Institute of Clinical Microbiology, Faculty of Medicine, University of Szeged, H-6725 Szeged, Semmelweis u 6., Hungary.
Anaerobe. 2010 Jun;16(3):283-8. doi: 10.1016/j.anaerobe.2010.02.005. Epub 2010 Feb 26.
To culture facultative and strict anaerobic bacteria is a well-established method for analyzing subgingival plaque samples. Micro-IDent and micro-IDent Plus (HAIN Lifescience GmbH, Nehren, Germany) tests are two commercially available rapid PCR-based methods for the identification and quantification of putative periodontopathogen bacteria. In this study, we compared these commercial PCR-based hybridization methods with conventional anaerobic culture technique. A total of 36 subgingival plaque samples were collected from periodontal pockets of pregnant women with chronic localized periodontitis. Aliquots of these samples were evaluated with species-specific probes provided by micro-IDent and micro-IDent Plus tests simultaneously, and from the same samples anaerobic and capnophylic bacteria were cultured on selective media. The overall agreement between both methods was excellent for Eubacterium nodatum, Tannerella forsythia and Porphyromonas gingivalis (97-92%), fair for Capnocytophaga sp, Eikenella corrodens, Actinobacillus actinomycetemcomitans, and Prevotella intermedia (91-89%) and poor for Fusobacterium nucleatum, Parvimonas micra (Micromonas micros), and Campylobacter rectus (86-78%). Discrepancies in the results may be explained by inability of culture method to distinguish between closely related taxa (e.i P. intermedia/Prevotella. nigrescens), and problems of keeping periodontopathogen bacteria viable, which is required for successful detection by standard culture method. Nucleic acid-based methods may replace cultivation method as frequently used methods in microbiological diagnosis of progressive periodontitis, thus micro-IDent and micro-IDent Plus tests can be recommended where culture of periodontopathogenic bacteria is not performed in routine microbiology laboratories to analyze subgingival plaque samples.
培养兼性和严格厌氧菌是分析龈下菌斑样本的一种成熟方法。Micro-IDent 和 micro-IDent Plus(德国 HAIN Lifescience GmbH,Nehren)测试是两种市售的基于 PCR 的快速检测方法,可用于鉴定和定量可疑牙周病原菌。在本研究中,我们将这两种商业性基于 PCR 的杂交方法与传统的厌氧培养技术进行了比较。共收集了 36 份来自患有慢性局限性牙周炎的孕妇龈下牙周袋的菌斑样本。这些样本的等分试样同时用 micro-IDent 和 micro-IDent Plus 测试中的种特异性探针进行评估,并且从相同的样本中,在选择性培养基上培养了厌氧和需二氧化碳菌。两种方法之间的总体一致性对于真杆菌属(Eubacterium nodatum)、福赛斯坦纳菌(Tannerella forsythia)和牙龈卟啉单胞菌(Porphyromonas gingivalis)非常好(97-92%),对于噬二氧化碳噬纤维菌(Capnocytophaga sp)、具核梭杆菌(Eikenella corrodens)、伴放线放线杆菌(Actinobacillus actinomycetemcomitans)和中间普氏菌(Prevotella intermedia)为良好(91-89%),而对于核梭杆菌(Fusobacterium nucleatum)、微小微单胞菌(Parvimonas micra)和直肠弯曲菌(Campylobacter rectus)则为较差(86-78%)。结果的差异可能是由于培养方法无法区分密切相关的分类群(例如 P. intermedia/Prevotella. nigrescens),以及保持牙周病原菌存活的问题,这对于标准培养方法的成功检测是必需的。基于核酸的方法可能会取代培养方法,成为进行牙周炎进展性微生物学诊断的常用方法,因此,如果在常规微生物学实验室中未进行牙周病原菌的培养,则可以推荐使用 micro-IDent 和 micro-IDent Plus 测试来分析龈下菌斑样本。