<?xml version="1.0" encoding="ISO-8859-1"?><cms:container xmlns:cms="http://edoc.hu-berlin.de/diml/module/cms"><cms:document><cms:meta><cms:entry ref="front" type="front"/><cms:entry ref="_Toc119564310" type="link"/><cms:entry type="title">Prevalence analysis of putative periodontal pathogens in patients with aggressive periodontitis and healthy elderly. A molecular study</cms:entry><cms:entry type="author">Lilian  Edesi-Neuss</cms:entry><cms:entry ref="_Toc119564311" type="link"/><cms:entry ref="_Toc119564312" type="link"/><cms:entry id="chapter1" part="chapter1" ref="chapter1" type="chapter">1</cms:entry><cms:entry id="_Toc119564314" part="chapter1" ref="_Toc119564314" type="link"/><cms:entry id="N1012B" part="chapter1" ref="N1012B" type="citenumber">1</cms:entry><cms:entry id="N10135" part="chapter1" ref="N10135" type="section">1.1</cms:entry><cms:entry id="_Toc119564315" part="chapter1" ref="_Toc119564315" type="link"/><cms:entry id="N1013F" part="chapter1" ref="N1013F" type="citenumber">2</cms:entry><cms:entry id="_Toc119564316" part="chapter1" ref="_Toc119564316" type="link"/><cms:entry id="N101A3" part="chapter1" ref="N101A3" type="section">1.2</cms:entry><cms:entry id="N101AA" part="chapter1" ref="N101AA" type="citenumber">3</cms:entry><cms:entry id="N101CB" part="chapter1" ref="N101CB" type="citenumber">4</cms:entry><cms:entry id="N101D9" part="chapter1" ref="N101D9" type="citenumber">5</cms:entry><cms:entry id="N101E5" part="chapter1" ref="N101E5" type="citenumber">6</cms:entry><cms:entry id="N1020D" part="chapter1" ref="N1020D" type="subsection">1.2.1</cms:entry><cms:entry id="_Toc119564317" part="chapter1" ref="_Toc119564317" type="link"/><cms:entry id="_Toc119564318" part="chapter1" ref="_Toc119564318" type="link"/><cms:entry id="N1021F" part="chapter1" ref="N1021F" type="subsection">1.2.2</cms:entry><cms:entry id="N10226" part="chapter1" ref="N10226" type="citenumber">7</cms:entry><cms:entry id="_Toc119564319" part="chapter1" ref="_Toc119564319" type="link"/><cms:entry id="N10237" part="chapter1" ref="N10237" type="subsection">1.2.3</cms:entry><cms:entry id="N10248" part="chapter1" ref="N10248" type="table"/><cms:entry id="N10385" part="chapter1" ref="N10385" type="citenumber">8</cms:entry><cms:entry id="_Toc119564320" part="chapter1" ref="_Toc119564320" type="link"/><cms:entry id="N103A2" part="chapter1" ref="N103A2" type="subsection">1.2.4</cms:entry><cms:entry id="N103AC" part="chapter1" ref="N103AC" type="citenumber">9</cms:entry><cms:entry id="_Toc119564321" part="chapter1" ref="_Toc119564321" type="link"/><cms:entry id="N103EE" part="chapter1" ref="N103EE" type="subsection">1.2.5</cms:entry><cms:entry id="_Toc119564322" part="chapter1" ref="_Toc119564322" type="link"/><cms:entry id="N103FF" part="chapter1" ref="N103FF" type="subsection">1.2.6</cms:entry><cms:entry id="N10406" part="chapter1" ref="N10406" type="citenumber">10</cms:entry><cms:entry id="_Toc119564323" part="chapter1" ref="_Toc119564323" type="link"/><cms:entry id="N104A0" part="chapter1" ref="N104A0" type="section">1.3</cms:entry><cms:entry id="N104BC" part="chapter1" ref="N104BC" type="citenumber">11</cms:entry><cms:entry id="_Toc119564324" part="chapter1" ref="_Toc119564324" type="link"/><cms:entry id="N104D1" part="chapter1" ref="N104D1" type="section">1.4</cms:entry><cms:entry id="N104D8" part="chapter1" ref="N104D8" type="citenumber">12</cms:entry><cms:entry id="N104DE" part="chapter1" ref="N104DE" type="subsection">1.4.1</cms:entry><cms:entry id="_Toc119564325" part="chapter1" ref="_Toc119564325" type="link"/><cms:entry id="_Toc119564326" part="chapter1" ref="_Toc119564326" type="link"/><cms:entry id="N10502" part="chapter1" ref="N10502" type="subsection">1.4.2</cms:entry><cms:entry id="_Toc119564327" part="chapter1" ref="_Toc119564327" type="link"/><cms:entry id="N1053F" part="chapter1" ref="N1053F" type="subsection">1.4.3</cms:entry><cms:entry id="N10546" part="chapter1" ref="N10546" type="citenumber">13</cms:entry><cms:entry id="_Toc119564328" part="chapter1" ref="_Toc119564328" type="link"/><cms:entry id="N1055A" part="chapter1" ref="N1055A" type="subsection">1.4.4</cms:entry><cms:entry id="_Toc119564329" part="chapter1" ref="_Toc119564329" type="link"/><cms:entry id="N10586" part="chapter1" ref="N10586" type="section">1.5</cms:entry><cms:entry id="_Toc119564330" part="chapter1" ref="_Toc119564330" type="link"/><cms:entry id="N1059D" part="chapter1" ref="N1059D" type="section">1.6</cms:entry><cms:entry id="N105A2" part="chapter1" ref="N105A2" type="subsection">1.6.1</cms:entry><cms:entry id="_Toc119564331" part="chapter1" ref="_Toc119564331" type="link"/><cms:entry id="N105AC" part="chapter1" ref="N105AC" type="citenumber">14</cms:entry><cms:entry id="N105CF" part="chapter1" ref="N105CF" type="citenumber">15</cms:entry><cms:entry id="N10617" part="chapter1" ref="N10617" type="citenumber">16</cms:entry><cms:entry id="_Toc119564332" part="chapter1" ref="_Toc119564332" type="link"/><cms:entry id="N10646" part="chapter1" ref="N10646" type="subsection">1.6.2</cms:entry><cms:entry id="_Toc119564333" part="chapter1" ref="_Toc119564333" type="link"/><cms:entry