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1.
PLoS Negl Trop Dis ; 2(11): e328, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18982056

RESUMO

Mycobacterium leprae is not cultivable in axenic media, and direct microscopic enumeration of the bacilli is complex, labor intensive, and suffers from limited sensitivity and specificity. We have developed a real-time PCR assay for quantifying M. leprae DNA in biological samples. Primers were identified to amplify a shared region of the multicopy repeat sequence (RLEP) specific to M. leprae and tested for sensitivity and specificity in the TaqMan format. The assay was specific for M. leprae and able to detect 10 fg of purified M. leprae DNA, or approximately 300 bacteria in infected tissues. We used the RLEP TaqMan PCR to assess the short and long-term growth results of M. leprae in foot pad tissues obtained from conventional mice, a gene knock-out mouse strain, athymic nude mice, as well as from reticuloendothelial tissues of M. leprae-infected nine-banded armadillos. We found excellent correlative results between estimates from RLEP TaqMan PCR and direct microscopic counting (combined r = 0.98). The RLEP TaqMan PCR permitted rapid analysis of batch samples with high reproducibility and is especially valuable for detection of low numbers of bacilli. Molecular enumeration is a rapid, objective and highly reproducible means to estimate the numbers of M. leprae in tissues, and application of the technique can facilitate work with this agent in many laboratories.


Assuntos
Mycobacterium leprae/genética , Animais , Calibragem , DNA Bacteriano/genética , DNA Ribossômico/genética , Escherichia coli/genética , Membro Posterior/microbiologia , Camundongos , Camundongos Nus/microbiologia , Mycobacterium leprae/crescimento & desenvolvimento , Mycobacterium leprae/isolamento & purificação , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética , Staphylococcus epidermidis/genética , Streptococcus pyogenes/genética
3.
Kitasato Arch Exp Med ; 64(4): 213-20, 1991 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-1823921

RESUMO

Mycobacterium leprae is an obligate intracellular parasite which grows within mononuclear phagocytes. In clinical cases, M. leprae reaches enormous numbers in the macrophage-rich granulomas of leprosy. Peritoneal macrophages from CBA mice were cultured in Waymouth medium containing fresh horse serum in Costar 3424 trays (24 wells, 16 mm in diameter) each containing 9 x 12 mm cover slips. This medium was supplemented with 0.5 micrograms/ml of cycloheximide. These cells were infected with M. leprae Thai-53 strain obtained by nude mice inoculation. Significant multiplication of the acid-fast bacilli in the macrophages was observed three weeks after inoculation. This experiment showed M. leprae mainly multiplied in cells and not by rephagocytization of M. leprae derived from destroyed cells.


Assuntos
Técnicas Bacteriológicas , Macrófagos/microbiologia , Mycobacterium leprae/crescimento & desenvolvimento , Animais , Técnicas Bacteriológicas/instrumentação , Divisão Celular/efeitos dos fármacos , Cicloeximida/farmacologia , Feminino , Macrófagos/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos CBA/microbiologia , Camundongos Nus/microbiologia , Fagocitose
4.
Int J Lepr Other Mycobact Dis ; 55(4 Suppl): 889-95, 1987 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-3325583

RESUMO

Two aspects of the immune deficiency of nude mice make these animals particularly useful tools for leprosy research. Nude mice are capable of supporting multiplication of M. leprae to levels approaching 10(10) per g in peripheral body tissues. In addition, nude mice may be inoculated with greater than 10(4) (in fact, with as many as 10(8) organisms per foot pad, without provoking an immune response that prevents multiplication of the organisms. Thus, the nude mouse should be particularly suitable for detecting persisting M. leprae in treated patients, and as a model of the patient for evaluating chemotherapeutic regimens.


Assuntos
Hanseníase/imunologia , Camundongos Nus/microbiologia , Mycobacterium leprae/crescimento & desenvolvimento , Animais , Hanseníase/tratamento farmacológico , Camundongos , Rifampina/uso terapêutico
7.
s.l; s.n; 1984. 4 p. tab, graf.
Não convencional em Inglês | SES-SP, SES-SP, HANSEN, HANSENIASE, SESSP-ILSLACERVO, SES-SP | ID: biblio-1240388

RESUMO

The continued failure to grow the leprosy bacillus in vitro and the limited and localized nature of the infection resulting from the inoculation of Mycobacterium leprae into normal mice emphasise the need for an experimental animal which readily permits growth of the bacillus.


