Group UIBO (Oral Basic Research Unit), Faculty of Odontology, University of El Bosque, Bogota, Colombia.
J Clin Periodontol. 2011 May;38(5):418-27. doi: 10.1111/j.1600-051X.2011.01717.x. Epub 2011 Mar 11.
The aim of this study was to assess the presence of subgingival pathogens in peripheral blood samples from periodontitis patients before and after scaling and root planing (Sc/RP) using nested polymerase chain reaction (nested PCR).
Peripheral blood samples were obtained from 42 patients with severe generalized chronic or aggressive periodontitis. In each patient, four samples of peripheral blood were drawn at different times: immediately before the Sc/RP procedure; immediately after Sc/RP; 15 and 30 min. post-Sc/RP. Blood samples were analysed for bacteraemia with anaerobic culturing and nested PCR, using universal bacterial primers that target the 16S-rRNA gene of most bacteria, subsequently re-amplified with specific primers to Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, Tannerella forsythia, Eikenella corrodens, Campylobacter rectus and Prevotella intermedia, using a modified phenol-chloroform method for DNA extraction.
Presence of specific periodontal pathogens in peripheral blood after treatment was detected in 54.8% of the patients, in 47.6% with anaerobic culturing and in 19% with nested PCR. In 16.6%, the periodontal pathogens were detected before Sc/RP. P. gingivalis and A. actynomicetemcomitans were the pathogens most frequently detected in the bloodstream before and after Sc/RP.
Nested PCR demonstrated the presence of DNA from periodontal pathogens in blood samples in severe periodontitis patients before, during and after periodontal therapy. The use of these molecular-based techniques may improve the accuracy from the results obtained by haemoculture.
本研究旨在使用巢式聚合酶链反应(nested PCR)评估牙周炎患者在牙周刮治和根面平整(Sc/RP)前后龈下病原体在外周血样本中的存在。
从 42 名患有严重广泛性慢性或侵袭性牙周炎的患者中获得外周血样本。在每个患者中,在不同时间从外周血中抽取了四个样本:在 Sc/RP 程序之前立即;Sc/RP 后立即;Sc/RP 后 15 和 30 分钟。使用针对大多数细菌 16S-rRNA 基因的通用细菌引物对血液样本进行需氧培养和巢式 PCR 分析,随后使用特异性引物对牙龈卟啉单胞菌、伴放线放线杆菌、福赛坦纳菌、侵蚀艾肯菌、直肠弯曲杆菌和中间普氏菌进行重新扩增,使用改良的酚-氯仿法进行 DNA 提取。
在治疗后,54.8%的患者外周血中检测到特定的牙周病原体,厌氧培养的检出率为 47.6%,巢式 PCR 的检出率为 19%。在 16.6%的患者中,牙周病原体在 Sc/RP 之前被检测到。牙龈卟啉单胞菌和伴放线放线杆菌是 Sc/RP 前后最常在外周血中检测到的病原体。
巢式 PCR 显示在牙周炎治疗前后严重牙周炎患者的血液样本中存在牙周病原体的 DNA。使用这些基于分子的技术可以提高血培养结果的准确性。