Evaluation of different molecular and phenotypic methods for identification of environmental Burkholderia cepacia complex

World J Microbiol Biotechnol. 2019 Feb 9;35(3):39. doi: 10.1007/s11274-019-2614-0.

Abstract

The correct identification of different genera and bacterial species is essential, especially when these bacteria cause infections and appropriate therapies need to be chosen. Bacteria belonging to the Burkholderia cepacia complex are considered important opportunistic pathogens, causing different types of infections in immunocompromised, principally in patients with cystic fibrosis. Twenty-one isolates were obtained from different soil samples and identified by sequencing of 16S rRNA, 23S rRNA, recA gene, MLST and by VITEK 2 and MALDI-TOF MS systems. Then, statistical analyses were performed. VITEK 2 and MALDI-TOF MS systems showed different bacterial genera. Sequencing of the 16S rRNA, 23S rRNA gene and amplification of recA gene showed that all the isolates belong to the B. cepacia complex. Sequencing of the recA gene showed a predominance of B. cenocepacia. The PCR of the recA gene showed a high specificity when it is necessary to identify the bacteria belonging to the B. cepacia complex in comparison with 16S and 23S rRNA genes sequencing. MLST analyzes showed a diversity of STs, which have not yet been correlated to the species. Phenotypic identification was not suitable for the identification of these pathogens since in many cases different genera have been reported, including identification by using MALDI-TOF MS.

Keywords: 16S rRNA; 23S rRNA; Burkholderia cepacia complex; MALDI-TOF MS; MLST; recA.

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Typing Techniques / methods*
  • Burkholderia Infections / microbiology
  • Burkholderia cepacia complex / classification*
  • Burkholderia cepacia complex / genetics*
  • Burkholderia cepacia complex / isolation & purification*
  • Cystic Fibrosis / microbiology
  • DNA, Bacterial / genetics
  • Multilocus Sequence Typing / methods
  • Opportunistic Infections / microbiology
  • Phenotype
  • Phylogeny*
  • Polymerase Chain Reaction / methods
  • RNA, Ribosomal, 16S / genetics
  • RNA, Ribosomal, 23S / genetics
  • Rec A Recombinases / genetics
  • Sequence Analysis, DNA
  • Soil Microbiology*
  • Species Specificity
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • RNA, Ribosomal, 16S
  • RNA, Ribosomal, 23S
  • Rec A Recombinases