id="N10677" part="chapter1" ref="N10677" type="section">1.7</cms:entry><cms:entry id="N1067E" part="chapter1" ref="N1067E" type="citenumber">17</cms:entry><cms:entry id="chapter2" part="chapter2" ref="chapter2" type="chapter">2</cms:entry><cms:entry id="_Toc119564334" part="chapter2" ref="_Toc119564334" type="link"/><cms:entry id="N1068E" part="chapter2" ref="N1068E" type="helpercitenumber">17</cms:entry><cms:entry id="chapter3" part="chapter3" ref="chapter3" type="chapter">3</cms:entry><cms:entry id="_Toc119564335" part="chapter3" ref="_Toc119564335" type="link"/><cms:entry id="N106C2" part="chapter3" ref="N106C2" type="section">3.1</cms:entry><cms:entry id="_Toc119564336" part="chapter3" ref="_Toc119564336" type="link"/><cms:entry id="N106CA" part="chapter3" ref="N106CA" type="helpercitenumber">17</cms:entry><cms:entry id="N106CF" part="chapter3" ref="N106CF" type="citenumber">18</cms:entry><cms:entry id="_Toc119564337" part="chapter3" ref="_Toc119564337" type="link"/><cms:entry id="N106FA" part="chapter3" ref="N106FA" type="section">3.2</cms:entry><cms:entry id="N10704" part="chapter3" ref="N10704" type="citenumber">19</cms:entry><cms:entry id="N10764" part="chapter3" ref="N10764" type="citenumber">20</cms:entry><cms:entry id="N10775" part="chapter3" ref="N10775" type="citenumber">21</cms:entry><cms:entry id="_Toc119564338" part="chapter3" ref="_Toc119564338" type="link"/><cms:entry id="N1079C" part="chapter3" ref="N1079C" type="section">3.3</cms:entry><cms:entry id="_Toc119564339" part="chapter3" ref="_Toc119564339" type="link"/><cms:entry id="N107AB" part="chapter3" ref="N107AB" type="section">3.4</cms:entry><cms:entry id="N107B2" part="chapter3" ref="N107B2" type="citenumber">22</cms:entry><cms:entry id="N107BE" part="chapter3" ref="N107BE" type="citenumber">23</cms:entry><cms:entry id="N107CA" part="chapter3" ref="N107CA" type="citenumber">24</cms:entry><cms:entry id="N107D6" part="chapter3" ref="N107D6" type="citenumber">25</cms:entry><cms:entry id="_Toc119564340" part="chapter3" ref="_Toc119564340" type="link"/><cms:entry id="N107F9" part="chapter3" ref="N107F9" type="section">3.5</cms:entry><cms:entry id="N10805" part="chapter3" ref="N10805" type="citenumber">26</cms:entry><cms:entry id="_Toc119564341" part="chapter3" ref="_Toc119564341" type="link"/><cms:entry id="N1083F" part="chapter3" ref="N1083F" type="section">3.6</cms:entry><cms:entry id="N10846" part="chapter3" ref="N10846" type="citenumber">27</cms:entry><cms:entry id="_Toc119564342" part="chapter3" ref="_Toc119564342" type="link"/><cms:entry id="N10891" part="chapter3" ref="N10891" type="section">3.7</cms:entry><cms:entry id="N10896" part="chapter3" ref="N10896" type="subsection">3.7.1</cms:entry><cms:entry id="_Toc119564343" part="chapter3" ref="_Toc119564343" type="link"/><cms:entry id="N108A0" part="chapter3" ref="N108A0" type="citenumber">28</cms:entry><cms:entry id="_Toc119564344" part="chapter3" ref="_Toc119564344" type="link"/><cms:entry id="N108B3" part="chapter3" ref="N108B3" type="subsection">3.7.2</cms:entry><cms:entry id="N108C0" part="chapter3" ref="N108C0" type="citenumber">29</cms:entry><cms:entry id="_Toc119564345" part="chapter3" ref="_Toc119564345" type="link"/><cms:entry id="N108EA" part="chapter3" ref="N108EA" type="section">3.8</cms:entry><cms:entry id="N108FA" part="chapter3" ref="N108FA" type="citenumber">30</cms:entry><cms:entry id="_Toc119564346" part="chapter3" ref="_Toc119564346" type="link"/><cms:entry id="chapter4" part="chapter4" ref="chapter4" type="chapter">4</cms:entry><cms:entry id="N1090B" part="chapter4" ref="N1090B" type="helpercitenumber">30</cms:entry><cms:entry id="_Toc119564347" part="chapter4" ref="_Toc119564347" type="link"/><cms:entry id="chapter5" part="chapter5" ref="chapter5" type="chapter">5</cms:entry><cms:entry id="N1092A" part="chapter5" ref="N1092A" type="section">5.1</cms:entry><cms:entry id="_Toc119564348" part="chapter5" ref="_Toc119564348" type="link"/><cms:entry id="N10932" part="chapter5" ref="N10932" type="helpercitenumber">30</cms:entry><cms:entry id="N10950" part="chapter5" ref="N10950" type="citenumber">31</cms:entry><cms:entry id="N10953" part="chapter5" ref="N10953" type="table"/><cms:entry id="_Toc119564349" part="chapter5" ref="_Toc119564349" type="link"/><cms:entry id="N10BCE" part="chapter5" ref="N10BCE" type="section">5.2</cms:entry><cms:entry id="N10BD8" part="chapter5" ref="N10BD8" type="citenumber">32</cms:entry><cms:entry id="_Toc119564350" part="chapter5" ref="_Toc119564350" type="link"/><cms:entry id="N10BEA" part="chapter5" ref="N10BEA" type="section">5.3</cms:entry><cms:entry id="N10C17" part="chapter5" ref="N10C17" type="mm">604#299</cms:entry><cms:entry id="N10C22" part="chapter5" ref="N10C22" type="citenumber">33</cms:entry><cms:entry id="N10C6F" part="chapter5" ref="N10C6F" type="mm">615#308</cms:entry><cms:entry id="N10CA0" part="chapter5" ref="N10CA0" type="citenumber">34</cms:entry><cms:entry id="N10CA3" part="chapter5" ref="N10CA3" type="mm">626#286</cms:entry><cms:entry id="N10CB1" part="chapter5" ref="N10CB1" type="mm">614#276</cms:entry><cms:entry