다중 정량 PCR을 이용한 치수 감염 원인 병원성 세균의 동시 검출 및 신속 동정
Rapid Simultaneous Detection and Identification of Endodontic Pathogenic Bacteria by Multiplex Quantitative PCR
장우성, 김가연,
소속 상세정보
장우성 ( Chang Woo-Sung ) - 신구대학 치위생과
김가연 ( Kim Ga-Yeon ) - 단국대학교 보건과학대학 치위생학과
Abstract
The purpose of this study is to develop the quantitative PCR(qPCR) assay that would enable the rapid identification and simultaneous detection of six different endodontic pathogenic bacteria in a single reaction. In this study, six pairs of primers for Treponema denticola, Porphyromonas gingivalis, Fusobacterium nucleatum, Prevotella intermedia, Streptococcus mutans, and Staphylococcus aureus and two pairs of housekeeping genes were designed for a multiplex qPCR based on the SYBR Green method. The genomic DNA was extracted from reference strains and submitted to the qPCR reaction. The specificity of the amplified products was analyzed by melting curves. As a result, six distinct melting peaks were identified by the melting curve analysis and all of the target species were simultaneously discriminated. Therefore, the multiplex qPCR assay developed in this study can be used for rapid identification and detection of T. denticola, P. gingivalis, F. nucleatum, P. intermedia, S. mutans, and S. aureus at the same time. In combination with the melting curve analysis, the level of the target species and total bacterial load can be obtained.
키워드
Bacterial detection; Diagnostic identification; Endodontic infection; Multiplex quantitative PCR
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