Article Options
Categories


Search


Advanced Search



This service is provided on D[e]nt Publishing standard Terms and Conditions. Please read our Privacy Policy. To enquire about a licence to reproduce material from endodonticsjournal.com and/or JofER, click here.
This website is published by D[e]nt Publishing Ltd, Phoenix AZ, US.
D[e]nt Publishing is part of the specialist publishing group Oral Science & Business Media Inc.

Creative Commons License


Recent Articles RSS:
Subscribe to recent articles RSS
or Subscribe to Email.

Blog RSS:
Subscribe to blog RSS
or Subscribe to Email.


Azerbaycan Saytlari

 »  Home  »  Endodontic Articles 1  »  A protocol for polymerase chain reaction detection of Enterococcus faecalis and Enterococcus faecium from the root canal
A protocol for polymerase chain reaction detection of Enterococcus faecalis and Enterococcus faecium from the root canal
Results



All strains of E. faecium and E. faecalis produced identical amplicon profiles with two major bands in positions corresponding to 320 and 420 base pairs (bp) (Fig. 1). When amplifying DNA of higher purity prepared from serial dilutions of E. faecalis using the Wizard Genomic DNA Purification System (Promega), a third band of 600 bp became evident (Fig. 2). The sequence of this previously undetected 600 bp amplicon was determined and aligned against the Genbank database. It matched no previously identified sequence but showed a partial (nucleotides 180–360 of 600) 90% homology to Xanthomonas campestris 16S-23S intergenic spacer DNA (Genbank acc. No. AF279434.1). S. equinus , S. uberis , S. milleri , S. anginosus , S. pyogenes , S. mutans , S. salivarius S. sanguis and G. morbillorum were associated with single bands in various positions (Figs 1, 3).
The DNA prepared directly from pure cultures of E. faecalis produced identical amplicon profiles as DNA prepared from frozen isolates.
The detection level of DNA in serial dilutions was 10 –13 grams (Fig. 4). The protocol for extraction of DNA resulted in a detection level of 10 cells (Fig. 2).


enterococcus, bacilli, bacterium, biological classification, enterococcus faecalis, medterms, medical dictionary, bacteria
Figure 1.PDR profiles of investigated strains. Lanes 1, E. faecalis (ATCC 19433, CCUG 19916); 2, E. faecium (ATCC 19434, CCUG 542); 3, E. faecalis (OMGS 350/ 98); 4, E. faecalis (OMGS 266/ 98); 5, E. faecalis (OMGS 349/98); 6, E. faecalis (OMGS 36798); 7, S. uberis; 8, S. equinus; 9, S. milleri; 10, S. mutans; 11, S. salivarius; 12, S. sanguis; 13, S. anginosus; 14, S. pyogenes; 15, G. morbillorum.
alcaligenes, enterococcus faecalis, alcaligenes faecalis, cephalosporins, burkholderiales, infectious substances, aminoglycosides
Figure 2.PCR profiles of 10-fold serial dilution series of cells of E. faecalis (ATCC 19433, CCUG 19916).
agarose, gel, dna, agar, band, fragment, agarose gel electrophoresis, electrophoresis, base pair, biochemistry, bromophenol blue, conformation, algae, anode, buffer solution, cresol red
Figure 3.PCR profiles of investigated strains and mixtures of strains run on a 2.5% TBE-agarose gel. A 20 bp interval ladder, ranging from 20 to 1000 bp, was used.
dilution series of extracted DNA
Figure 4.PCR profiles of 10-fold serial dilution series of extracted DNA from 107 cells of E. faecalis (ATCC 19433, CCUG 19916).