PCR primers for the amplification of the 16S rRNA gene of oral bacteria and for the specific identification of Actinobacillus actinomycetemcomitans.
The aim of this study was to develop a PCR reaction specific to Actinobacillus actinomycetemcomitans, which targets a widely conserved gene of this bacterium. Two sets of primers were designed based on published sequences of the 16S rRNA of several microorganisms. The first set amplifies a major part of the 16S small subunit rRNA gene of several strains of bacteria commonly found in the periodontal pocket. This reaction produced a 1306 bp-long product and served as a positive control. The second set was specific to A. actinomycetemcomitans and produced a 449 bp-long product. H. aphrophilus and E. coli yielded positive results with the control primers and negative results with the A. actinomycetemcomitans-specific primers. DNA-DNA hybridization was used to validate the identify of the amplified sequences. B. cereus, which is a common contaminator in the laboratory, and human DNA did not generate PCR products in either reaction. The developed primers seen useful for the identification of A. actinomycetemcomitans strains.
Albandar
J M
JM
Department of Periodontology, Dental Faculty, University of Oslo, Norway. albandarj@de45.nidr.nih.gov
Lyngstadaas
S P
SP
Forbord
B
B
D016976:Aggregatibacter actinomycetemcomitans / Q000235:genetics* / Q000302:isolation & purification*; D015373:Bacterial Typing Techniques; D001483:Base Sequence; D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics*; D005798:Genes, Bacterial; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S