id="N10CBC" part="chapter5" ref="N10CBC" type="citenumber">35</cms:entry><cms:entry id="N10CD3" part="chapter5" ref="N10CD3" type="mm">614#307</cms:entry><cms:entry id="N10CEC" part="chapter5" ref="N10CEC" type="citenumber">36</cms:entry><cms:entry id="N10CEF" part="chapter5" ref="N10CEF" type="mm">605#690</cms:entry><cms:entry id="_Toc119564351" part="chapter5" ref="_Toc119564351" type="link"/><cms:entry id="N10D36" part="chapter5" ref="N10D36" type="section">5.4</cms:entry><cms:entry id="N10D40" part="chapter5" ref="N10D40" type="citenumber">37</cms:entry><cms:entry id="N10D43" part="chapter5" ref="N10D43" type="table"/><cms:entry id="_Toc119564352" part="chapter5" ref="_Toc119564352" type="link"/><cms:entry id="chapter6" part="chapter6" ref="chapter6" type="chapter">6</cms:entry><cms:entry id="N1125D" part="chapter6" ref="N1125D" type="helpercitenumber">37</cms:entry><cms:entry id="N11262" part="chapter6" ref="N11262" type="section">6.1</cms:entry><cms:entry id="_Toc119564353" part="chapter6" ref="_Toc119564353" type="link"/><cms:entry id="N1126C" part="chapter6" ref="N1126C" type="citenumber">38</cms:entry><cms:entry id="N1129D" part="chapter6" ref="N1129D" type="citenumber">39</cms:entry><cms:entry id="N112DB" part="chapter6" ref="N112DB" type="citenumber">40</cms:entry><cms:entry id="_Toc119564354" part="chapter6" ref="_Toc119564354" type="link"/><cms:entry id="N112EF" part="chapter6" ref="N112EF" type="section">6.2</cms:entry><cms:entry id="N112F4" part="chapter6" ref="N112F4" type="subsection">6.2.1</cms:entry><cms:entry id="_Toc119564355" part="chapter6" ref="_Toc119564355" type="link"/><cms:entry id="N1130A" part="chapter6" ref="N1130A" type="citenumber">41</cms:entry><cms:entry id="N1134D" part="chapter6" ref="N1134D" type="citenumber">42</cms:entry><cms:entry id="_Toc119564356" part="chapter6" ref="_Toc119564356" type="link"/><cms:entry id="N1138C" part="chapter6" ref="N1138C" type="subsection">6.2.2</cms:entry><cms:entry id="N113C8" part="chapter6" ref="N113C8" type="citenumber">43</cms:entry><cms:entry id="_Toc119564357" part="chapter6" ref="_Toc119564357" type="link"/><cms:entry id="N11419" part="chapter6" ref="N11419" type="subsection">6.2.3</cms:entry><cms:entry id="_Toc119564358" part="chapter6" ref="_Toc119564358" type="link"/><cms:entry id="N1149C" part="chapter6" ref="N1149C" type="subsection">6.2.4</cms:entry><cms:entry id="N114A6" part="chapter6" ref="N114A6" type="citenumber">44</cms:entry><cms:entry id="_Toc119564359" part="chapter6" ref="_Toc119564359" type="link"/><cms:entry id="N114D8" part="chapter6" ref="N114D8" type="subsection">6.2.5</cms:entry><cms:entry id="_Toc119564360" part="chapter6" ref="_Toc119564360" type="link"/><cms:entry id="N11529" part="chapter6" ref="N11529" type="subsection">6.2.6</cms:entry><cms:entry id="N11576" part="chapter6" ref="N11576" type="citenumber">45</cms:entry><cms:entry id="_Toc119564361" part="chapter6" ref="_Toc119564361" type="link"/><cms:entry id="N115A3" part="chapter6" ref="N115A3" type="subsection">6.2.7</cms:entry><cms:entry id="_Toc119564362" part="chapter6" ref="_Toc119564362" type="link"/><cms:entry id="N115CB" part="chapter6" ref="N115CB" type="subsection">6.2.8</cms:entry><cms:entry id="_Toc119564363" part="chapter6" ref="_Toc119564363" type="link"/><cms:entry id="N115F9" part="chapter6" ref="N115F9" type="subsection">6.2.9</cms:entry><cms:entry id="N11603" part="chapter6" ref="N11603" type="citenumber">46</cms:entry><cms:entry id="_Toc119564364" part="chapter6" ref="_Toc119564364" type="link"/><cms:entry id="N1162C" part="chapter6" ref="N1162C" type="subsection">6.2.10</cms:entry><cms:entry id="_Toc119564365" part="chapter6" ref="_Toc119564365" type="link"/><cms:entry id="N1164D" part="chapter6" ref="N1164D" type="subsection">6.2.11</cms:entry><cms:entry id="N11659" part="chapter6" ref="N11659" type="citenumber">47</cms:entry><cms:entry id="N116E2" part="chapter6" ref="N116E2" type="citenumber">48</cms:entry><cms:entry id="_Toc119564366" part="chapter6" ref="_Toc119564366" type="link"/><cms:entry id="N11719" part="chapter6" ref="N11719" type="section">6.3</cms:entry><cms:entry id="_Toc119564367" part="chapter6" ref="_Toc119564367" type="link"/><cms:entry id="N1175A" part="chapter6" ref="N1175A" type="section">6.4</cms:entry><cms:entry id="N1176A" part="chapter6" ref="N1176A" type="citenumber">49</cms:entry><cms:entry id="N1177E" part="chapter6" ref="N1177E" type="citenumber">50</cms:entry><cms:entry id="_Toc119564368" part="chapter6" ref="_Toc119564368" type="link"/><cms:entry id="chapter7" part="chapter7" ref="chapter7" type="chapter">7</cms:entry><cms:entry id="N1178F" part="chapter7" ref="N1178F" type="helpercitenumber">50</cms:entry><cms:entry id="_Toc119564369" part="chapter7" ref="_Toc119564369" type="link"/><cms:entry id="chapter8" part="chapter8" ref="chapter8" type="chapter">8</cms:entry><cms:entry id="N117F7" part="chapter8" ref="N117F7" type="helpercitenumber">50</cms:entry><cms:entry id="_Toc119564370" part="chapter8" ref="_Toc119564370" type="link"/><cms:entry id="chapter9" part="chapter9" ref="chapter9" type="chapter">9</cms:entry><cms:entry