Loop-mediated isothermal amplification method for rapid detection of the periodontopathic bacteria Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola

Journal of Clinical Microbiology
Akihiro YoshidaTadamichi Takehara

Abstract

Loop-mediated isothermal amplification (LAMP), a novel nucleic acid amplification method, was developed for the rapid detection of the major periodontal pathogens Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola. The LAMP method amplifies DNA with high specificity, efficiency, and rapidity under isothermal conditions using a set of four specially designed primers and a DNA polymerase with strand displacement activity. In this study, we initially designed the primers for LAMP assays to detect these bacteria and evaluated the specificity and sensitivity of these assays. The specificities of the primers for these bacteria were examined using various oral bacteria and various reaction times. The lower detection limits of the 60-min LAMP reaction without loop primers were 1 microg/tube for P. gingivalis, 10 fg/tube for T. forsythia, and 1 ng/tube for T. denticola. Addition of the loop primers for each bacterium improved the detection specificities and sensitivities by several magnitudes. Furthermore, LAMP assays were applied to the rapid detection of these periodontal pathogens in clinical specimens, and the results were compared with those of conventional PCR detection. The results of the LAMP assays correspo...Continue Reading

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Citations

Feb 7, 2008·PLoS Neglected Tropical Diseases·Zablon Kithinji NjiruAndrew Richard Christopher Thompson
Nov 3, 2012·Journal of Oral Microbiology·Edward ShillitoeRobert Cooney
May 9, 2006·Gerodontology·Masayo TachibanaTadamichi Takehara
Jun 3, 2015·Jundishapur Journal of Microbiology·Parastoo ChamanrokhDavood Esmaili
Jun 21, 2008·FEMS Immunology and Medical Microbiology·Junko MiyagawaShogo Takashiba
Apr 26, 2017·Journal of Global Antimicrobial Resistance·Amornrat ThuengernKrit Thirapanmethee
Oct 27, 2017·Current Medicinal Chemistry·Agnieszka ChylewskaMariusz Makowski
Oct 5, 2019·PloS One·Diona PodushkinaEdward M Golenberg
Apr 4, 2021·Journal of Clinical Medicine·Marcin LenkowskiAnna Surdacka
Aug 28, 2021·Cells·Marianna SorokaAnna Rymaszewska

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