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Developing species-specific quantitative real-time polymerase chain reaction primers for detecting Lautropia mirabilis KCI 등재후보

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  • URLhttps://db.koreascholar.com/Article/Detail/409738
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대한구강생물학회 (The Korean Academy of Oral Biology)
초록

This study aimed to develop Lautropia mirabilis -specific quantitative real-time polymerase chain reaction (qPCR) primers based on the sequence of DNA-directed RNA polymerase subunit beta gene. The PrimerSelect program was used in designing of the qPCR primers, RTLam-F4 and RTLam-R3. The specificity of the qPCR primers were performed by conventional PCR with 37 strains of 37 oral bacterial species, including L. mirabilis . The sensitivity of the primers was determined by qPCR with the serial dilution of purified genomic DNA of L. mirabilis KCOM 3484, ranged from 4 ng to 4 fg. The data showed that the qPCR primers could detect only L. mirabilis strains and as little as 40 fg of genome DNA of L. mirabilis KCOM 3484. These results indicate that this qPCR primer pair (RTLam-F4/ RTLam-R3) may be useful for species-specific detection of L. mirabilis in epidemiological studies of oral bacterial infectious diseases such as periodontal disease.

목차
Introduction
Materials and Methods
    1. 세균 및 세균 배양
    2. qPCR 프라이머 설계
    3. 세균 지놈 DNA 추출
    4. 온도 구배(gradient) PCR 및 qPCR 프라이머 쌍의종-특이성 검증
    5. qPCR을 이용한 민감도(검출 한계) 조사
Results
Discussion
References
저자
  • Soon-Nang Park(Korean Collection for Oral Microbiology, College of Dentistry, Chosun University/Department of Oral Biochemistry, College of Dentistry, Chosun University)
  • Joong-Ki Kook(Korean Collection for Oral Microbiology, College of Dentistry, Chosun University/Department of Oral Biochemistry, College of Dentistry, Chosun University) Correspondence to