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TitleIn silico vs in vitro analysis of primer specificity for the detection of Gardnerella vaginalis, atopobium vaginae and lactobacillus spp
Author(s)Henriques, Ana Filipa Frutuoso Mendes
Cereija, Tatiana Barros Reis
Machado, António
Cerca, Nuno
KeywordsA. vaginae
G. vaginalis
In silico primer testing
In vitro primer testing
Lactobacillus spp.
Primer sensitivity
Primer specificity
Issue date2012
PublisherBioMed Central (BMC)
JournalBMC Research Notes
Abstract(s)Bacterial vaginosis (BV) is a common pathology of women in reproductive age that can lead to serious health complications, and is associated with shifts in the normal microflora from predominance of Lactobacillus spp. to a proliferation of other anaerobes such as G. vaginalis and A vaginae, which can be detected by PCR. The optimal PCR pathogen detection assay relies mainly on the specificity and sensitivity of the primers used. Findings Here we demonstrate that in silico analytical testing of primer specificity is not a synonym to in vitro analytical specificity by testing a range of published and newly designed primers with both techniques for the detection of BV-associated microorganisms. Conclusions By testing primer in vitro specificity with a sufficient range of bacterial strains, we were able to design primers with higher specificity and sensitivity. Also by comparing the results obtained for the newly designed primers with other previously published primers, we confirmed that in silico analysis is not sufficient to predict in vitro specificity. As such care must be taken when choosing the primers for a detection assay.
AccessOpen access
Appears in Collections:CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series

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