Macheleidt A, Müller H P, Eger T, Putzker M, Zöller L
Central Institute of the German Armed Forces Medical Service, Koblenz, Germany.
J Periodontal Res. 1999 May;34(4):179-87. doi: 10.1111/j.1600-0765.1999.tb02240.x.
Actinobacillus actinomycetemcomitans is a major periodontal pathogen which is associated with both early-onset periodontitis and adult cases refractory to conventional periodontal therapy, although the organism has also been shown to be widely distributed among dentate healthy individuals. The observed disease status may be associated with a variation in virulence of different strains or clones. The aim of the present study was to analyse genotype distribution as assessed by an arbitrarily primed polymerase chain reaction (AP-PCR) among 51 isolates of A. actinomycetemcomitans recovered from more than 200 young adult recruits with no or minor periodontal disease. In addition, isolates from 25 periodontitis patients as well as reference strains were genotyped. Primers amplifying (i) a specific sequence in the ltxA region, (ii) a specific 16S rRNA sequence and (iii) sequences in the leukotoxin promoter region were used to verify species identity of the strains. Three random oligonucleotide primers were employed to analyse genomic polymorphisms of the organism by means of PCR. A total of 19 genotypes could be distinguished, which were grouped by cluster analysis into 5 major clusters based on genetic similarity and a complete linkage sort. Whereas 3 clusters assembled A. actinomycetemcomitans genotypes isolated from both healthy subjects and periodontitis patients, one cluster containing 4 different genotypes exclusively comprised isolates from healthy or gingivitis subjects. Another cluster with 2 genotypes consisted of strains originating from periodontitis patients (p < 0.05). One strain characterized by a specific 530 bp deletion in the promoter region of the leukotoxin region was identified in a Ghanese patient with localized juvenile periodontitis. It was concluded that there is considerable clonal diversity of A. actinomycetemcomitans strains isolated from healthy or periodontally diseased subjects, and that genetically closely related groups might be associated with health or disease.
伴放线放线杆菌是一种主要的牙周病原体，与早发性牙周炎以及对传统牙周治疗无效的成人牙周炎病例均有关联，尽管该微生物也已被证明在有牙健康个体中广泛分布。观察到的疾病状态可能与不同菌株或克隆的毒力差异有关。本研究的目的是通过任意引物聚合酶链反应（AP-PCR）分析从200多名无或仅有轻微牙周疾病的年轻成年新兵中分离出的51株伴放线放线杆菌的基因型分布。此外，对来自25名牙周炎患者的分离株以及参考菌株进行了基因分型。使用扩增（i）ltxA区域特定序列、（ii）特定16S rRNA序列和（iii）白细胞毒素启动子区域序列的引物来验证菌株的种属身份。使用三种随机寡核苷酸引物通过PCR分析该微生物的基因组多态性。总共可区分出19种基因型，通过聚类分析根据遗传相似性和完全连锁分类将其分为5个主要聚类。其中3个聚类汇集了从健康受试者和牙周炎患者中分离出的伴放线放线杆菌基因型，一个包含4种不同基因型的聚类仅由来自健康或牙龈炎受试者的分离株组成。另一个有2种基因型的聚类由源自牙周炎患者的菌株组成（p<0.05）。在一名患有局限性青少年牙周炎的加纳患者中鉴定出一株在白细胞毒素区域启动子区域有特定530 bp缺失的菌株。得出的结论是，从健康或牙周疾病受试者中分离出的伴放线放线杆菌菌株存在相当大的克隆多样性，并且遗传关系密切的组可能与健康或疾病有关。
