Amano A, Nakagawa I, Kataoka K, Morisaki I, Hamada S
Division of Special Care Dentistry, Osaka University Faculty of Dentistry, Suita-Osaka 565-0871, Japan.
J Clin Microbiol. 1999 May;37(5):1426-30. doi: 10.1128/JCM.37.5.1426-1430.1999.
Fimbriae (FimA) of Porphyromonas gingivalis are filamentous components on the cell surface and are thought to play an important role in the colonization and invasion of periodontal tissues. We previously demonstrated that fimA can be classified into four variants (types I to IV) on the basis of the nucleotide sequences of the fimA gene. In the present study, we attempted to detect the four different fimA genes in saliva and plaque samples isolated from patients with periodontitis using the PCR method. Four sets of fimA type-specific primers were designed for the PCR assay. These primers selectively amplified 392-bp (type I), 257-bp (type II), 247-bp (type III), and 251-bp (type IV) DNA fragments of the fimA gene. Positive PCR results were observed with reference strains of P. gingivalis in a type-specific manner. All other laboratory strains of oral and nonoral bacteria gave negative results. The sensitivity of the PCR assay for fimA type-specific detection was between 5 and 50 cells of P. gingivalis. Clinical samples were obtained from saliva and subgingival plaque from deep pockets (>/=4 mm) of 93 patients with periodontitis. Bacterial genomic DNA was isolated from the samples, and the targeted fragments were amplified by PCR. The presence of P. gingivalis was demonstrated in 73 patients (78.5%), and a single fimA gene was detected in most patients. The distribution of the four fimA types among the P. gingivalis-positive patients was as follows: type I, 5.4%; type II, 58.9%; type III, 6. 8%; type IV, 12.3%; types I and II, 6.8%; types II and IV, 2.7%; and untypeable, 6.8%. P. gingivalis with type II fimA was detected more frequently in the deeper pockets, and a significant difference of the occurrence was observed between shallow (4 mm) and deep (>/=8 mm) pockets. These results suggest that P. gingivalis strains that possess type II fimA are significantly more predominant in periodontitis patients, and we speculate that these organisms are involved in the destructive progression of periodontal diseases.
牙龈卟啉单胞菌的菌毛(FimA)是细胞表面的丝状成分,被认为在牙周组织的定植和侵袭中起重要作用。我们之前证明,基于fimA基因的核苷酸序列,fimA可分为四种变体(I型至IV型)。在本研究中,我们试图使用PCR方法检测从牙周炎患者分离的唾液和菌斑样本中的四种不同fimA基因。为PCR检测设计了四组fimA型特异性引物。这些引物选择性地扩增了fimA基因的392 bp(I型)、257 bp(II型)、247 bp(III型)和251 bp(IV型)DNA片段。牙龈卟啉单胞菌参考菌株以型特异性方式观察到PCR阳性结果。所有其他口腔和非口腔细菌的实验室菌株均给出阴性结果。fimA型特异性检测的PCR检测灵敏度为5至50个牙龈卟啉单胞菌细胞。从93例牙周炎患者的唾液和深牙周袋(≥4 mm)的龈下菌斑中获取临床样本。从样本中分离细菌基因组DNA,并通过PCR扩增目标片段。73例患者(78.5%)检测到牙龈卟啉单胞菌,大多数患者检测到单个fimA基因。牙龈卟啉单胞菌阳性患者中四种fimA型的分布如下:I型,5.4%;II型,58.9%;III型,6.8%;IV型,12.3%;I型和II型,6.8%;II型和IV型,2.7%;无法分型,6.8%。II型fimA的牙龈卟啉单胞菌在较深牙周袋中检测更为频繁,浅袋(4 mm)和深袋(≥8 mm)之间的发生率存在显著差异。这些结果表明,具有II型fimA的牙龈卟啉单胞菌菌株在牙周炎患者中明显更为占优势,我们推测这些微生物参与了牙周疾病的破坏性进展。