Comparison of the detection of periodontal pathogens in bacteraemia after tooth brushing by culture and molecular techniques
Comparison of the detection of periodontal pathogens in bacteraemia after tooth brushing by culture and molecular techniques
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DOI：
10.4317/medoral.20842
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发表时间：
2016-05-01
影响因子：
2.2
通讯作者：
Sanz, Mariano
中科院分区：
文献类型：
--
作者：
Marin, Maria-Jose;Figuero, Elena;Sanz, Mariano
背景：本研究比较了4种诊断技术(3种基于培养和1种基于分子技术)诱导刷牙后菌血症标本中牙周病原菌的患病率和数量。材料和方法：采集36例不同牙周状况的受试者(17例健康牙周，10例牙周炎，9例牙周炎)在刷牙前和刷牙后2分钟采集血样。每个样本用三种培养方法[直接厌氧培养(DAC)、血液培养(BACTEC)和裂解-离心法(LC)]和一种基于分子的技术[定量聚合酶链式反应(QPCR)]进行分析。用培养法可以检测和定量任何细菌分离株，而用qPCR只能检测和定量牙龈卟啉单胞菌和伴生放线杆菌。结果：BACTEC和qPCR均未在血样中检出细菌。仅有LC(2.7%)和DAC(8.3%)检出菌血症，尽管不在同一患者中。结论：用4种不同的微生物检测方法对同一样本进行检测结果不同，说明需要对菌血症样本中牙周病原体的检测方法进行适当的验证，主要是在刷牙后血液样本中牙周病原体很少出现的情况下。
Background: The prevalence and amounts of periodontal pathogens detected in bacteraemia samples after tooth brushing-induced by means of four diagnostic technique, three based on culture and one in a molecular-based technique, have been compared in this study.Material and Methods: Blood samples were collected from thirty-six subjects with different periodontal status (17 were healthy, 10 with gingivitis and 9 with periodontitis) at baseline and 2 minutes after tooth brushing. Each sample was analyzed by three culture-based methods [direct anaerobic culturing (DAC), hemo-culture (BACTEC), and lysis-centrifugation (LC)] and one molecular-based technique [quantitative polymerase chain reaction (qPCR)]. With culture any bacterial isolate was detected and quantified, while with qPCR only Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans were detected and quantified. Descriptive analyses, ANOVA and Chi-squared tests, were performed.Results: Neither BACTEC nor qPCR detected any type of bacteria in the blood samples. Only LC (2.7%) and DAC (8.3%) detected bacteraemia, although not in the same patients. Fusobacterium nucleatum was the most frequently detected bacterial species.Conclusions: The disparity in the results when the same samples were analyzed with four different microbiological detection methods highlights the need for a proper validation of the methodology to detect periodontal pathogens in bacteraemia samples, mainly when the presence of periodontal pathogens in blood samples after tooth brushing was very seldom.