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Bacteriology

Discrimination of Burkholderia multivorans and Burkholderia vietnamiensis fromBurkholderia cepacia Genomovars I, III, and IV by PCR

Adolf Bauernfeind, Ines Schneider, Renate Jungwirth, Carsten Roller
Adolf Bauernfeind
Max von Pettenkofer-Institut, Munich, Germany
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Ines Schneider
Max von Pettenkofer-Institut, Munich, Germany
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Renate Jungwirth
Max von Pettenkofer-Institut, Munich, Germany
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Carsten Roller
Max von Pettenkofer-Institut, Munich, Germany
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DOI: 
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Tables

  • Table 1.

    Investigated strains

    OrganismStrain designation or origina
    B. vietnamiensis LMG 10929T, LMG 6998, LMG 6999, CF isolateb
    B. cepacia DSM 50181, ATCC 25416T, LMG 7000, LMG 12614, LMG 12615, LMG 14294, CF isolatesc
    B. multivorans LMG 14280, LMG 14293, CF isolatesd
    B. gladioli DSM 4285, CF isolatese
    B. mallei ATCC 23344T, ATCC 15310, ATCC 10399
    B. pseudomallei ATCC 23343T, ATCC 15682, NCTC 1691, clinical isolatesf
    R. pickettii ATCC 27511T, clinical isolateg
    • ↵a DSM, Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Braunschweig, Federal Republic of Germany; LMG, Laboratorium voor Microbiologie, Universiteit Gent, Gent, Belgium; ATCC, American Type Culture Collection, Rockville, Md.; NCTC, National Collection of Type Cultures, Central Public Health Laboratory, London, England.

    • ↵b Strain from The Netherlands (n = 1).

    • ↵c Strains from Germany (n = 34), The Netherlands (n = 2), and the United Kingdom (n = 6).

    • ↵d Strains from Germany (n = 60), The Netherlands (n = 11), and Denmark (n = 8).

    • ↵e Strains from Germany (n = 2) and Canada (n = 3).

    • ↵f Strains from Germany (n = 1) and Switzerland (n = 1).

    • ↵g Strain from Germany (n = 1).

  • Table 2.

    Primers selected for PCR

    Target speciesPrimer designationaSequencebAnnealing temp (°C)
    Eubacteria Eub-16-15′-AGR GTT YGA TYM TGG CTC AG-3′56
    Eubacteria Eub-16-25′-ACC GCG GCT GCT GGC AC-3′60
    Burkholderia spp. and Ralstoniaspp.BuRa-16-15′-AGC ACT TTT GTC CGG RAA-3′52
    Burkholderia spp. and Ralstoniaspp.BuRa-16-25′-AAT CCC CAA CAA CTA GTT G-3′54
    B. cepacia, B. multivorans, and B. vietnamiensis CeMuVi-16-2457 5′-CCG RCT GTA TTA GAG CCA-3′55
    B. gladioli Gl-16-2457 5′-CGA AGG ATA TTA GCC CTC-3′54
    B. cepacia Ce-16-21028 5′-AGC ACT CCC RCC TCT CAG-3′c 59
    B. multivorans andB. vietnamiensis MuVi-16-21028 5′-AGC ACT CCC GAA TCT CTT-3′54
    B. mallei and B. pseudomallei MaPs-16-2457 5′-CTC CGG GTA TTA GCC AGA-3′56
    Ralstonia pickettii Pi-16-2461 5′-TCG ACC CCA GGT ATT AAC-3′54
    B. vietnamiensis, B. mallei, and B. pseudomallei ViMaPs-23-15′-CCT TTG GGT CAT CCT GGA-3′56
    B. cepacia and B. vietnamiensis CeVi-23-25′-TCC TAC CAT GCG TGC AA-3′52
    B. mallei Ma-23-25′-CAC CGA AAC TAG CA-3′42
    • ↵a 16, 16S rDNA; 23, 23S rDNA; 1, sense (forward) primer; and 2, antisense (backward) primer.

    • ↵b Wobble bases: M = A or C; R = A or G; and Y = C or T.

    • ↵c Partially congruent with primer PSR1 of Campbell et al. (11), i.e., 5′-TTTCGAGCACTCCCGCCTCTCAG-3′.

  • Table 3.

    Specificity of primer combinations for the identification of Burkholderia spp.

    Primer designationaTarget organismsSize of PCR product (bp) for:
    B. cepaciagenomovarB. multivoransB. vietnamiensisB. gladioliB. malleiB. pseudomalleiR. pickettii
    IIIIIV
    Eub-16-1 + Eub-16-2 Eubacteria 519519519519519519519519519
    BuRa-16-1 + BuRa-16-2 Burkholderia andRalstoniaspp.409409409409409409409409409
    Eub-16-1 + CeMuVi-16-2457 B. cepacia,B. multivorans, and B. vietnamiensis 463463463463463
    Eub-16-1 + Ce-16-21028 B. cepacia 1,0151,0151,015
    Eub-16-1 + MuVi-16-21028 B. multivorans and B. vietnamiensis 1,0151,015
    ViMaPs-23-1 + CeVi-23-2 B. vietnamiensis 1,055
    Eub-16-1 + Gl-16-2457 B. gladioli 463
    Eub-16-1 + MaPs-16-2457 B. mallei and B. pseudomallei 463463
    ViMaPs-23-1 + Ma-23-2 B. mallei 526
    Eub-16-1 + Pi-16-2461 R. pickettii 467
    • ↵a For sequences see Table 2.

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Discrimination of Burkholderia multivorans and Burkholderia vietnamiensis fromBurkholderia cepacia Genomovars I, III, and IV by PCR
Adolf Bauernfeind, Ines Schneider, Renate Jungwirth, Carsten Roller
Journal of Clinical Microbiology May 1999, 37 (5) 1335-1339; DOI:

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Discrimination of Burkholderia multivorans and Burkholderia vietnamiensis fromBurkholderia cepacia Genomovars I, III, and IV by PCR
Adolf Bauernfeind, Ines Schneider, Renate Jungwirth, Carsten Roller
Journal of Clinical Microbiology May 1999, 37 (5) 1335-1339; DOI:
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