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Deoxyribonucleic acid relatedness between Mycobacterium leprae or Mycobacterium lepraemurium and other selected bacteria was studied by both dot blot and spectrophotometric deoxyribonucleic acid hybridization assays. The results obtained by the two methods were similar, except for the relatedness values between M. leprae and two corynebacterial strains. Among the mycobacterial species examined, acid-fast organisms isolated from armadillos and a mangabey monkey with leprosy-like disease showed 100% relatedness with M. leprae grown experimentally in armadillos, suggesting their common origin. In this study we demonstrate the usefulness of the dot blot hybridization technique as a screening method for mycobacterial taxonomy. The deoxyribonucleic acids (DNAs) of Mycobacterium leprae and Mycobacterium lepraemurium were analyzed kinetically to determine their genome sizes, base ratios, and genetic relatedness to certain corynebacteria, mycobacteria, and nocardiae (5, 6). M. leprae grown in armadillos was found to be a unique organism among the Corynebacterium- Mycobacterium-Nocardia group, whereas M. lepraemurium grown in mice showed close genetic relatedness to Mycobac- terium avium. These studies demonstrated the usefulness of DNA relatedness values in taxonomic or phylogenetic stud- ies of the corynebacteria, mycobacteria, and nocardiae. However, one of the disadvantages of the kinetic analysis used in these studies was the requirement for extremely purified DNA samples in amounts of at least 1 mg to determine the DNA relatedness between the test species and 20 other organisms. On the other hand, the dot blot hybrid- ization method (7) requires neither extreme purity nor large amounts of DNA samples. Since the supply of M. leprae is seriously limited, this latter method facilitates the study of the DNA relatedness of this organism. In this study we determined the DNA relatedness between M. leprae or M. lepraemurium and other selected organisms belonging to the Corynebacterium-Mycobacterium-Nocardia group by using dot blot hybridization as a screening test in comparison with a spectrophotometric assay. Bacterial strains and culture conditions. The bacterial strains used in this study and their sources are listed in Tables 1 and 2. The M. leprae strain which was used as the standard strain in the kinetic analysis and also as a source of the labeled probe in dot blot hybridization experiments was isolated from cutaneous nodules of armadillo A24 infected with M. leprae of human origin. This tissue was prepared by E. E. Storr, Medical Research Institute of Florida Institute of Technology, and was supplied through the National Institute of Allergy and Infectious Diseases. An acid-fast organism, which is listed as M. leprae (indigenous) in Table 1, was isolated from spleen tissues of two wild armadillos (armadillos V11 and V69) with leprosy-like disease. These infected tissues were supplied by W. F. Kirchheimer, Na-

international journal of systematic bacteriology

Deoxyribonucleic Acid Relatedness Among Mycobacterium leprae, Mycobacterium lepraemurium, and Selected Bacteria by Dot Blot and Spectrophotometric Deoxyribonucleic Acid Hybridization Assays.