Identification of Actinobacillus actinomycetemcomitans using species-specific 16S rDNA primers.
The purpose of this study was to develop species-specific PCR primers for use in the identification and detection of Actinobacillus actinomycetemcomitans. These primers target variable regions of the 16S ribosomal RNA coding gene (rDNA). We assessed the specificity of the primers against 9 A. actinomycetemcomitans strains and 11 strains (3 species) of the Haemophilus genus. Primer sensitivity was determined by testing serial dilutions of the purified genomic DNAs of A. actinomycetemcomitans ATCC 33384T. Our obtained data revealed that we had obtained species-specific amplicons for all of the tested A. actinomycetemcomitans strains, and that none of these amplicons occurred in any of the other species. Our PCR protocol proved able to detect as little as 4 fg of A. actinomycetemcomitans chromosomal DNA. Our findings suggest that these PCR primers are incredibly sensitive, and should prove suitable for application in epidemiological studies, as well as the diagnosis and monitoring of periodontal pathogens after treatment for periodontitis.
Kim
Su-Gwan
SG
Department of Oral and Maxillofacial Surgery, College of Dentistry, Chosun University. 375 Seo-Suk Dong, Dong-ku, Gwang-ju 501-759, Republic of Korea.
Kim
Soo-Heung
SH
Kim
Mi-Kwang
MK
Kim
Hwa-Sook
HS
Kook
Joong-Ki
JK
D016976:Aggregatibacter actinomycetemcomitans / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers; D004275:DNA, Ribosomal / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S