Assuntos
Camundongos , Animais , Camundongos Nus/microbiologia , Hanseníase/fisiopatologia , Modelos Animais de Doenças
10.
Int J Lepr Other Mycobact Dis ; 50(1): 68-75, 1982 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-7042613

RESUMO

Ultrastructural features of the growth of M. leprae and M. lepraemurium in nude mouse macrophages were studied by ultrathin sectioning and freeze-etching. In nude mouse macrophages, M. leprae produced spherical droplets (foamy structures) similar to those in human lepra cells. On the other hand, M. lepraemurium produced typical crystalline material in nude mouse macrophages, which is quite the same as that observed in the C3H strain mouse, Spherical droplets in the form of foamy structures seem to be made up of a specific substance produced by the multiplication of M. leprae in suitable host cells (human, nude mouse, and armadillo macrophages).


Assuntos
Mycobacterium leprae/ultraestrutura , Mycobacterium lepraemurium/ultraestrutura , Animais , Humanos , Macrófagos/microbiologia , Camundongos , Camundongos Endogâmicos C3H , Camundongos Nus/microbiologia , Microscopia Eletrônica , Mycobacterium leprae/crescimento & desenvolvimento , Mycobacterium lepraemurium/crescimento & desenvolvimento , Pele/ultraestrutura
11.
Int J Lepr Other Mycobact Dis ; 50(1): 83-9, 1982 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-7042615

RESUMO

Various procedures to decontaminate and purify M leprae free of host tissue material resulted in total retention of their intracellular ATP and also infectiousness. The ATP content of one million M. leprae cells, isolated from either livers, spleens, or lymph nodes of infected armadillos, or a nude mouse foot pad or a human biopsy specimen, was in the range of 1.17 to 1.40 picograms. Suspensions could be decontaminated with 4% NaOH and all non-bacterial ATP could be eliminated by the combined action of trypsin, chymotrypsin, and collagenase initially, followed by Triton X-100 plus ATPase. These findings further assure that M. leprae are different from M. lepraemurium in that they can withstand even the severest purification procedures that are necessary in order for them to be used for sophisticated biochemical and metabolic studies.


Assuntos
Trifosfato de Adenosina/análise , Mycobacterium leprae/análise , Trifosfato de Adenosina/isolamento & purificação , Animais , Tatus/microbiologia , Humanos , Camundongos , Camundongos Nus/microbiologia , Mycobacterium lepraemurium/análise
13.
Jpn J Exp Med ; 46(3): 167-80, 1976 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-785061

RESUMO

Comparative observations were made on the development of experimental murine leprosy in various inbred strains of mice, including nude mice of congenital thymic aplasia. The susceptibility of these strains of mice to murine leprosy bacilli was evaluated by the development of leproma at the subcutaneous infection site and also by the involvement of visceral organs. Nude mice developed a much more severe disease than C3H mice which is the representative of the malignant type. Their high sensitivity was also demonstrated in the case of intraperitoneal infection. The observations in nude mice and other mouse strains confirmed our concept that experimental mouse leprosy can be classified into three clinical types, benign, intermediate and malignant, and suggested that such mouse strain differences are related with their cell-mediated immunity.


Assuntos
Infecções por Mycobacterium/imunologia , Mycobacterium lepraemurium/imunologia , Animais , Granuloma/patologia , Imunidade Celular , Fígado/patologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Camundongos Nus/microbiologia , Infecções por Mycobacterium/patologia , Pele/patologia , Neoplasias Cutâneas/patologia , Úlcera Cutânea/patologia , Baço/patologia
14.
In. Annual Leprosy Research Conference, 4. Annual Leprosy Research Conference, 4/Abstracts of papers. San Francisco, U.S. Public Health Service Hospital, 1969. p.458.
Não convencional em Inglês | SES-SP, SES-SP, HANSEN, HANSENIASE, SESSP-ILSLACERVO, SES-SP | ID: biblio-1243387
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