Fluorescence in situ Hybridization method using Peptide Nucleic Acid probes for rapid detection of Lactobacillus and Gardnerella spp.

BMC Microbiology

Research Article

Ligia Rodrigues¹  Maria João Vieira¹  Nuno Cerca¹  Débora Salgueiro¹  António Machado¹  Ana Henriques¹  Carina Almeida²  Nuno Filipe Azevedo³  Mario Vaneechoutte⁴  Freddy Haesebrouck⁵ 

[1] IBB - Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal;IBB - Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, University of Minho, Campus de Gualtar, 4710-057, Braga, Portugal;LEPAE, Department of Chemical Engineering, Faculty of Engineering, University of Porto, rua Dr. Roberto Frias, 4200-465, Porto, Portugal;LEPAE, Department of Chemical Engineering, Faculty of Engineering, University of Porto, rua Dr. Roberto Frias, 4200-465, Porto, Portugal;Laboratory of Bacteriology Research, Faculty Medicine & Health Sciences, University of Ghent, B-9000, Ghent, Belgium;Laboratory of Veterinary Bacteriology and Mycology, Faculty of Veterinary Medicine, Ghent University, B9820, Merelbeke, Belgium;

关键词: in situ;    Peptide Nucleic Acid Probe (PNA probe);    Lactobacillus;    Gardnerella vaginalis;    Bacterial vaginosis;   

DOI  :  10.1186/1471-2180-13-82

 received in 2012-11-26, accepted in 2013-03-27,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundBacterial vaginosis (BV) is a common vaginal infection occurring in women of reproductive age. It is widely accepted that the microbial switch from normal microflora to BV is characterized by a decrease in vaginal colonization by Lactobacillus species together with an increase of Gardnerella vaginalis and other anaerobes. Our goal was to develop and optimize a novel Peptide Nucleic Acid (PNA) Fluorescence in situ Hybridization assay (PNA FISH) for the detection of Lactobacillus spp. and G. vaginalis in mixed samples.ResultsTherefore, we evaluated and validated two specific PNA probes by using 36 representative Lactobacillus strains, 22 representative G. vaginalis strains and 27 other taxonomically related or pathogenic bacterial strains commonly found in vaginal samples. The probes were also tested at different concentrations of G. vaginalis and Lactobacillus species in vitro, in the presence of a HeLa cell line. Specificity and sensitivity of the PNA probes were found to be 98.0% (95% confidence interval (CI), from 87.8 to 99.9%) and 100% (95% CI, from 88.0 to 100.0%), for Lactobacillus spp.; and 100% (95% CI, from 92.8 to 100%) and 100% (95% CI, from 81.5 to 100.0%) for G. vaginalis. Moreover, the probes were evaluated in mixed samples mimicking women with BV or normal vaginal microflora, demonstrating efficiency and applicability of our PNA FISH.ConclusionsThis quick method accurately detects Lactobacillus spp. and G. vaginalis species in mixed samples, thus enabling efficient evaluation of the two bacterial groups, most frequently encountered in the vagina.

【 授权许可】

Unknown   
© Machado et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.