id="N11851" part="chapter9" ref="N11851" type="section">9.1</cms:entry><cms:entry id="_Toc119564371" part="chapter9" ref="_Toc119564371" type="link"/><cms:entry id="N1185B" part="chapter9" ref="N1185B" type="citenumber">51</cms:entry><cms:entry id="N1185E" part="chapter9" ref="N1185E" type="table"/><cms:entry id="N11F69" part="chapter9" ref="N11F69" type="table"/><cms:entry id="_Toc119564372" part="chapter9" ref="_Toc119564372" type="link"/><cms:entry id="N1265A" part="chapter9" ref="N1265A" type="section">9.2</cms:entry><cms:entry id="N1265F" part="chapter9" ref="N1265F" type="subsection">9.2.1</cms:entry><cms:entry id="_Toc119564373" part="chapter9" ref="_Toc119564373" type="link"/><cms:entry id="N12667" part="chapter9" ref="N12667" type="block">9.2.1.1</cms:entry><cms:entry id="_Toc119564374" part="chapter9" ref="_Toc119564374" type="link"/><cms:entry id="N12674" part="chapter9" ref="N12674" type="citenumber">52</cms:entry><cms:entry id="N1267D" part="chapter9" ref="N1267D" type="table"/><cms:entry id="N127AB" part="chapter9" ref="N127AB" type="table"/><cms:entry id="N12851" part="chapter9" ref="N12851" type="citenumber">53</cms:entry><cms:entry id="N12857" part="chapter9" ref="N12857" type="table"/><cms:entry id="N128C8" part="chapter9" ref="N128C8" type="citenumber">54</cms:entry><cms:entry id="N128CB" part="chapter9" ref="N128CB" type="table"/><cms:entry id="_Toc119564375" part="chapter9" ref="_Toc119564375" type="link"/><cms:entry id="N12934" part="chapter9" ref="N12934" type="block">9.2.1.2</cms:entry><cms:entry id="N1293B" part="chapter9" ref="N1293B" type="table"/><cms:entry id="_Toc119564376" part="chapter9" ref="_Toc119564376" type="link"/><cms:entry id="N129B0" part="chapter9" ref="N129B0" type="block">9.2.1.3</cms:entry><cms:entry id="N129B7" part="chapter9" ref="N129B7" type="table"/><cms:entry id="_Toc119564377" part="chapter9" ref="_Toc119564377" type="link"/><cms:entry id="N12A56" part="chapter9" ref="N12A56" type="block">9.2.1.4</cms:entry><cms:entry id="N12A5D" part="chapter9" ref="N12A5D" type="citenumber">55</cms:entry><cms:entry id="N12A63" part="chapter9" ref="N12A63" type="table"/><cms:entry id="N12AB1" part="chapter9" ref="N12AB1" type="citenumber">56</cms:entry><cms:entry id="N12AB4" part="chapter9" ref="N12AB4" type="table"/><cms:entry id="_Toc119564378" part="chapter9" ref="_Toc119564378" type="link"/><cms:entry id="N12B26" part="chapter9" ref="N12B26" type="block">9.2.1.5</cms:entry><cms:entry id="N12B2D" part="chapter9" ref="N12B2D" type="table"/><cms:entry id="_Toc119564379" part="chapter9" ref="_Toc119564379" type="link"/><cms:entry id="N12B6F" part="chapter9" ref="N12B6F" type="block">9.2.1.6</cms:entry><cms:entry id="N12B7C" part="chapter9" ref="N12B7C" type="citenumber">57</cms:entry><cms:entry id="N12B7F" part="chapter9" ref="N12B7F" type="table"/><cms:entry id="N12BB2" part="chapter9" ref="N12BB2" type="table"/><cms:entry id="N12BF1" part="chapter9" ref="N12BF1" type="citenumber">58</cms:entry><cms:entry id="N12BF4" part="chapter9" ref="N12BF4" type="table"/><cms:entry id="N12C8F" part="chapter9" ref="N12C8F" type="table"/><cms:entry id="_Toc119564380" part="chapter9" ref="_Toc119564380" type="link"/><cms:entry id="N12CDF" part="chapter9" ref="N12CDF" type="subsection">9.2.2</cms:entry><cms:entry id="N12CE4" part="chapter9" ref="N12CE4" type="block">9.2.2.1</cms:entry><cms:entry id="_Toc119564381" part="chapter9" ref="_Toc119564381" type="link"/><cms:entry id="N12CEE" part="chapter9" ref="N12CEE" type="citenumber">59</cms:entry><cms:entry id="_Toc119564382" part="chapter9" ref="_Toc119564382" type="link"/><cms:entry id="N12CFD" part="chapter9" ref="N12CFD" type="block">9.2.2.2</cms:entry><cms:entry id="_Toc119564383" part="chapter9" ref="_Toc119564383" type="link"/><cms:entry id="N12D10" part="chapter9" ref="N12D10" type="block">9.2.2.3</cms:entry><cms:entry id="_Toc119564384" part="chapter9" ref="_Toc119564384" type="link"/><cms:entry id="N12D23" part="chapter9" ref="N12D23" type="block">9.2.2.4</cms:entry><cms:entry id="N12D2A" part="chapter9" ref="N12D2A" type="citenumber">60</cms:entry><cms:entry id="_Toc119564385" part="chapter9" ref="_Toc119564385" type="link"/><cms:entry id="N12D3B" part="chapter9" ref="N12D3B" type="block">9.2.2.5</cms:entry><cms:entry id="_Toc119564386" part="chapter9" ref="_Toc119564386" type="link"/><cms:entry id="N12D4C" part="chapter9" ref="N12D4C" type="block">9.2.2.6</cms:entry><cms:entry id="_Toc119564387" part="chapter9" ref="_Toc119564387" type="link"/><cms:entry id="N12D5D" part="chapter9" ref="N12D5D" type="block">9.2.2.7</cms:entry><cms:entry id="N12D64" part="chapter9" ref="N12D64" type="citenumber">61</cms:entry><cms:entry id="_Toc119564388" part="chapter9" ref="_Toc119564388" type="link"/><cms:entry id="N12D71" part="chapter9" ref="N12D71" type="block">9.2.2.8</cms:entry><cms:entry id="_Toc119564389" part="chapter9" ref="_Toc119564389" type="link"/><cms:entry id="N12D82" part="chapter9" ref="N12D82" type="block">9.2.2.9</cms:entry><cms:entry id="_Toc119564390" part="chapter9" ref="_Toc119564390" type="link"/><cms:entry