Slots J, Ashimoto A, Flynn M J, Li G, Chen C
University of Southern California School of Dentistry, Los Angeles 90089-0641, USA.
Clin Infect Dis. 1995 Jun;20 Suppl 2:S304-7. doi: 10.1093/clinids/20.supplement_2.s304.
The utility of the 16S ribosomal RNA-based polymerase chain reaction (PCR) for detection of Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Campylobacter rectus, Eikenella corrodens, Porphyromonas gingivalis, Prevotella intermedia, and Treponema denticola was examined and compared with that of anaerobic culture. Primer pairs consisting of 20-27 nucleotides amplified 404- to 688-bp regions of 16S ribosomal RNA genes of these organisms. This method had a lower detection limit of 50 target cells in a background of 10(7) cells. Its specificity for B. forsythus, P. gingivalis, and T. denticola seemed high. The primers for A. actinomycetemcomitans, C. rectus, and P. intermedia cross-reacted with some closely related species but did not reveal amplification products in tests with more distantly related organisms. The primers for E. corrodens did not seem to cross-react with oral organisms. This PCR technique was sensitive, reproducible, and easy to perform. PCR-based amplification may prove valuable for the detection of some periodontal pathogens in crude subgingival specimens.
研究了基于16S核糖体RNA的聚合酶链反应（PCR）用于检测伴放线放线杆菌、福赛斯坦纳菌、直肠弯曲菌、腐蚀艾肯菌、牙龈卟啉单胞菌、中间普氏菌和具核梭杆菌的效用，并与厌氧培养法进行了比较。由20 - 27个核苷酸组成的引物对扩增了这些微生物16S核糖体RNA基因的404至688碱基对区域。该方法在10⁷个细胞的背景下检测下限为50个靶细胞。其对福赛斯坦纳菌、牙龈卟啉单胞菌和具核梭杆菌的特异性似乎较高。伴放线放线杆菌、直肠弯曲菌和中间普氏菌的引物与一些密切相关的菌种发生交叉反应，但在与亲缘关系较远的微生物的检测中未显示扩增产物。腐蚀艾肯菌的引物似乎未与口腔微生物发生交叉反应。这种PCR技术灵敏、可重复且易于操作。基于PCR的扩增对于在龈下粗标本中检测某些牙周病原体可能具有重要价值。
