Direct detection of Actinomyces spp. from infected root canals in a Chinese population: a study using PCR-based, oligonucleotide-DNA hybridization technique.
OBJECTIVE
The poor sensitivity of phenotypic identification techniques has hampered the taxonomic differentiation of Actinomyces. Hence we developed a sensitive and specific, PCR-based oligonucleotide-DNA hybridization technique to detect Actinomyces spp. and, used this method to detect these organisms in samples directly obtained from infected root canals.


METHODS
A total of 32 samples from 28 Chinese patients, with primary root canal infections, aseptically exposed at the first patient visit, were studied. Whole bacterial genomic DNA was isolated directly from paper point samples. The variable regions of 16S ribosomal DNA of bacteria were amplified and labeled with digoxigenin for further hybridization and detection. A total of seven oligonucleotide probes specific for A. bovis, A. gerencseriae, A. israelii, A. meyeri, catalase-negative A. naeslundii (genospecies 1 and 2), catalase-positive A. naeslundii genospecies 2 and A. odontolyticus were used.


RESULTS
16 of the 32 teeth were infected with one or more Actinomyces species. The prevalence rates of the examined species were: A. odontolyticus 31.3%, A. meyeri 9.4%, A. naeslundii 9.4%, A. israelii 6.3% and A. gerencseriae 3.1%; no A. bovis was detected in any of the canals. Furthermore, A. odontolyticus was isolated more frequently from root canals with caries or a history of caries (Fisher's exact test: P=0.0496; Odds ratio=9.00, 95% confidence interval: 0.97-83.63), and A. naeslundii was significantly associated with traumatized teeth (Fisher's exact test: P=0.0121; Odds ratio=57.00, 95% confidence interval: 2.10-1546.90). However, no significant correlation was found between Actinomyces spp. and clinical symptoms and signs, such as pain, swelling, percussion to tenderness, sinus and periapical radiolucency.


CONCLUSIONS
Actinomyces spp. may be important pathogens of root canal infections. A. naeslundii in particular may be related with traumatized teeth. A. odontolyticus appears to be involved in infections related to caries, exposure of dentinal tubules during cavity preparation and/or leaking restoration, but further clarification with large samples is necessary.
Tang
Gaoyan
G
Oral Bio-Sciences, Faculty of Dentistry, The University of Hong Kong, Hong Kong SAR, China.
Samaranayake
Lakshman P
LP
Yip
Hak-Kong
HK
Chu
Frederick C S
FC
Tsang
Peter C S
PC
Cheung
Becky P K
BP
D000190:Actinomyces / Q000145:classification* / Q000302:isolation & purification; D000196:Actinomycosis / Q000382:microbiology*; D000293:Adolescent; D000328:Adult; D000368:Aged; D002681:China; D016001:Confidence Intervals; D004269:DNA, Bacterial / Q000032:analysis; D003731:Dental Caries / Q000382:microbiology; D003786:Dental Pulp Cavity / Q000382:microbiology*; D003788:Dental Pulp Diseases / Q000382:microbiology*; D005260:Female; D005838:Genotype; D006801:Humans; D008297:Male; D008875:Middle Aged; D009693:Nucleic Acid Hybridization; D016017:Odds Ratio; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D018677:Tooth Injuries / Q000382:microbiology
D004269:DNA, Bacterial; D015345:Oligonucleotide Probes; D012336:RNA, Ribosomal, 16S