id="N12D99" part="chapter9" ref="N12D99" type="block">9.2.2.10</cms:entry><cms:entry id="N12DA0" part="chapter9" ref="N12DA0" type="citenumber">62</cms:entry><cms:entry id="_Toc119564391" part="chapter9" ref="_Toc119564391" type="link"/><cms:entry id="N12DB1" part="chapter9" ref="N12DB1" type="block">9.2.2.11</cms:entry><cms:entry id="_Toc119564392" part="chapter9" ref="_Toc119564392" type="link"/><cms:entry id="N12DC9" part="chapter9" ref="N12DC9" type="subsection">9.2.3</cms:entry><cms:entry id="N12DCE" part="chapter9" ref="N12DCE" type="block">9.2.3.1</cms:entry><cms:entry id="_Toc119564393" part="chapter9" ref="_Toc119564393" type="link"/><cms:entry id="_Toc119564394" part="chapter9" ref="_Toc119564394" type="link"/><cms:entry id="N12DE4" part="chapter9" ref="N12DE4" type="block">9.2.3.2</cms:entry><cms:entry id="N12DEB" part="chapter9" ref="N12DEB" type="citenumber">63</cms:entry><cms:entry id="_Toc119564395" part="chapter9" ref="_Toc119564395" type="link"/><cms:entry ref="N12E06" type="back"/><cms:entry id="N12E08" part="N12E08" ref="N12E08" type="bibliography">References</cms:entry><cms:entry id="N1515E" part="N1515E" ref="N1515E" type="acknowledgement">
            
            Danksagung</cms:entry><cms:entry id="_Toc471900148" part="N1515E" ref="_Toc471900148" type="link"/><cms:entry id="_Toc119564396" part="N1515E" ref="_Toc119564396" type="link"/><cms:entry id="N15176" part="N15176" ref="N15176" type="vita">
            
            Lebenslauf</cms:entry><cms:entry id="_Toc471900149" part="N15176" ref="_Toc471900149" type="link"/><cms:entry id="_Toc119564397" part="N15176" ref="_Toc119564397" type="link"/><cms:entry id="N15183" part="N15176" ref="N15183" type="table"/><cms:entry id="N1529A" part="N1529A" ref="N1529A" type="declaration">
            
            Eidestattliche Erklärung</cms:entry><cms:entry id="_Toc471900150" part="N1529A" ref="_Toc471900150" type="link"/><cms:entry id="_Toc119564398" part="N1529A" ref="_Toc119564398" type="link"/><cms:entry part="front" type=":current"/><cms:entry type=":lang">en</cms:entry><cms:entry ref=":contents" type=":contents">Table of contents</cms:entry><cms:entry type=":help"><url href="http://...">Help</url></cms:entry></cms:meta><cms:content><front id="front"><school>Aus dem Institut für Mikrobiologie und Hygiene<br/>der Medizinischen Fakultät der Charité - Universitätsmedizin Berlin<br/>und<br/>aus der Abteilung für Parodontologie und Synoptische Zahnmedizin<br/>Zentrum für Zahnmedizin der Medizinischen Fakultät der Charité -<br/>Universitätsmedizin Berlin</school><p>
         <link id="_Toc119564310"/>
      </p><submission>DISSERTATION</submission><degree>zur Erlangung des akademischen Grades<br/>Doctor medicinae dentariae<br/>(Dr. med. dent.)</degree><title>Prevalence analysis of putative periodontal pathogens<br/>in patients with aggressive periodontitis and healthy elderly.<br/>A molecular study</title><major>vorgelegt der Medizinischen Fakultät der Charité -<br/>Universitätsmedizin Berlin</major><author>von Zahnärztin <given>Lilian </given>
         <surname>Edesi-Neuss</surname><br/><suffix>aus Tallinn, Estland</suffix>
      </author><dean>Dekan: Prof. Dr. med. Martin Paul</dean><approvals>
         <name>Prof. Dr. Dr. U. B. Göbel</name>
         <name>Prof. Dr. Dr. J.-P. Bernimoulin</name>
         <name>Prof. Dr. G. Krekeler </name>
      </approvals><date>Datum der Promotion:21.11.05</date><abstract lang="de">
         <head>
            <link id="_Toc119564311"/>Exzerpt</head>
         <p>Marginale Parodontitis, die multikausale Erkrankung des Parodonts ist in erster Linie eine Infektionskrankheit, modifiziert durch Wirtsfaktoren und äußere Einflüße. Die als pathogene Mischflora bezeichnete Kombination kommensaler Mikroorganismen, die opportunistische Infektionen und damit Immunreaktionen auslösen können, spielt die primäre Rolle in der Ätiopathogenese der Parodontitis. In der Aufstellung des Studienziels wurden einzelne, vermutlich pathogene Bakterienarten (<em>Tannerella forsythensis</em>, <em>Porphyromonas gingivalis</em>, <em>Actinobacillus actinomycetemcomitans</em>, <em>Campylobacter rectus</em>, <em>Fusobacterium nucleatum</em>, <em>Fusobacterium</em> spp., <em>Prevotella intermedia</em>, <em>Eikenella corrodens</em>, <em>Veillonella parvula</em> und <em>Capnocytophaga ochracea</em>) auf Grund der Evidenz von früheren Studien ausgewählt, die eventuell als "Markerkeime" in der aggressiven Form der Parodontitis betrachtet werden können. Dazu wurde eine Kontrollgruppe (die Senioren) untersucht, die eine gesunde parodontale Flora besitzen. Die angewandte Nachweismethode basiert auf eubakterieller PCR-Amplifikation von 16S rDNA und darauffolgender dot-blot Hybridisierung mit spezifischen Oligonukleotidsonden. Die entsprechenden Sonden wurden hergestellt und evaluiert. Die Optimierung der Hybridisierungsbedingungen einzelner Sonden folgte unter Einsatz von Positiv- und Negativkontrollen - PCR-Produkte von 42 gezüchteten Ziel- bzw. phylogenetisch nahliegenden Baktetrienstämmen. Für die epidemiologische Untersuchung wurde subgingivale Plaque von vier