Discrimination of members of the Mycobacterium avium complex by polymerase chain reaction

Sircili, Marcelo Palma; Roxo, Eliana; Leão, Sylvia Cardoso

doi:10.1590/s0001-37141999000200011

Cited by 4 publications

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“…New algorithms were proposed by Taylor et al (27) and by Devalois et al (5), with the last including 34 species and subspecies. We have recently applied this technique to 18 MAC strains, including reference strains as well as animal and human isolates, and confirmed the usefulness of this technique in the differentiation of M. avium, M. intracellulare, and M. scrofulaceum (24).…”

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confidence: 68%

Identification of Two Novel Mycobacterium avium Allelic Variants in Pig and Human Isolates from Brazil by PCR-Restriction Enzyme Analysis

et al. 1999

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Mycobacterium avium complex (MAC) is composed of environmental mycobacteria found widely in soil, water, and aerosols that can cause disease in animals and humans, especially disseminated infections in AIDS patients. MAC consists of two closely related species, M. avium and M. intracellulare, and may also include other, less-defined groups. The precise differentiation of MAC species is a fundamental step in epidemiological studies and for the evaluation of possible reservoirs for MAC infection in humans and animals. In this study, which included 111 pig and 26 clinical MAC isolates, two novel allelic M. aviumPCR-restriction enzyme analysis (PRA) variants were identified, differing from the M. avium PRA prototype in theHaeIII digestion pattern. Mutations in HaeIII sites were confirmed by DNA sequencing. Identification of these isolates as M. avium was confirmed by PCR with DT1-DT6 and IS1245 primers, nucleic acid hybridization with the AccuProbe system, 16S ribosomal DNA sequencing, and biochemical tests. The characterization of M. avium PRA variants can be useful in the elucidation of factors involved in mycobacterial virulence and routes of infection and also has diagnostic significance, since they can be misidentified as M. simiae II and M. kansasii I if the PRA method is used in the clinical laboratory for identification of mycobacteria.

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Identification of Two Novel Mycobacterium avium Allelic Variants in Pig and Human Isolates from Brazil by PCR-Restriction Enzyme Analysis

et al. 1999

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“…1) (11,12,17,18) and are summarized in Table 1. Seventy-seven isolates (71.3%) were identified as M. avium PRA allelic variant I, 24 (22.2%) were identified as M. avium PRA allelic variant II, 6 (5.5%) were identified as M. avium PRA allelic variant III, and 1 (0.9%) was identified as M. avium PRA allelic variant IV.…”

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confidence: 99%

Identification of Mycobacterium avium Genotypes with Distinctive Traits by Combination of IS 1245 -Based Restriction Fragment Length Polymorphism and Restriction Analysis of hsp65

et al. 2003

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One-hundred eight Mycobacterium avium isolates from pigs, humans, birds, and bovines were typed by the IS1245-based restriction fragment length polymorphism (RFLP) method and PCR-restriction enzyme analysis (PRA) of hsp65. Nine clusters of isolates showing more than 80% similarity in their RFLP profiles were detected. The largest cluster (cluster B) included 32 of 79 pig isolates (40.5%), 3 of 25 human isolates (12%), and 1 of 2 bovine isolates, comprising 33% of all isolates. The second largest cluster (cluster A) included 18 pig isolates (22.8%) and 6 human isolates (24%). Six smaller clusters included six pig isolates (clusters C and D), four and two human isolates (clusters E and F, respectively), two pig isolates (cluster I), and two pig isolates plus one bovine isolate and the avian purified protein derivative strain (cluster H). Cluster G represented the "bird-type" profile and included the bird isolate in this series, one pig isolate, plus reference strain R13. PRA revealed four allelic variants. Seventy-seven isolates were identified as M. avium PRA variant I, 24 were identified as M. avium PRA variant II, 6 were identified as M. avium PRA variant III, and 1 was identified as M. avium PRA variant IV. Except for three isolates from cluster B, each of the RFLP clusters was associated with a single PRA pattern. Isolates with unique (nonclustered) RFLP profiles were distributed between PRA variants I and II, and there was one unique isolate of PRA variant IV. These observations are consistent with divergent evolution within M. avium, resulting in the emergence of distinct lineages with particular competence to infect animals and humans.

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“…O seqüenciamento direto do gene 16S de RNA ribossômico é um método rápido e acurado com grande poder discriminatório e amplamente utilizado para identificar e classificar diferentes espécies dentro do gênero Mycobacterium. Os genes que codificam a região 16S são estáveis e espécie-específicos, e os isolados também podem ser identificados e diferenciados pelas seqüências DT1 e DT6 por PCR (SIRCILI et al, 1998;COVERT et al, 1999;KOIVULA et al, 2004).…”

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Avaliação da virulência de isolados suínos de Mycobacterium avium no Brasil caracterizados pelo método de RFLP

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Ao Prof. Dr. José Soares Ferreira Neto, pela brilhante orientação, pela paciência e por me confiar a execução deste trabalho.À técnica Zenaide Maria de Morais, pelo auxílio durante a fase experimental, pelo apoio, amizade e dedicação constantes.Ao Prof. Dr. Fernando Ferreira e ao Prof. Dr. Ricardo Augusto Dias, pelo auxílio nas análises estatísticas deste trabalho.Ao funcionário Antonio da Costa Martins, pelos cuidados com os animais do biotério.À funcionária e sempre amiga Sandra Abelardo Sanches, pelo carinho, consideração, apoio e incentivo em todos os momentos de nossa convivência.Aos companheiros de pós-graduação da FMVZ e de outras unidades da USP, pela amizade e incentivo.A todos que, direta ou indiretamente, contribuíram para a conclusão deste trabalho.À CAPES, pela concessão temporária da bolsa de Doutorado. (respectively, p=0,15 and p=0,87). Inside the PIG A family, strain A52 was more virulent than strains A1 and A162; the strain C122 presented the lowest virulence compared with the strains C44 and C68 within the PIG C family. All strains, independent of their family, presented diverse virulence. Strain A52 was more virulent than strains A1, A162, B72, C122, D242 and D243. Distinct counting of CFU means different capacity of producing infection, i.e. diverse virulence. We concluded that, independent fo the genetic family established by similarity of RFLP patterns, M. avium isolates with diverse RFLP profiles may present different virulence; there was not difference in virulence; among all of four M. avium genetic families structured according to the similarity of RFLP patterns. RESUMO

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Discrimination of members of the Mycobacterium avium complex by polymerase chain reaction

Cited by 4 publications

References 10 publications

Identification of Two Novel Mycobacterium avium Allelic Variants in Pig and Human Isolates from Brazil by PCR-Restriction Enzyme Analysis

Identification of Two Novel Mycobacterium avium Allelic Variants in Pig and Human Isolates from Brazil by PCR-Restriction Enzyme Analysis

Identification of Mycobacterium avium Genotypes with Distinctive Traits by Combination of IS 1245 -Based Restriction Fragment Length Polymorphism and Restriction Analysis of hsp65

Avaliação da virulência de isolados suínos de Mycobacterium avium no Brasil caracterizados pelo método de RFLP

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