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Journal of Clinical Microbiology, September 2002, p. 3223-3231, Vol. 40, No. 9
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.9.3223-3231.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

PCR-Based Identification of Bacteria Associated with Endodontic Infections

Ashraf F. Fouad,1* Jody Barry,1 Melissa Caimano,2 Michael Clawson,3 Qiang Zhu,1 Rachaele Carver,1 Karsten Hazlett,4 and Justin D. Radolf4

Department of Endodontology, School of Dental Medicine,1 Department of Pathology,2 Center for Microbial Pathogenesis, University of Connecticut Health Center, Farmington, Connecticut,4 U.S. Meat Animal Research Center, Agricultural Research Service, U.S. Department of Agriculture, Clay Center, Nebraska3

Received 26 November 2001/ Returned for modification 15 February 2002/ Accepted 10 June 2002

PCR primers that target the bacterial 16S rRNA genes (or the tuf gene for the genus Enterococcus) were used to identify 10 putative bacterial pathogens in root canals with necrotic pulp. In addition, the associations of these microorganisms with symptoms and a history of diabetes mellitus were investigated. Microbial samples from the root canals of 24 teeth with necrotic pulp were included in the study. PCR with universal bacterial primers identified bacterial DNA in 22 specimens; the remaining 2 specimens were from intact teeth that had been traumatized 6 months prior to treatment. PCR with specific primers showed that preoperative symptoms were significantly associated with the presence of Streptococcus spp. (P < 0.001 by chi-square analysis). There was also a nonsignificant trend for symptoms to be associated with Fusobacterium nucleatum and Porphyromonas gingivalis (odds ratio, >2) and for diabetes mellitus to be associated with P. gingivalis and Porphyromonas endodontalis (odds ratio, >2). Cloning and sequencing of the universal PCR product in one specimen revealed the presence of an organism related to the genus Olsenella, which has not previously been described in endodontic infections.

* Corresponding author. Mailing address: Department of Endodontology, University of Connecticut Health Center, 263 Farmington Ave., Farmington, CT 06030-1715. Phone: (860) 679-2726. Fax: (860) 679-2208. E-mail: fouad{at}nso.uchc.edu.

Journal of Clinical Microbiology, September 2002, p. 3223-3231, Vol. 40, No. 9
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.9.3223-3231.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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