Parodontaltaschen und einer Kontrollstelle von 45 Patienten mit aggressiver Parodontitis, sowie an fünf Stellen von 21 Senioren entnommen. Die Prävalenz der einzelnen Bakterienarten wurde mit Hilfe des Chi-quadrat-Tests verglichen. Die Ergebnisse dieser Studie bewiesen die erfolgreiche Einsetzbarkeit der hergestellten Oligonukleotidsonden. Es konnte gezeigt werden, daß alle untersuchten Bakterien in beiden Gruppen vorkommen. Obgleich eine hohe interindividuelle Variabilität der Kolonisationsmuster zu beobachten war, konnten <em>T. forsythensis</em>, <em>P. gingivalis</em> und <em>F. nucleatum</em> sehr häufig in den Parodontaltaschen nachgewiesen werden. Obwohl diese Arten auch an den gesunden Stellen der parodontal Erkrankten sowie der Senioren festzustellen waren,  blieb die Häufigkeit  dieser Besiedlung signifikant seltener. </p>
         <p>
            <em>A. actinomycetemcomitans</em>  konnte nur bei einzelnen Patienten mit aggressiver Parodontitis festgestellt werden. Die Ergebnisse für <em>P. intermedia</em> und <em>E. corrodens</em> ließen keine eindeutige Assoziation sowohl mit der aggressiven Parodontitis als auch mit dem gesunden Parodontalzustand zu. Bei Senioren wurde <em>C. ochracea</em> besonders häufig nachgewiesen. Zusammenfassend kann man die vermutlichen Parodontalpathogene wie <em>T. forsythensis</em>, <em>P. gingivalis</em>, <em>F. nucleatum</em> und <em>C. rectus</em> als Leitkeime aggressiver Parodontitis ansehen. Bezüglich der polymikrobiellen Natur der Parodontitis würde eine umfassende Untersuchung der oralen Mikroflora und deren Zusammenspiel mit den Wirtsfaktoren zur Aufklärung der Ätiopathogenese der Parodontitis eher beitragen als der Nachweis einzelner Arten.</p>
         <p>Schlagwörter: </p>
         <p>
            <link id="_Toc119564312"/>
         </p>
      </abstract><keywords lang="de">
         <keyword>Parodontalpathogene</keyword>
         <keyword>aggressive Parodontitis</keyword>
         <keyword>Oligonukleotidsonden</keyword>
         <keyword>Dot-blot Hybridisierung</keyword>
         <keyword>PCR-Amplifikation</keyword>
      </keywords><abstract lang="en">
         <head>Abstract</head>
         <p>A multifactorial risk pattern of periodontitis has been recognized, where in addition to host and environmental factors a pathogenic microbiota plays a primary role. At present no definite answer can be given to the question of whether the expression of either aggressive etiological agents (implying infection with a virulent microbiota), or a high level of individual susceptibility to periodontal disease, or a specific combination of both is the conductive factor in the etiopathogenesis of aggressive periodontitis. The purpose of the current research was to analyze the prevalence of periodontitis-associated microorganisms in patients with aggressive periodontitis and periodontally healthy elders by using molecular-biologic detection methods like eubacterial PCR-amplification of 16S rDNA in combination with dot-blot hybridization. The oligonucleotide probes for the detection of <em>Tannerella forsythensis</em>, <em>Porphyromonas gingivalis</em>, <em>Actinobacillus actinomycetemcomitans</em>, <em>Campylobacter rectus</em>, <em>Fusobacterium nucleatum</em>, <em>Fusobacterium</em> spp., <em>Prevotella intermedia</em>, <em>Eikenella corrodens</em>, <em>Veillonella parvula</em> and <em>Capnocytophaga ochracea</em> were designed and evaluated. The PCR products of 42 cultivated target and closely related bacteria were obtained for the optimization of hybridization conditions. For the epidemiological study subgingival plaque was sampled from four pockets and one healthy site of 45 aggressive periodontitis patients as well as from five sites of 21 elderly. The differences in the prevalence of bacterial species was analyzed by chi-square test. The results of the study confirmed the reliability of the oligonucleotide probes in a specific and sensitive detection of the respective oral species. The data of the epidemiological study revealed frequent colonization by <em>T. forsythensis</em>, <em>P. gingivalis</em>, <em>F. nucleatum</em> and <em>C. rectus</em> in patients with aggressive periodontitis, however individual variations were obvious. These microorganisms could be predominantly identified in periodontal pockets, but were significantly less common in the healthy sites of the periodontitis patients and in the elderly subjects. <em>A. actinomycetemcomitans</em> could be detected in only a few patients, reducing its suspected importance in the etiopathogenesis of aggressive periodontitis. No direct association for <em>P. intermedia</em> and <em>E. corrodens</em> with aggressive periodontitis or periodontal health could be seen. <em>C. ochracea</em> was highly prevalent in the well-maintained elderly, being rarely found in the diseased group. The putative pathogens <em>T. forsythensis</em>, <em>P. gingivalis</em>, <em>F. nucleatum</em> and <em>C. rectus</em> can be conclusively suggested as the key-bacteria in patients with aggressive periodontitis. However, considering that periodontitis is a polymicrobial infection, the screening of the microbial population, rather than the isolation of single members of the subgingival flora, should give a more comprehensive perspective in etiopathogenetic research of periodontitis.</p>
