Loop-mediated isothermal amplification combined with PCR and immunohistochemistry for detecting Porphyromonas gingivalis in periapical periodontitis
Porphyromonas gingivalis is important in the development of marginal periodontitis. However, the precise role and localization of P. gingivalis in chronic periapical periodontitis remain unclear. Thus, methods that can detect P. gingivalis in formalin-fixed and paraffin-embedded (FFPE) tissue sample...
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| Veröffentlicht in: | Journal of Oral Science 2016, Vol.58(2), pp.163-169 |
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| creator | Kitano, Taiichi Mikami, Yoshikazu Iwase, Takashi Asano, Masatake Komiyama, Kazuo |
| description | Porphyromonas gingivalis is important in the development of marginal periodontitis. However, the precise role and localization of P. gingivalis in chronic periapical periodontitis remain unclear. Thus, methods that can detect P. gingivalis in formalin-fixed and paraffin-embedded (FFPE) tissue samples are needed. We assessed a technique combining loop-mediated isothermal amplification (LAMP) with PCR (PCR-LAMP) for detection of P. gingivalis, using 110 FFPE tissue samples of chronic apical periodontitis. PCR-LAMP specifically detected P. gingivalis with high sensitivity in FFPE tissue samples, and the sensitivity of the technique was higher than that of PCR or LAMP alone. The results of immunohistochemistry (IHC) confirmed the specificity of PCR-LAMP. IHC showed that P. gingivalis was localized in a granular layer of chronic apical periodontitis, a region that correlated with the localization of macrophages. This is the first study to describe the localization of P. gingivalis in human periapical periodontitis. In conclusion, PCR-LAMP was an effective tool for detecting P. gingivalis in periapical periodontitis. In addition, IHC results improve our understanding of the role of P. gingivalis in the progression of periapical periodontitis. (J Oral Sci 58, 163-169, 2016) |
| doi_str_mv | 10.2334/josnusd.15-0665 |
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However, the precise role and localization of P. gingivalis in chronic periapical periodontitis remain unclear. Thus, methods that can detect P. gingivalis in formalin-fixed and paraffin-embedded (FFPE) tissue samples are needed. We assessed a technique combining loop-mediated isothermal amplification (LAMP) with PCR (PCR-LAMP) for detection of P. gingivalis, using 110 FFPE tissue samples of chronic apical periodontitis. PCR-LAMP specifically detected P. gingivalis with high sensitivity in FFPE tissue samples, and the sensitivity of the technique was higher than that of PCR or LAMP alone. The results of immunohistochemistry (IHC) confirmed the specificity of PCR-LAMP. IHC showed that P. gingivalis was localized in a granular layer of chronic apical periodontitis, a region that correlated with the localization of macrophages. This is the first study to describe the localization of P. gingivalis in human periapical periodontitis. In conclusion, PCR-LAMP was an effective tool for detecting P. gingivalis in periapical periodontitis. In addition, IHC results improve our understanding of the role of P. gingivalis in the progression of periapical periodontitis. (J Oral Sci 58, 163-169, 2016)</description><identifier>ISSN: 1343-4934</identifier><identifier>EISSN: 1880-4926</identifier><identifier>DOI: 10.2334/josnusd.15-0665</identifier><identifier>PMID: 27349536</identifier><language>eng</language><publisher>Japan: Nihon University School of Dentistry</publisher><subject>Dentistry ; Humans ; Immunohistochemistry ; LAMP ; Limit of Detection ; PCR ; periapical periodontitis ; Periapical Periodontitis - microbiology ; Polymerase Chain Reaction - methods ; Porphyromonas gingivalis ; Porphyromonas gingivalis - isolation & purification</subject><ispartof>Journal of Oral Science, 2016, Vol.58(2), pp.163-169</ispartof><rights>2016 by Nihon University School of Dentistry</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c560t-abc9c5e44fcc687b96480a8a8a60c5ceb8c3fa0b72237b2fd2731e84343fd4be3</citedby><cites>FETCH-LOGICAL-c560t-abc9c5e44fcc687b96480a8a8a60c5ceb8c3fa0b72237b2fd2731e84343fd4be3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,1877,4010,27900,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27349536$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kitano, Taiichi</creatorcontrib><creatorcontrib>Mikami, Yoshikazu</creatorcontrib><creatorcontrib>Iwase, Takashi</creatorcontrib><creatorcontrib>Asano, Masatake</creatorcontrib><creatorcontrib>Komiyama, Kazuo</creatorcontrib><title>Loop-mediated isothermal amplification combined with PCR and immunohistochemistry for detecting Porphyromonas gingivalis in periapical periodontitis</title><title>Journal of Oral Science</title><addtitle>J Oral Sci</addtitle><description>Porphyromonas gingivalis is important in the development of marginal periodontitis. However, the precise role and localization of P. gingivalis in chronic periapical periodontitis remain unclear. Thus, methods that can detect P. gingivalis in formalin-fixed and paraffin-embedded (FFPE) tissue samples are needed. We assessed a technique combining loop-mediated isothermal amplification (LAMP) with PCR (PCR-LAMP) for detection of P. gingivalis, using 110 FFPE tissue samples of chronic apical periodontitis. PCR-LAMP specifically detected P. gingivalis with high sensitivity in FFPE tissue samples, and the sensitivity of the technique was higher than that of PCR or LAMP alone. The results of immunohistochemistry (IHC) confirmed the specificity of PCR-LAMP. IHC showed that P. gingivalis was localized in a granular layer of chronic apical periodontitis, a region that correlated with the localization of macrophages. This is the first study to describe the localization of P. gingivalis in human periapical periodontitis. In conclusion, PCR-LAMP was an effective tool for detecting P. gingivalis in periapical periodontitis. In addition, IHC results improve our understanding of the role of P. gingivalis in the progression of periapical periodontitis. (J Oral Sci 58, 163-169, 2016)</description><subject>Dentistry</subject><subject>Humans</subject><subject>Immunohistochemistry</subject><subject>LAMP</subject><subject>Limit of Detection</subject><subject>PCR</subject><subject>periapical periodontitis</subject><subject>Periapical Periodontitis - microbiology</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Porphyromonas gingivalis</subject><subject>Porphyromonas gingivalis - isolation & purification</subject><issn>1343-4934</issn><issn>1880-4926</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kUuPFCEUhYnROOPo2p1h6aZmoHgUvTSd8ZF04sTomlDUrS46BZRAafp_-IOl7bZDwj2Bj0PuuQi9peS-ZYw_HGIOax7uqWiIlOIZuqVKkYZvWvm8asZZ1YzfoFc5HwjhrezES3TTdoxvBJO36M8uxqXxMDhTYMAuxzJB8mbGxi-zG501xcWAbfS9C5X47cqEn7bfsAkV934NcXK5RDuBrzUd8RgTHqCALS7s8VNMy3RM0cdgMt7XI_fLzC5jF_ACyZmlfjH_k3GIobji8mv0YjRzhjeXeod-fHz8vv3c7L5--rL9sGuskKQ0prcbK4Dz0Vqpun4juSJG1SWJFRZ6ZdloSN-1Lev6dhxq2xQUr6mMA--B3aH3Z98lxZ8r5KJrCxbm2QSIa9ZU1cgIJVRW9OGM2hRzTjDqJTlv0lFTok-j0JdRaCr0aRT1xbuL-drXfK_8_-wr8HgGDrmYPVwBk4qzM1wNhdLtabsYX-_tZJKGwP4CGBylcQ</recordid><startdate>2016</startdate><enddate>2016</enddate><creator>Kitano, Taiichi</creator><creator>Mikami, Yoshikazu</creator><creator>Iwase, Takashi</creator><creator>Asano, Masatake</creator><creator>Komiyama, Kazuo</creator><general>Nihon University School of Dentistry</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>2016</creationdate><title>Loop-mediated isothermal amplification combined with PCR and