comprehensive analysis of secondary dental root canal infections a combination of culture and culture-independent approaches reveals new insights二次牙齿根管感染的综合分析相结合的文化和文化无关的方法揭示了新见解.pdfVIP

Comprehensive Analysis of Secondary Dental Root Canal Infections: A Combination of Culture and Culture- Independent Approaches Reveals New Insights 1 1 1 2 3 Annette Carola Anderson , Elmar Hellwig , Robin Vespermann , Annette Wittmer , Michael Schmid , 1 1 Lamprini Karygianni , Ali Al-Ahmad * 1 Department of Operative Dentistry and Periodontology, Albert-Ludwigs-University, Freiburg, Germany, 2 Department of Hygiene and Microbiology, Albert-Ludwigs- ¨ University, Freiburg, Germany, 3 Michael Schmid, Research Unit Microbe-Plant Interactions, Helmholtz Zentrum Munchen, German Research Center for Environmental Health, Neuherberg, Germany Abstract Persistence of microorganisms or reinfections are the main reasons for failure of root canal therapy. Very few studies to date have included culture-independent methods to assess the microbiota, including non-cultivable microorganisms. The aim of this study was to combine culture methods with culture-independent cloning methods to analyze the microbial flora of root-filled teeth with periradicular lesions. Twenty-one samples from previously root-filled teeth were collected from patients with periradicular lesions. Microorganisms were cultivated, isolated and biochemically identified. In addition, ribosomal DNA of bacteria, fungi and archaea derived from the same samples was amplified and the PCR products were used to construct clone libraries. DNA of selected clones was sequenced and microbial species were identified, comparing the sequences with public databases. Microorganisms were found in 12