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Validation of the (GTG)5-rep-PCR fingerprinting technique for rapid classification and identification of acetic acid bacteria, with a focus on isolates from Ghanaian fermented cocoa beans

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Abstract
Amplification of repetitive bacterial DNA elements through the polymerase chain reaction (rep-PCR fingerprinting) using the (GTG)(5) primer, referred to as (GTG)(5)-PCR fingerprinting, was found a promising genotypic tool for rapid and reliable speciation of acetic acid bacteria (AAB). The method was evaluated with 64 AAB reference strains, including 31 type strains, and 132 isolates from Ghanaian, fermented cocoa beans, and was validated with DNA:DNA hybridization data. Most reference strains, except for example all Acetobacter indonesiensis strains and Gluconacetobacter liquefaciens LMG 1509, grouped according to their species designation, indicating the usefulness of this technique for identification to the species level. Moreover, exclusive patterns were obtained for most strains, suggesting that the technique can also be used for characterization below species level or typing of AAB strains. The (GTG)5-PCR fingerprinting allowed us to differentiate four major clusters among the fermented cocoa bean isolates, namely A. pasteurianus (cluster 1, 100 isolates), A. syzygii- or A. lovaniensis-like (cluster 11, 23 isolates), and A. tropicalis-like (clusters III and IV containing 4 and 5 isolates, respectively). A. syzygii-like and A. tropicalis-like strains from cocoa bean fermentations were reported for the first time. Validation of the method and indications for reclassifications of AAB species and existence of new Acetobacter species were obtained through 16S rRNA sequencing analyses and DNA:DNA hybridizations. Reclassifications refer to A. aceti LMG 153 1, Ga. xylinus LMG 1518, and Ga. xylinus subsp. sucrofermentans LMG 18788(T).

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Chicago
De Vuyst, Luc, N. Camu, T. De Winter, Katrien Vandemeulebroecke, V. Van de Perre, Marc Vancanneyt, Paul De Vos, and Ilse Cleenwerck. 2008. “Validation of the (GTG)5-rep-PCR Fingerprinting Technique for Rapid Classification and Identification of Acetic Acid Bacteria, with a Focus on Isolates from Ghanaian Fermented Cocoa Beans.” International Journal of Food Microbiology 125 (1): 79–90.
APA
De Vuyst, Luc, Camu, N., De Winter, T., Vandemeulebroecke, K., Van de Perre, V., Vancanneyt, M., De Vos, P., et al. (2008). Validation of the (GTG)5-rep-PCR fingerprinting technique for rapid classification and identification of acetic acid bacteria, with a focus on isolates from Ghanaian fermented cocoa beans. International Journal of Food Microbiology, 125(1), 79–90.
Vancouver
1.
De Vuyst L, Camu N, De Winter T, Vandemeulebroecke K, Van de Perre V, Vancanneyt M, et al. Validation of the (GTG)5-rep-PCR fingerprinting technique for rapid classification and identification of acetic acid bacteria, with a focus on isolates from Ghanaian fermented cocoa beans. International Journal of Food Microbiology. 2008;125(1):79–90.
MLA
De Vuyst, Luc, N. Camu, T. De Winter, et al. “Validation of the (GTG)5-rep-PCR Fingerprinting Technique for Rapid Classification and Identification of Acetic Acid Bacteria, with a Focus on Isolates from Ghanaian Fermented Cocoa Beans.” International Journal of Food Microbiology 125.1 (2008): 79–90. Print.
@article{626892,
  abstract     = {Amplification of repetitive bacterial DNA elements through the polymerase chain reaction (rep-PCR fingerprinting) using the (GTG)(5) primer, referred to as (GTG)(5)-PCR fingerprinting, was found a promising genotypic tool for rapid and reliable speciation of acetic acid bacteria (AAB). The method was evaluated with 64 AAB reference strains, including 31 type strains, and 132 isolates from Ghanaian, fermented cocoa beans, and was validated with DNA:DNA hybridization data. Most reference strains, except for example all Acetobacter indonesiensis strains and Gluconacetobacter liquefaciens LMG 1509, grouped according to their species designation, indicating the usefulness of this technique for identification to the species level. Moreover, exclusive patterns were obtained for most strains, suggesting that the technique can also be used for characterization below species level or typing of AAB strains. The (GTG)5-PCR fingerprinting allowed us to differentiate four major clusters among the fermented cocoa bean isolates, namely A. pasteurianus (cluster 1, 100 isolates), A. syzygii- or A. lovaniensis-like (cluster 11, 23 isolates), and A. tropicalis-like (clusters III and IV containing 4 and 5 isolates, respectively). A. syzygii-like and A. tropicalis-like strains from cocoa bean fermentations were reported for the first time. Validation of the method and indications for reclassifications of AAB species and existence of new Acetobacter species were obtained through 16S rRNA sequencing analyses and DNA:DNA hybridizations. Reclassifications refer to A. aceti LMG 153 1, Ga. xylinus LMG 1518, and Ga. xylinus subsp. sucrofermentans LMG 18788(T).},
  author       = {De Vuyst, Luc and Camu, N. and De Winter, T. and Vandemeulebroecke, Katrien and Van de Perre, V. and Vancanneyt, Marc and De Vos, Paul and Cleenwerck, Ilse},
  issn         = {0168-1605},
  journal      = {International Journal of Food Microbiology},
  language     = {eng},
  number       = {1},
  pages        = {79--90},
  title        = {Validation of the (GTG)5-rep-PCR fingerprinting technique for rapid classification and identification of acetic acid bacteria, with a focus on isolates from Ghanaian fermented cocoa beans},
  url          = {http://dx.doi.org/10.1016/j.ijfoodmicro.2007.02.030},
  volume       = {125},
  year         = {2008},
}

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