van Steenbergen T J, Timmerman M F, Mikx F H, de Quincey G, van der Weijden G A, van der Velden U, de Graaff J
Department of Oral Microbiology, Academic Centre for Dentistry, Amsterdam, The Netherlands.
J Clin Periodontol. 1996 Oct;23(10):955-9. doi: 10.1111/j.1600-051x.1996.tb00518.x.
The purpose of this study was to compare a commercially available DNA probe technique with conventional cultural techniques for the detection of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Prevotella intermedia in subgingival plaque samples. Samples from 20 patients with moderate to severe periodontitis were evaluated at baseline and during a 15 months period of periodontal treatment. Paperpoints from 4 periodontal pockets per patient were forwarded to Omnigene for DNA probe analysis, and simultaneously inserted paperpoints from the same pockets were analyzed by standard culture techniques. In addition, mixed bacterial samples were constructed harbouring known proportions of 25 strains of A. actinomycetemcomitans, P. gingivalis and P. intermedia each. A relatively low concordance was found between both methods. At baseline a higher detection frequency was found for A. actinomycetemcomitans and P. gingivalis for the DNA probe technique; for P. intermedia the detection frequency by culture was higher. For A. actinomycetemcomitans, 21% of the culture positive samples was positive with the DNA probe. Testing the constructed bacterial samples with the DNA probe method resulted in about 16% false positive results for the 3 species tested. Furthermore, 40% of P. gingivalis strains were not detected by the DNA probe. The present data suggest that at least part of the discrepancies found between the DNA probe technique used and cultural methods are caused by false positive and false negative DNA probe results. Therefore, the value of this DNA probe method for the detection of periodontal pathogens is questionable.
本研究的目的是比较一种市售的DNA探针技术与传统培养技术，用于检测龈下菌斑样本中的伴放线放线杆菌、牙龈卟啉单胞菌和中间普氏菌。对20例中重度牙周炎患者的样本在基线期和为期15个月的牙周治疗期间进行评估。将每位患者4个牙周袋中的纸尖送去进行Omnigene DNA探针分析，同时将来自相同牙周袋的插入纸尖用标准培养技术进行分析。此外，构建了含有已知比例的25株伴放线放线杆菌、牙龈卟啉单胞菌和中间普氏菌的混合细菌样本。两种方法之间的一致性相对较低。在基线期，DNA探针技术检测伴放线放线杆菌和牙龈卟啉单胞菌的频率较高；中间普氏菌的培养检测频率较高。对于伴放线放线杆菌，21%的培养阳性样本用DNA探针检测为阳性。用DNA探针方法检测构建的细菌样本，在所检测的3种细菌中产生了约16%的假阳性结果。此外，40%的牙龈卟啉单胞菌菌株未被DNA探针检测到。目前的数据表明，所使用的DNA探针技术与培养方法之间发现的差异至少部分是由DNA探针的假阳性和假阴性结果引起的。因此，这种DNA探针方法用于检测牙周病原体的价值值得怀疑。