         <p>Keywords: </p>
      </abstract><keywords lang="en">
         <keyword>periodontal pathogens</keyword>
         <keyword>16S rRNA</keyword>
         <keyword>oligonucleotide probes</keyword>
         <keyword>dot-blot hybridization</keyword>
         <keyword>aggressive periodontitis</keyword>
         <keyword>PCR-amplification</keyword>
      </keywords><freehead id=":contents">Table of contents</freehead><ul><li><p><link ref="chapter1">1</link> 
            Introduction<ul><li><p><link ref="N10135">1.1</link> 
               Classification of periodontal diseases</p></li><li><p><link ref="N101A3">1.2</link> Bacterial etiology of periodontal diseases<ul><li><p><link ref="N1020D">1.2.1</link> 
                  Healthy flora</p></li><li><p><link ref="N1021F">1.2.2</link> Gingivitis </p></li><li><p><link ref="N10237">1.2.3</link> Periodontitis</p></li><li><p><link ref="N103A2">1.2.4</link> Bacterial consortia</p></li><li><p><link ref="N103EE">1.2.5</link> Biofilm</p></li><li><p><link ref="N103FF">1.2.6</link> Virulence factors</p></li></ul></p></li><li><p><link ref="N104A0">1.3</link> Pathogenesis of periodontitis</p></li><li><p><link ref="N104D1">1.4</link> Clinical studies seeking evidence for etiological role of bacteria<ul><li><p><link ref="N104DE">1.4.1</link> 
                  Prevalence studies</p></li><li><p><link ref="N10502">1.4.2</link> Progression of disease</p></li><li><p><link ref="N1053F">1.4.3</link> Risk factor studies</p></li><li><p><link ref="N1055A">1.4.4</link> Treatment studies</p></li></ul></p></li><li><p><link ref="N10586">1.5</link> Detection methods</p></li><li><p><link ref="N1059D">1.6</link> Aggressive periodontitis<ul><li><p><link ref="N105A2">1.6.1</link> 
                  Clinical diagnosis</p></li><li><p><link ref="N10646">1.6.2</link> Microorganisms associated with aggressive periodontitis</p></li></ul></p></li><li><p><link ref="N10677">1.7</link> Control group (Elderly)</p></li></ul></p></li><li><p><link ref="chapter2">2</link> 
            Aim of the study</p></li><li><p><link ref="chapter3">3</link> 
            Materials and methods<ul><li><p><link ref="N106C2">3.1</link> 
               Probe design</p></li><li><p><link ref="N106FA">3.2</link> Culturing of target- and phylogenetically closely related species</p></li><li><p><link ref="N1079C">3.3</link> DNA isolation</p></li><li><p><link ref="N107AB">3.4</link> PCR amplification of 16S rDNA</p></li><li><p><link ref="N107F9">3.5</link> Probe labeling</p></li><li><p><link ref="N1083F">3.6</link> Optimization of hybridization conditions</p></li><li><p><link ref="N10891">3.7</link> Patient populations<ul><li><p><link ref="N10896">3.7.1</link> 
                  GAP patients</p></li><li><p><link ref="N108B3">3.7.2</link> Elderly subjects</p></li></ul></p></li><li><p><link ref="N108EA">3.8</link> Dot-blot hybridization of patient material</p></li></ul></p></li><li><p><link ref="chapter4">4</link> Statistical analysis</p></li><li><p><link ref="chapter5">5</link> Results<ul><li><p><link ref="N1092A">5.1</link> 
               Design of species-specific oligonucleotide probes</p></li><li><p><link ref="N10BCE">5.2</link> Optimization of the hybridization conditions and evaluation of the oligonucleotide probes</p></li><li><p><link ref="N10BEA">5.3</link> Epidemiological study - GAP patients and healthy elderly individuals</p></li><li><p><link ref="N10D36">5.4</link> Bacterial consortia</p></li></ul></p></li><li><p><link ref="chapter6">6</link> Discussion<ul><li><p><link ref="N11262">6.1</link> 
               Detection methods</p></li><li><p><link ref="N112EF">6.2</link> Epidemiological study<ul><li><p><link ref="N112F4">6.2.1</link> 
                  
                  <em>Tannerella forsythensis</em>
               </p></li><li><p><link ref="N1138C">6.2.2</link> 
                  <em>Porphyromonas gingivalis</em>
               </p></li><li><p><link ref="N11419">6.2.3</link> 
                  <em>Actinobacillus actinomycetemcomitans</em>
               </p></li><li><p><link ref="N1149C">6.2.4</link> 
                  <em>Campylobacter rectus</em>
               </p></li><li><p><link ref="N114D8">6.2.5</link> 
                  <em>Fusobacterium nucleatum</em>
               </p></li><li><p><link ref="N11529">6.2.6</link> 
                  <em>Prevotella intermedia</em>
               </p></li><li><p><link ref="N115A3">6.2.7</link> 
                  <em>Eikenella corrodens</em>
               </p></li><li><p><link ref="N115CB">6.2.8</link> 
                  <em>Veillonella parvula</em>
               </p></li><li><p><link ref="N115F9">6.2.9</link> 
                  <em>Capnocytophaga ochracea</em>
