Quantitative Real-Time PCR for Porphyromonas gingivalis and Total Bacteria SHARON
Quantitative Real-Time PCR for Porphyromonas gingivalis and Total Bacteria SHARON
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牙龈卟啉单胞菌和总菌 SHARON 的实时定量 PCR
DOI：
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发表时间：
2000
期刊：
影响因子：
--
通讯作者：
J. Leys
中科院分区：
文献类型：
--
作者：
R. Lyons;A. Griffen;J. Leys
准确定量牙菌斑样本中单个细菌种类的细胞数量对于了解牙周炎的细菌病因学是必要的。实时PCR提供了一种灵敏、高效和可靠的定量方法。使用TaqMan系统，我们能够确定牙菌斑样品中存在的牙龈卟啉单胞菌的量和细菌细胞的总数。使用种特异性引物和荧光探针，从牙龈卟啉单胞菌细胞的系列稀释液中检测DNA在很大的DNA浓度范围内呈线性（相关系数5 0.96）。在单独的牙龈卟啉单胞菌DNA和与从牙菌斑分离的DNA混合的相同DNA之间没有观察到差异，表明牙龈卟啉单胞菌水平可以从临床样品中准确地确定。使用通用引物和荧光探针测定所有细菌物种的细胞总数。使用四种不同的细菌物种的标准曲线得到了类似的结果（相关系数5 - 0.86）。牙龈卟啉单胞菌和总细菌的水平从一系列人牙菌斑样品中确定。在牙周炎受试者的几个样本中观察到高水平的牙龈卟啉单胞菌，而在健康受试者的样本中没有观察到。实时荧光定量PCR检测牙龈卟啉单胞菌是一种灵敏、可靠的方法。此外，它允许测定复杂样品中存在的细菌细胞总数，从而可以测定牙龈卟啉单胞菌细胞的百分比。
Accurate quantitation of the number of cells of individual bacterial species in dental plaque samples is needed for understanding the bacterial etiology of periodontitis. Real-time PCR offers a sensitive, efficient, and reliable approach to quantitation. Using the TaqMan system we were able to determine both the amount of Porphyromonas gingivalis and the total number of bacterial cells present in plaque samples. Using speciesspecific primers and a fluorescent probe, detection of DNA from serial dilutions of P. gingivalis cells was linear over a large range of DNA concentrations (correlation coefficient 5 0.96). No difference was observed between P. gingivalis DNA alone and the same DNA mixed with DNA isolated from dental plaque, indicating that P. gingivalis levels can be determined accurately from clinical samples. The total number of cells of all bacterial species was determined using universal primers and a fluorescent probe. Standard curves using four different bacterial species gave similar results (correlation coefficient 5 0.86). Levels of both P. gingivalis and total bacteria were determined from a series of human plaque samples. High levels of P. gingivalis were observed in several of the samples from subjects with periodontitis and none of those from healthy subjects. Real-time quantitative PCR provided a sensitive and reliable method for quantitating P. gingivalis. In addition, it allowed the determination of the total number of bacterial cells present in a complex sample so that the percentage of P. gingivalis cells could be determined.
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影响因子：
6.7
作者：
Tanner, A;Maiden, MFJ;Kent, RL
通讯作者：
Kent, RL
影响因子：
6.7
作者：
Socransky, SS;Haffajee, AD;Kent, RL
通讯作者：
Kent, RL
患有牙龈炎或牙周炎的老年受试者的临床和微生物学发现。
影响因子：
6.7
作者：
Schlegel-Bregenzer,B;Persson,RE;Lukehart,S;Braham,P;Oswald,T;Persson,GR
通讯作者：
Persson,GR
影响因子：
6.7
作者：
Haffajee, AD;Cugini, MA;Socransky, SS
通讯作者：
Socransky, SS