Article Figures & Data
Figures
- FIG. 1.
Phylogenetic tree comparing sequences of a 429-bp fur fragment from strains and isolates of the BCC. The phylogenies are rooted due to the assumption of a common ancestor and biological clock. Genetic distance is shown on the scale, with demarcations representing 2% estimated substitutions. The genomovar status (I through IX) of each BCC species is shown in parentheses following the strain or isolate name. Abbreviations are as follows: Bg, Burkholderia gladioli; Bp, Burkholderia pseudomallei; Bsp, Burkholderia species; Bx, Burkholderia xenovorans.
- FIG. 2.
Gel electrophoresis of fur PCR products for the BCC species B. vietnamiensis, B. dolosa, B. ambifaria, B. anthina, and B. pyrrocinia. PCR products were electrophoresed on a 0.8% agarose gel prior to visualization with ethidium bromide. Molecular weight standards (MW) are shown in lane 1, with the corresponding base pair sizes shown at the left of the gel. Shown below each BCC species name is the fur primer set used and the size of the obtained PCR product (in base pairs) with the following template DNAs: lane 2, PC259/LMG 18835; lane 3, G4; lane 4, ATCC 29424; lane 5, STM1441/LMG 21443; lane 6, CEP021/LMG 21819; lane 7, E12/LMG 21820; lane 8, L06; lane 9, CEP0996/LMG 19467; lane 10, M53; lane 11, J2552/LMG 16670; lane 12, C1765/LMG 20983; lane 13, AU1293/LMG 21821; lane 14, ATCC 39277/LMG 21822; lane 15, BC011/LMG 21823; lane 16, C1469/LMG 21824.
Tables
- TABLE 1.
Strains and clinical isolates used in this studya
Species Strain/isolate Source Location recA RFLP result Growth on BCSA Amplification with JD490/JD491 Amplification with species-specific primers Burkholderia cepacia CEP509*/LMG 18821* CF Australia E NT + + ATCC 25416T*/LMG 1222T* ENV United States D NT + + ATCC 17759*/LMG 2161* ENV Trinidad E NT + + BCC group K CEP0964 CF Canada K NT + + CEP1056 CF Canada K NT + + R445 CF Canada + + + Burkholderia multivorans LMG 13010 T* CF Belgium NT + + ATCC 17616*/LMG 17588* ENV United States F NT + + C3430 CF Canada NT + + C5274 CF Canada NT + + C5393*/LMG 18822* CF Canada F NT + + C5568 CF Canada NT + + M1512 CF Canada + + + M1865 CF Canada + + + R810 CF Canada + + + R1159 CF Canada + + + Burkholderia cenocepacia K56-2*/LMG 18863* CF-e Canada G NT + + J2315*/LMG 16656* CF-e United Kingdom G NT + + C1257 CF-e Canada NT + + C4455 CF-e Canada NT + + C5424*/LMG 18827* CF-e Canada G NT + + C6433*/LMG 18828* CF-e Canada NT + + CEP511*/LMG 18830* CF-e Australia I NT + + D1 ENV United States NT + + PC184*/LMG 18829* CF-e United States J′ NT + + LMG 19240 ENV Australia NT + − CEP0868/LMG 21461 CF Argentina NT + + R161 CF Canada + + + R452 CF Canada + + + R750 CF Canada + + + R1284 CF Canada + + + R1285 CF Canada + + + R1314 CF Canada + + + R1434 CF Canada + + + R1619 CF Canada + + + R1882 CF Canada + + + R1883 CF Canada + + + R1884 CF Canada + + + R2314 CF Canada + + + S11528 CF Canada + + + Burkholderia stabilis LMG 14294 T* CF Belgium J NT + + C7322*/LMG 18870* CF