immunohistochemistry for detecting Porphyromonas gingivalis in periapical periodontitis</title><author>Kitano, Taiichi ; Mikami, Yoshikazu ; Iwase, Takashi ; Asano, Masatake ; Komiyama, Kazuo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c560t-abc9c5e44fcc687b96480a8a8a60c5ceb8c3fa0b72237b2fd2731e84343fd4be3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Dentistry</topic><topic>Humans</topic><topic>Immunohistochemistry</topic><topic>LAMP</topic><topic>Limit of Detection</topic><topic>PCR</topic><topic>periapical periodontitis</topic><topic>Periapical Periodontitis - microbiology</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Porphyromonas gingivalis</topic><topic>Porphyromonas gingivalis - isolation & purification</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kitano, Taiichi</creatorcontrib><creatorcontrib>Mikami, Yoshikazu</creatorcontrib><creatorcontrib>Iwase, Takashi</creatorcontrib><creatorcontrib>Asano, Masatake</creatorcontrib><creatorcontrib>Komiyama, Kazuo</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of Oral Science</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kitano, Taiichi</au><au>Mikami, Yoshikazu</au><au>Iwase, Takashi</au><au>Asano, Masatake</au><au>Komiyama, Kazuo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Loop-mediated isothermal amplification combined with PCR and immunohistochemistry for detecting Porphyromonas gingivalis in periapical periodontitis</atitle><jtitle>Journal of Oral Science</jtitle><addtitle>J Oral Sci</addtitle><date>2016</date><risdate>2016</risdate><volume>58</volume><issue>2</issue><spage>163</spage><epage>169</epage><pages>163-169</pages><issn>1343-4934</issn><eissn>1880-4926</eissn><abstract>Porphyromonas gingivalis is important in the development of marginal periodontitis. However, the precise role and localization of P. gingivalis in chronic periapical periodontitis remain unclear. Thus, methods that can detect P. gingivalis in formalin-fixed and paraffin-embedded (FFPE) tissue samples are needed. We assessed a technique combining loop-mediated isothermal amplification (LAMP) with PCR (PCR-LAMP) for detection of P. gingivalis, using 110 FFPE tissue samples of chronic apical periodontitis. PCR-LAMP specifically detected P. gingivalis with high sensitivity in FFPE tissue samples, and the sensitivity of the technique was higher than that of PCR or LAMP alone. The results of immunohistochemistry (IHC) confirmed the specificity of PCR-LAMP. IHC showed that P. gingivalis was localized in a granular layer of chronic apical periodontitis, a region that correlated with the localization of macrophages. This is the first study to describe the localization of P. gingivalis in human periapical periodontitis. In conclusion, PCR-LAMP was an effective tool for detecting P. gingivalis in periapical periodontitis. In addition, IHC results improve our understanding of the role of P. gingivalis in the progression of periapical periodontitis. (J Oral Sci 58, 163-169, 2016)</abstract><cop>Japan</cop><pub>Nihon University School of Dentistry</pub><pmid>27349536</pmid><doi>10.2334/josnusd.15-0665</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; J-STAGE (Japan Science & Technology Information Aggregator, Electronic) Freely Available Titles - Japanese; Open Access Titles of Japan; EZB-FREE-00999 freely available EZB journals |
| subjects | Dentistry Humans Immunohistochemistry LAMP Limit of Detection PCR periapical periodontitis Periapical Periodontitis - microbiology Polymerase Chain Reaction - methods Porphyromonas gingivalis Porphyromonas gingivalis - isolation & purification |
| title | Loop-mediated isothermal amplification combined with PCR and immunohistochemistry for detecting Porphyromonas gingivalis in periapical periodontitis |
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