Siqueira José F, Rôças Isabela N
Department of Endodontics, Faculty of Dentistry, Estácio de Sá University, R. Herotides de Oliveira 61/601, Icarai, Niterói, RJ 24230-230, Rio de Janeiro, Brazil.
J Dent. 2003 Jul;31(5):333-9. doi: 10.1016/s0300-5712(03)00051-4.
This paper reviews the principles of polymerase chain reaction (PCR) methodology, its application in identification of endodontic pathogens and the perspectives regarding the knowledge to be reached with the use of this highly sensitive, specific and accurate methodology as a microbial identification test.
Studies published in the medical, dental and biological literature.
Evaluation of published epidemiological studies examining the endodontic microbiota through PCR methodology.
PCR technology has enabled the detection of bacterial species that are difficult or even impossible to culture as well as cultivable bacterial strains showing a phenotypically divergent or convergent behaviour. Moreover, PCR is more rapid, much more sensitive, and more accurate when compared with culture. Its use in endodontics to investigate the microbiota associated with infected root canals has expanded the knowledge on the bacteria involved in the pathogenesis of periradicular diseases. For instance, Tannerella forsythensis (formerly Bacteroides forsythus), Treponema denticola, other Treponema species, Dialister pneumosintes, and Prevotella tannerae were detected in infected root canals for the first time and in high prevalence when using PCR analysis. The diversity of endodontic microbiota has been demonstrated by studies using PCR amplification, cloning and sequencing of the PCR products. Moreover, other fastidious bacterial species, such as Porphyromonas endodontalis, Porphyromonas gingivalis and some Eubacterium spp., have been reported in endodontic infections at a higher prevalence than those reported by culture procedures.
本文综述聚合酶链反应(PCR)方法的原理、其在牙髓病病原体鉴定中的应用,以及关于将这种高度灵敏、特异且准确的方法用作微生物鉴定测试所获得知识的展望。
医学、牙科和生物学文献中发表的研究。
对通过PCR方法检测牙髓微生物群的已发表流行病学研究进行评估。
PCR技术能够检测出难以培养甚至无法培养的细菌种类,以及表现出表型趋异或趋同行为的可培养细菌菌株。此外,与培养法相比,PCR更为快速、灵敏且准确。其在牙髓病学中用于研究与感染根管相关的微生物群,扩展了对根尖周病发病机制中涉及细菌的认识。例如,使用PCR分析首次在感染根管中高频率检测到福赛坦纳菌(以前称为福赛拟杆菌)、具核梭杆菌、其他梭杆菌属物种、嗜沫栖牙菌和坦纳普雷沃菌。通过对PCR产物进行扩增、克隆和测序的研究已证明牙髓微生物群的多样性。此外,其他苛养菌,如牙髓卟啉单胞菌、牙龈卟啉单胞菌和一些真杆菌属物种,在牙髓感染中的检出率高于培养法报告的结果。