               </p></li><li><p><link ref="N1162C">6.2.10</link> Colonization of healthy sites</p></li><li><p><link ref="N1164D">6.2.11</link> Bacterial consortia</p></li></ul></p></li><li><p><link ref="N11719">6.3</link> Additional species associated with periodontitis</p></li><li><p><link ref="N1175A">6.4</link> Host factors</p></li></ul></p></li><li><p><link ref="chapter7">7</link> Summary</p></li><li><p><link ref="chapter8">8</link> Zusammenfassung</p></li><li><p><link ref="chapter9">9</link> Supplement<ul><li><p><link ref="N11851">9.1</link> 
               Tables</p></li><li><p><link ref="N1265A">9.2</link> List of used mediums and chemicals<ul><li><p><link ref="N1265F">9.2.1</link> 
                  Mediums<ul><li><p><link ref="N12667">9.2.1.1</link> 
                     FUM - Fluid Universal Medium</p></li><li><p><link ref="N12934">9.2.1.2</link> Columbia agar</p></li><li><p><link ref="N129B0">9.2.1.3</link> TSBV - Trypticase-Soy-Bacitracin-Agar (J. Slots, J Clin Microbiol 1982: 15: 606)</p></li><li><p><link ref="N12A56">9.2.1.4</link> ETSA - Enriched-Trypticase-Soy-Agar</p></li><li><p><link ref="N12B26">9.2.1.5</link> Trypticase Soy Yeast Extract Medium</p></li><li><p><link ref="N12B6F">9.2.1.6</link> RTF</p></li></ul></p></li><li><p><link ref="N12CDF">9.2.2</link> Buffers<ul><li><p><link ref="N12CE4">9.2.2.1</link> 
                     Maleic acid buffer</p></li><li><p><link ref="N12CFD">9.2.2.2</link> 1%  Blocking solution</p></li><li><p><link ref="N12D10">9.2.2.3</link> Detection buffer</p></li><li><p><link ref="N12D23">9.2.2.4</link> Hybridization buffer</p></li><li><p><link ref="N12D3B">9.2.2.5</link> Washing buffer 0:</p></li><li><p><link ref="N12D4C">9.2.2.6</link> Washing buffer 1:</p></li><li><p><link ref="N12D5D">9.2.2.7</link> Washing buffer 2:</p></li><li><p><link ref="N12D71">9.2.2.8</link> Stripping solution</p></li><li><p><link ref="N12D82">9.2.2.9</link> Phosphate buffered saline (PBS)</p></li><li><p><link ref="N12D99">9.2.2.10</link> Lysis buffer</p></li><li><p><link ref="N12DB1">9.2.2.11</link> Stop-Solution</p></li></ul></p></li><li><p><link ref="N12DC9">9.2.3</link> Gels<ul><li><p><link ref="N12DCE">9.2.3.1</link> 
                     1.2% Agarose gel</p></li><li><p><link ref="N12DE4">9.2.3.2</link> Polyacrylamidegel</p></li></ul></p></li></ul></p></li></ul></p></li><li><p><link ref="N12E08">References</link></p></li><li><p><link ref="N1515E">
            
            Danksagung</link></p></li><li><p><link ref="N15176">
            
            Lebenslauf</link></p></li><li><p><link ref="N1529A">
            
            Eidestattliche Erklärung</link></p></li></ul><freehead id=":toc-tables">Tables</freehead><ul><li><p><link ref="N10248">Table 1. Bacterial species associated with chronic and aggressive periodontitis.</link></p></li><li><p><link ref="N10953">Table 1. Probe sequences (5'-3') with respective hybridization conditions. Washing buffers listed in Suppl., 2.5.-2.7.</link></p></li><li><p><link ref="N10D43">Table. 2. Odds ratios of associations among investigated species. The value 0 is the result of the absence of one species in any site in one group.</link></p></li><li><p><link ref="N1185E">Table 1. Strain designations, original sources, control identification method used in the study.</link></p></li><li><p><link ref="N11F69">Table 2. The bacterial species, respective culture media and growth conditions.</link></p></li></ul><freehead id=":toc-media">Images</freehead><ul><li><p><link ref="N10C17">Fig. 1 Bar chart showing the prevalence of the respective species in 44 GAP patients (one GAP patient was excluded because of a missing control site) and 21 elderly subjects. A patient was regarded positive when at least one site harbored the respective species. The significance of differences between the groups was calculated using the chi-square test. </link></p></li><li><p><link ref="N10C6F">Fig. 2. Percent of positive periodontal pockets and control sites in 44 GAP patients. The significance of differences was evaluated using the chi-square test. </link></p></li><li><p><link ref="N10CA3">Fig. 3. Site-prevalence of the species in 45 GAP patients and 21 elderly (4 sites per subject were included). The significance of the differences between the groups was determined using the chi-square test. </link></p></li><li><p><link ref="N10CB1">Fig. 4. Comparison between the colonization of shallow sites (PD 1-3 mm) in 44 GAP patients and 17 elderly. Only one site per subject could be included. The significance of differences was evaluated using the chi-square test. </link></p></li><li><p><link ref="N10CD3">Fig. 5. Bar chart of the percent of positive sites with probing depth 1-3 mm, 4-7 mm and 8-12 mm in GAP patients (n=23). The significance of differences was determined using the chi-square test. </link></p></li><li><p><link ref="N10CEF">Fig.6. The bars depict the percent of subjects with either 0, 1, 2, 3 or 4 positive sites for <em>T. forsythensis</em>, <em>P. gingivalis</em>, <em>F. nucleatum</em>, <em>P. intermedia</em>, <em>E. corrodens</em> and <em>C. ochracea</em> in GAP group (n=45) and elderly (n=21). The significance of differences between the groups were determined using the Mann-Whitney test. </link></p></li></ul></front></cms:content></cms:document></cms:container>