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http://dx.doi.org/10.11620/IJOB.2021.46.3.140

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

Park, Soon-Nang (Korean Collection for Oral Microbiology, College of Dentistry, Chosun University)
Kook, Joong-Ki (Korean Collection for Oral Microbiology, College of Dentistry, Chosun University)
Publication Information
International Journal of Oral Biology / v.46, no.3, 2021 , pp. 140-145 More about this Journal
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.
Keywords
Lautropia mirabilis; 16S rDNA; Quantitative real-time polymerase chain reaction primers;
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1 Lim YK, Park SN, Lee WP, Shin JH, Jo E, Shin Y, Paek J, Chang YH, Kim H, Kook JK. Lautropia dentalis sp. nov., isolated from human dental plaque of a gingivitis lesion. Curr Microbiol 2019;76:1369-73. doi: 10.1007/s00284-019-01761-1.   DOI
2 Colombo AP, Boches SK, Cotton SL, Goodson JM, Kent R, Haffajee AD, Socransky SS, Hasturk H, Van Dyke TE, Dewhirst F, Paster BJ. Comparisons of subgingival microbial profiles of refractory periodontitis, severe periodontitis, and periodontal health using the human oral microbe identification microarray. J Periodontol 2009;80:1421-32. doi: 10.1902/jop.2009.090185.   DOI
3 Colombo AP, Bennet S, Cotton SL, Goodson JM, Kent R, Haffajee AD, Socransky SS, Hasturk H, Van Dyke TE, Dewhirst FE, Paster BJ. 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 2012;83:1279-87. doi: 10.1902/jop.2012.110566.   DOI
4 Shaddox LM, Huang H, Lin T, Hou W, Harrison PL, Aukhil I, Walker CB, Klepac-Ceraj V, Paster BJ. Microbiological characterization in children with aggressive periodontitis. J Dent Res 2012;91:927-33. doi: 10.1177/0022034512456039.   DOI
5 Park SN, Park JY, Kook JK. Development of Porphyromonas gingivalis-specific quantitative real-time PCR primers based on the nucleotide sequence of rpoB. J Microbiol 2011;49:315-9. doi: 10.1007/s12275-011-1028-y.   DOI
6 Severinov K, Mustaev A, Kukarin A, Muzzin O, Bass I, Darst SA, Goldfarb A. 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 1996;271:27969-74. doi: 10.1074/jbc.271.44.27969.   DOI
7 Papapanou PN, Park H, Cheng B, Kokaras A, Paster B, Burkett S, Watson CW, Annavajhala MK, Uhlemann AC, Noble JM. Subgingival microbiome and clinical periodontal status in an elderly cohort: the WHICAP ancillary study of oral health. J Periodontol 2020;91 Suppl 1:S56-67. doi: 10.1002/JPER.20-0194.   DOI
8 Al-Kamel A, Baraniya D, Al-Hajj WA, Halboub E, Abdulrab S, Chen T, Al-Hebshi NN. Subgingival microbiome of experimental gingivitis: shifts associated with the use of chlorhexidine and N-acetyl cysteine mouthwashes. J Oral Microbiol 2019;11:1608141. doi: 10.1080/20002297.2019.1608141.   DOI
9 Ben Dekhil SM, Peel MM, Lennox VA, Stackebrandt E, Sly LI. Isolation of Lautropia mirabilis from sputa of a cystic fibrosis patient. J Clin Microbiol 1997;35:1024-6. doi: 10.1128/jcm.35.4.1024-1026.1997.   DOI
10 Tan RYP, Tan BQ, Rao N. The first case of peritoneal dialysisassociated peritonitis due to Lautropia mirabilis. Nephrology (Carlton) 2021;26:634-5. doi: 10.1111/nep.13866.   DOI
11 Gerner-Smidt P, Keiser-Nielsen H, Dorsch M, Stackebrandt E, Ursing J, Blom J, Christensen AC, Christensen JJ, Frederiksen W, Hoffmann S, Holten-Andersen W, Ying YT. Lautropia mirabilis gen. nov., sp. nov., a gram-negative motile coccus with unusual morphology isolated from the human mouth. Microbiology (Reading) 1994;140(Pt 7):1787-97. doi: 10.1099/13500872-140-7-1787.   DOI
12 Wilson K. Preparation of genomic DNA from bacteria. Curr Protoc Mol Biol 2001;Chapter 2:Unit 2.4. doi: 10.1002/0471142727.mb0204s56.   DOI
13 Park SN, Lim YK, Kook JK. Development of quantitative realtime PCR primers for detecting 42 oral bacterial species. Arch Microbiol 2013;195:473-82. doi: 10.1007/s00203-013-0896-4.   DOI