Folia Microbiologica

, Volume 56, Issue 1, pp 80–83 | Cite as

(GTG)5-PCR fingerprinting of lactobacilli isolated from cervix of healthy women

  • P. Švec
  • I. Sedláček
  • M. Chrápavá
  • P. Vandamme


A group of lactobacilli isolated from the cervix of 31 healthy women was characterized by (GTG)5-polymerase chain reaction (PCR) fingerprinting in order to evaluate this method for identification of vaginal lactobacilli. Obtained fingerprints were compared with profiles available in an in-house database of the CCM bacteria collection covering type and reference strains of multiple lactic acid bacteria including lactobacilli. Selected strains representing individual clusters were further identified by pheS gene sequencing. In total, six lactobacillus species were found among lactobacilli isolated from the cervix of healthy women. The (GTG)5-PCR method identified Lactobacillus gasseri (11 strains), Lactobacillus fermentum (one), and some of the Lactobacillus jensenii strains (eight out of 11), but failed to identify the remaining strains, including the Lactobacillus crispatus (18), Lactobacillus mucosae (one), and Lactobacillus vaginalis (one) species. L. jensenii strains were distributed over two fingerprint clusters. The majority of samples was dominated by one (GTG)5-PCR type. The rep-PCR fingerprinting using the (GTG)5 primer allowed straightforward identification of many, but not all, isolates. This method has been shown to be a useful tool for fast screening and grouping of vaginal lactobacilli, but its combination with another identification method is needed to obtain reliable identification results. In addition, Lactobacillus acidophilus was not shown to be the most common inhabitant of the female genital tract as generally assumed.


Lactobacillus Lactobacillus Acidophilus Lactobacillus Fermentum Fingerprint Profile Vaginal Lactobacillus 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



Financial support from Valosun Inc., state budget resources of the Ministry of Industry and Trade of the Czech Republic (project FI-IM5/205), and the Ministry of Education, Youth and Sports of the Czech Republic (project MSM0021622416) is acknowledged. We thank Ms. C. Snauwaert for technical support.


  1. Boyd MA, Antonio MAD, Hillier SL (2005) Comparison of API 50 CH strips to whole-chromosomal DNA probes for identification of Lactobacillus species. J Clin Microbiol 43:5309–5311PubMedCrossRefGoogle Scholar
  2. Brown CJ, Wong M, Davis CC, Kanti A, Zhou X, Forney LJ (2007) Preliminary characterization of the normal microbiota of the human vulva using cultivation-independent methods. J Med Microbiol 56:271–276PubMedCrossRefGoogle Scholar
  3. Dimitonova SP, Bakalov BV, Aleksandrova-Georgieva RN, Danova ST (2008) Phenotypic and molecular identification of lactobacilli isolated from vaginal secretions. J Microbiol Immunol Infect 41:469–477PubMedGoogle Scholar
  4. Euzéby JP (2010) List of prokaryotic names with standing in nomenclature. Accessed July 2010
  5. Gevers D, Huys G, Swings J (2001) Applicability of rep-PCR fingerprinting for identification of Lactobacillus species. FEMS Microbiol Lett 205:31–36PubMedCrossRefGoogle Scholar
  6. Kiss H, Kögler B, Petricevic L, Sauerzapf I, Klayraung S, Domig K, Viernstein H, Kneifel W (2007) Vaginal Lactobacillus microbiota of healthy women in the late first trimester of pregnancy. BJOG 114:1402–1407PubMedCrossRefGoogle Scholar
  7. Naser SM, Dawyndt P, Hoste B, Gevers D, Vandemeulebroecke K, Cleenwerck I, Vancanneyt M, Swings J (2007) Identification of lactobacilli by pheS and rpoA gene sequence analyses. Int J Syst Evol Microbiol 57:2777–2789PubMedCrossRefGoogle Scholar
  8. Pot B, Tsakalidou E (2009) Taxonomy and metabolism of Lactobacillus. In: Ljungh Å, Wadström T (eds) Lactobacillus molecular biology: from genomics to probiotics. Caister Academic Press, Norfolk, pp 3–58Google Scholar
  9. Pukall R (2006) DNA fingerprinting techniques applied to the identification, taxonomy and community analysis of prokaryotes. In: Stackebrandt E (ed) Molecular identification, systematics, and population structure of prokaryotes. Springer, Heidelberg, pp 51–82CrossRefGoogle Scholar
  10. Reid G (2009) Lactobacillus in the vagina: why, how, which ones, and what they do? In: Ljungh Å, Wadström T (eds) Lactobacillus molecular biology: from genomics to probiotics. Caister Academic Press, Norfolk, pp 183–196Google Scholar
  11. Scheirlinck I, Van der Meulen R, Van Schoor A, Vancanneyt M, De Vuyst L, Vandamme P, Huys G (2007) Influence of geographical origin and flour type on diversity of lactic acid bacteria in traditional Belgian sourdoughs. Appl Environ Microbiol 73:6262–6269PubMedCrossRefGoogle Scholar
  12. Švec P, Ševčíková A, Sedláček I, Bednářová J, Snauwaert C, Lefebvre K, Vandamme P, Vancanneyt M (2007) Identification of lactic acid bacteria isolated from human blood cultures. FEMS Immunol Med Microbiol 49:192–196PubMedCrossRefGoogle Scholar
  13. Švec P, Nováková D, Žáčková L, Kukletová M, Sedláček I (2008) Evaluation of (GTG)5-PCR for rapid identification of Streptococcus mutans. Antonie Leeuwenhoek 94:573–579PubMedCrossRefGoogle Scholar
  14. Švec P, Sedláček I, Žáčková L, Nováková D, Kukletová M (2009) Lactobacillus spp. associated with early childhood caries. Folia Microbiol 54:53–58CrossRefGoogle Scholar
  15. Van Hoorde K, Verstraete T, Vandamme P, Huys G (2008) Diversity of lactic acid bacteria in two Flemish artisan raw milk Gouda-type cheeses. Food Microbiol 25:929–935PubMedCrossRefGoogle Scholar
  16. Vásquez A, Jakobsson T, Ahrné S, Forsum U, Molin G (2002) Vaginal Lactobacillus flora of healthy Swedish women. J Clin Microbiol 40:2746–2749PubMedCrossRefGoogle Scholar

Copyright information

© Institute of Microbiology, v.v.i, Academy of Sciences of the Czech Republic 2011

Authors and Affiliations

  • P. Švec
    • 1
  • I. Sedláček
    • 1
  • M. Chrápavá
    • 2
  • P. Vandamme
    • 3
  1. 1.Czech Collection of Microorganisms, Department of Experimental Biology, Faculty of ScienceMasaryk UniversityBrnoCzech Republic
  2. 2.Valosun Inc.BrnoCzech Republic
  3. 3.Laboratory of MicrobiologyGhent UniversityGhentBelgium

Personalised recommendations