RESULTS: Group I presented more reactions than group II did, and improvement of the reactions after dental treatment. Serum CRP and IP-10 did not differ before and after the dental treatment, but differed between the groups. CONCLUSIONS: Oral infection could be an exacerbating factor in leprosy reactions.INTRODUÇÃO: Este estudo avaliou se as reações hansênicas podem estar associadas a infecções orais. MÉTODOS: Pacientes com hanseníase (n=38) com (Grupo I) e sem (Grupo II) infecções orais foram selecionados. As reações foram identificadas pelas características clínicas, histopatológicas, associadas a proteína-C-reativa (PCR) e proteína indutora de interferon-gamma de 10kDa (IP-10) séricos determinados antes e após a eliminação dos focos de infecção. RESULTADOS: Grupo I apresentou mais reações que o grupo II, e melhora das reações após o tratamento odontológico. PCR e IP-10 séricos não diferiram antes e após o tratamento odontológico, entretanto diferiram entre os grupos. CONCLUSÕES: As infecções orais podem ser exacerbadores das reações hansênicas.(CNPq) National Council for Scientific and Technological DevelopmentSão Paulo State Foundation against Leprosy(FAEPA) USP - Teaching, Research and Assistance Support Foundation of HCFMR

Leprosy-specific oral lesions : a report of three cases

Leprosy is a chronic infection caused by Mycobacterium leprae, a bacillus that presents a peculiar tropism for the skin and peripheral nerves. The clinical spectrum of leprosy ranges from the tuberculoid form (TT) to the disseminative and progressive lepromatous form (LL). Oral lesions are rare but, when present, occur in the lepromatous form. This article describes the clinical and microscopic findings of three cases of LL with oral manifestations. All patients had the lepromatous form and their leprosy-specific oral lesions occurred in the palate. The diagnosis was based on clinical, serological and histopathological findings, and multidrug therapy for multibacillary leprosy was started and continued for 24 months. All patients completed treatment, but developed reaction episodes which were treated with prednisone and/or thalidomide. The authors emphasize the importance of oral mucosa evaluation by a dental health professional during patient care since oral lesions may act as a source of infection

New sonographic measures of peripheral nerves: a tool for the diagnosis of peripheral nerve involvement in leprosy

To evaluate ultrasonographic (US) cross-sectional areas (CSAs) of peripheral nerves, indexes of the differences between CSAs at the same point (∆CSAs) and between tunnel (T) and pre-tunnel (PT) ulnar CSAs (∆TPTs) in leprosy patients (LPs) and healthy volunteers (HVs). Seventy-seven LPs and 49 HVs underwent bilateral US at PT and T ulnar points, as well as along the median (M) and common fibular (CF) nerves, to calculate the CSAs, ∆CSAs and ∆TPTs. The CSA values in HVs were lower than those in LPs (p 80%) and ∆TPT had the highest specificity (> 90%). New sonographic peripheral nerve measurements (∆CSAs and ∆TPT) provide an important methodological improvement in the detection of leprosy neuropathy