Isolation and identification of bacteria from the root canal of the teeth diagnosed as the acute pulpitis and acute periapical abscess

Article information

Restor Dent Endod. 2005;30(5):409-422

Publication date (electronic) : 2005 September 30

doi : https://doi.org/10.5395/JKACD.2005.30.5.409

Yeon-Jae Lee ¹, Mi-Kwang Kim ², Ho-Keel Hwang ¹^,³, Joong-Ki Kook ²^,³

¹Department of Conservative Dentistry, College of Dentistry, Chosun University, Korea.

²Department of Oral Biochemistry, College of Dentistry, Chosun University, Korea.

³Oral Biology Research Institute, College of Dentistry, Chosun University, Korea.

Corresponding author: Joong-Ki Kook. Department of Oral Biochemistry, and Oral Biology Research Institute, College of Dentistry, Chosun University, 375 Seo-suk Dong, Dong-Gu, Gwang-ju, 501-759 Korea. Tel: 82-62-230-6877, Fax: 82-62-224-3706, jkkook@chosun.ac.kr

Received 2005 April 11; Revised 2005 May 16; Accepted 2005 June 13.

Abstract

The aim of this study was to identify the bacteria isolated from acute endodontic lesions by cell culture and 16S rDNA sequencing. The necrotic pulpal tissue was collected from 17 infected root canals, which were diagnosed as being either an acute pulpitis or acute periapical abscess. Samples were collected aseptically from the infected pulpal tissue of the infected root canals using a barbed broach and a paper point. The cut barbed broaches and paper points were transferred to an eppendorf tube containing 500 ul of 1 X PBS. The sample solution was briefly mixed and plated onto a BHI-agar plate containing 5% sheep blood. The agar plates were incubated in a 37℃ anaerobic chamber for 7 days. The bacteria growing on the agar plate were identified by 16S rRNA coding gene (rDNA) cloning and sequencing at the species level. Among the 71 colonies grown on the agar plates, 56 strains survived and were identified. In dental caries involving the root canals, Streptococcus spp. were mainly isolated. Actinomyces, Clostridia, Bacteroides and Fusobacteria were isolated in the periapical lesion without dental caries. Interestingly, two new Actinomyces spp. (ChDC B639 and ChDC B631) were isolated in this study. These results showed that there was diversity among the species in endodontic lesions. This suggests that an endodontic infection is a mixed infection with a polymicrobial etiology. These results may offer the bacterial strains for pathogenesis studies related to an endodontic infection.

Keywords: Acute pulpitis; Acute periapical abscess; Bacteria; 16S rRNA gene

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