Canada NT + + R450 CF Canada + + + R2140 CF Canada + + + R2339 CF Canada + + + Burkholderia vietnamiensis LMG 10929 T* ENV Vietnam B NT + + PC259*/LMG 18835* CF United States A NT + + ATCC 29424 ENV United States NT + + G4 ENV United States A NT + + Burkholderia dolosa AU0645 T*/LMG 18943T* CF United States Q NT + + CEP021*/LMG 21819* CF United States NT + + E12*/LMG 21820* CF United Kingdom NT + + L06 CF NT + + STM1441*/LMG 21443* ENV Senegal NT + + Burkholderia ambifaria ATCC 53266*/LMG 17828* ENV United States L NT + + AMMDT*/LMG 19182T* ENV United States N NT + + CEP0996*/LMG 19467* CF Australia N NT + + M53 ENV United States NT + + Burkholderia anthina W92 T*/LMG 20980T* ENV United States T NT + + C1765*/LMG 20983* CF United Kingdom T NT + + J2552*/LMG 16670* ENV United Kingdom AS NT + + AU1293*/LMG 21821* CF United States AS NT + + Burkholderia pyrrocinia ATCC 15958 T*/LMG 14191T* ENV Japan NT + + ATCC 39277*/LMG 21822* ENV United States P NT + + BC011*/LMG 21823* ENV United States AR NT + + C1469*/LMG 21824* CF United Kingdom AA NT + + Burkholderia gladioli R406 CF Canada NA + + NA R1879 CF Canada NA + + NA Pseudomonas aeruginosa R285 CF Canada NA − − NA Herbaspirillum sp. R740 CF Canada NA − − NA Listeria monocytogenes R1653 CF Canada NA − − NA Pandoraea sp. R1717 CF Canada NA + − NA Burkholderia sp. JS150 ENV United States NA NT + NA ↵ a Abbreviations: RFLP, restriction fragment length polymorphism; CF, CF isolate; ENV, environmental isolate; CF-e, CF epidemic isolate; NA, not applicable; NT, not tested. Strains in boldface type were used to test each of the nine species-specific primer sets. Asterisks indicate strains of the BCC experimental strain panel (10, 22).
- TABLE 2.
Species-specific primers used for amplification of the BCC fur gene
Species Forward primer Forward primer sequence Forward primer positions Reverse primer Reverse primer sequence Reverse primer positions Product length (bp) BCC species amplified B. cepacia/BCC group K F1 GGCNGAAGACGTCTACCGG 102-120 R1 TCGAAGTTGCTGCGCGAC 201-218 117 B. cepacia B. multivorans F2 AGCAGAGCCCCGTGCGG 77-93 R2 GGTGGGGGCAGTTTTCGGTG 399-418 342 B. multivorans B. cenocepacia F TGACCAATCCGACCGATCTCA 2-22 R3 ATCGCCTGCTGGCGGCTC 321-338 337 B. cenocepacia IIIA, IIIB, IIID B. stabilis F4 CNACCGTCTATCGCGTGCTC 155-174 R TCAGTGCTTGCGGTGGGG 412-429 275 B. multivorans, B. cenocepacia, B. stabilis, B. dolosa B. vietnamiensis F TGACCAATCCGACCGATCTCA 2-22 R5 CGTGGTGGGAGCCTTCGTTG 243-262 261 B. vietnamiensis B. dolosa F6 CTAAAGGCCACCCTACCGCGG 34-54 R TCAGTGCTTGCGGTGGGG 412-429 396 B. dolosa B. ambifaria F7 CCCNGTGCGTCACCTGACT 84-102 R7 CGTGGTGCGAACCTTCATTCAA 241-262 179 B. ambifaria B. anthina F TGACCAATCCGACCGATCTCA 2-22 R8 CAGGTGACGCACGGGGCTC 81-99 98 B. multivorans, B. cenocepacia, B. stabilis, B. anthina, B. pyrrocinia (1 strain) B. pyrrocinia F1 GGCNGAAGACGTCTACCGG 102-120 R9 ATCGCCTGCTGGCGGCC 322-338 237 B. pyrrocinia JD490 ATGACCAATCCGACCGATCTCAA 1-23 JD491 TCAGTGCTTGCGITNIGGGCAGTT 406-429 429 All
