Prevotella intermedia 동정에 이용될 Pig6 DNA 프로브의 특이성 평가

Abstract

The purpose of this study is to develop the strain-specific PCR primers for the identification of Prevotella intermedia G8-9K-3 which is frequently used in the pathogenesis studies of periodontitis. The HindIII-digested genomic DNA of P. intermedia G8-9K-3 were cloned by random cloning method. The specificity of cloned DNA fragments were determined by Southern blot analysis. The nucleotide sequence of cloned DNA probes was determined by chain termination method. The PCR primers were designed based on the nucleotide sequence of cloned DNA fragment. The data showed that Pig6 DNA probe were hybridized with the genomic DNA from P. intermedia strains (ATCC 25611 and G8-9K-3) isolated from the Westerns, not the strains isolated from Koreans. The Pig6 DNA probe were consisted of 813 bp. Pig6-F3 and Pig6-R3 primers, designed base on the nucleotide sequences of Pig6 DNA probe, were also specific to the only both P. intermedia ATCC 25611 and P. intermedia G8-9K-3. In the other hand, Pig6-F7 and Pig6-R7 primers were specific to the only P. intermedia G8-9K-3. These results indicate that Pig6-F7 and Pig6-R7 primers have proven useful for the identification of P. intermedia G8-9K-3, especially with regard to the maintenance of the strain.