細菌１６ＳリボソームＲＮＡ遺伝子に基づくＤＮＡプローブを用いた歯周病細菌の同定　　コロニーハイブリダイゼーション法

DOI 被引用文献4件 参考文献31件

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タイトル別名
  • Identification of Periodontal Bacteria Using DNA Probes Based on Bacterial 16S Ribosomal RNA Gene. Colony Hybridization Method.
  • olony Hybridization Method. -
  • - コロニーハイブリダイゼーション法-

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抄録

A new method to detect periodontal bacteria was developed by adapting colony hybridization using DNA probes. The synthetic oligonucleotides complementary to species-specific sequence in the 16S ribosomal RNA (rRNA) gene were designed for the detection of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia. The specificity of each DNA probe was verified by Northern hybridization between the 32Plabeled probe and bacterial RNAs. Each probe gave an intense signal specifically against DNAs derived from the corresponding bacterial strain including clinical isolates as low as 104 cells. The RNAs of bacterial colonies lifted from agar plates to nylon membranes were also detected by hybridization with these probes. The colony hybridization method combined with the cultivation on selective media allowed the detect of A. actinomycetemcomitans, P. gingivalis and P. intermedia from the dental plaques obtained from periodontitis patients. Detection of the bacteria by routine culture and biochemical test is time-consuming and very complicated. This method is simple and makes detecion and quantification of bacterial species easy, furthermore, it is a useful tool for analyzing the microbial flora from periodontal pockets.

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