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KMID : 0848120210460030140
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International Journal of Oral Biology
2021 Volume.46 No. 3 p.140 ~ p.145
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Developing species-specific quantitative real-time polymerase chain reaction primers for detecting Lautropia mirabilis
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Park Soon-Nang
Kook Joong-Ki
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Abstract
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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.
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KEYWORD
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Lautropia mirabilis, 16S rDNA, Quantitative real-time polymerase chain reaction primers
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