DNA 프로브 Pig27 핵산염기서열을 기반으로 한 Prevotella intermedia의 검출을 위한 PCR 프라이머 개발

Abstract

In a previous study, a Prevotella intermedia (P. intermedia)-specific DNA probe, Pig27, was cloned by shotgun method and southern blot hybridization. The purpose of this study was to develop the P. intermedia species-specific PCR primers based on the nucleotide sequence of Pig27 DNA probe. The specificity of the PCR primers was tested against 6 clinical isolates of P. intermedia, 10 clinical isolates of P. nigrescens and 20 type strains of oral bacteria. The sensitivity of PCR primers was determined by testing 0.4 ng～4 fg of the purified genomic DNA. The data showed that the two sets of PCR primers, Pig27-F2/Pig27-R2 and Pig27-F5/Pig27-R5 had the species-specificity for the P. intermedia ATCC 25611T. The detection limits of four primer sets was 0.4 pg～4 pg of the purified genomic DNA of P. intermedia ATCC 25611T. These results suggested that the two sets of PCR primer, Pig27-F2/Pig27-R2 and Pig27-F5/Pig27-R5, could be useful in the detection kit of P. intermedia ATCC 25611T.