Development of species-specific quantitative real-time PCR primers for detecting Lautropia mirabilis
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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.
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.
참고문헌 (13)
- Gerner-Smidt P, "Lautropia mirabilis gen. nov., sp. nov., a gram-negative motile coccus with unusual morphology isolated from the human mouth", Microbiology (Reading), 140(Pt 7), pp.1787~1797 (1994)[DOI]
- Lim YK, "isolated from human dental plaque of a gingivitis lesion", Curr Microbiol, 76, pp.1369~1373 (2019)[DOI]
- Colombo AP, "Comparisons of subgingival microbial profiles of refractory periodontitis, severe periodontitis, and periodontal health using the human oral microbe identification microarray", J Periodontol, 80, pp.1421~1432 (2009)[DOI]
- Colombo AP, "Impact of periodontal therapy on the subgingival microbiota of severe periodontitis : comparison between good responders and individuals with refractory periodontitis using the human oral microbe identification microarray", J Periodontol, 83, pp.1279~1287 (2012)[DOI]
- Shaddox LM, "Microbiological characterization in children with aggressive periodontitis", J Dent Res, 91, pp.927~933 (2012)[DOI]
- Papapanou PN, "Subgingival microbiome and clinical periodontal status in an elderly cohort : the WHICAP ancillary study of oral health", J Periodontol, 91(Suppl 1), pp.S56~S67 (2020)[DOI]
- Al-Kamel A, "Subgingival microbiome of experimental gingivitis : shifts associated with the use of chlorhexidine and N-acetyl cysteine mouthwashes", J Oral Microbiol, 11, pp.1608141~ (2019)[DOI]
- Ben Dekhil SM, "Isolation of Lautropia mirabilis from sputa of a cystic fibrosis patient", J Clin Microbiol, 35, pp.1024~1026 (1997)[DOI]
- Tan RYP, "The first case of peritoneal dialysisassociated peritonitis due to Lautropia mirabilis", Nephrology(Carlton), 26, pp.634~635 (2021)[DOI]
- Severinov K, "Structural modules of the large subunits of RNA polymerase. Introducing archaebacterial and chloroplast split sites in the beta and beta’ subunits of Escherichia coli RNA polymerase", J Biol Chem, 271, pp.27969~27974 (1996)[DOI]
- Park SN, "Development of quantitative realtime PCR primers for detecting 42 oral bacterial species", Arch Microbiol, 195, pp.473~482 (2013)[DOI]
- Wilson K, "Preparation of genomic DNA from bacteria", Curr Protoc Mol Biol, (Chapter 2:Unit 2.4) (2001)[DOI]
- 박순낭, "Development of Porphyromonas gingivalis-Specific Quantitative Real-Time PCR Primers Based on the Nucleotide Sequence of rpoB", The Journal of Microbiology, 49(2), pp.315~319 (2011), 한국미생물학회