Developing species-specific quantitative real-time polymerase chain reaction primers for detecting Lautropia mirabilis
박순낭, 국중기,
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박순낭 ( Park Soon-Nang ) - Chosun University College of Dentistry Korean Collection for Oral Microbiology
국중기 ( Kook Joong-Ki ) - Chosun University College of Dentistry Korean Collection for Oral Microbiology
Abstract
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.
키워드
Lautropia mirabilis; 16S rDNA; Quantitative real-time polymerase chain reaction primers
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