Department of Pathology and Propedeutics, Araçatuba Dental School, São Paulo State University, Araçatuba, SP, Brazil.
J Appl Oral Sci. 2009 Sep-Oct;17(5):440-5. doi: 10.1590/s1678-77572009000500017.
Aggregatibacter actinomycetemcomitans is associated with periodontal disease, especially localized aggressive periodontitis, produces a potent leukotoxin and its distribution is influenced by ethnic characteristics of the population.
Using culture and polymerase chain reaction (PCR) techniques, this study evaluated the occurrence of this microorganism and the distribution of leukotoxic strains isolated from Indians belonging to the Umutima Reservation, Mato Grosso, Brazil.
Forty-eight native Brazilians with gingivitis and 38 with chronic periodontitis, belonging to Umutina, Paresi, Bororo, Bakairi, Kayabi, Irantxe, Nambikwara and Terena ethnicities, were studied. Subgingival, supragingival and saliva samples of each patient were collected and transferred to VMGA III medium and to ultra pure Milli Q water. Bacteria were grown on TSBV agar and incubated in anaerobiosis (90% N2 + 10% CO2) at 37 degrees C for 72 h. The presence of the ltx promoter was determined by PCR, and a 530 bp deletion in the promoter was evaluated by using specific primers.
A. actinomycetemcomitans was isolated from 8.33% of saliva, supragingival and subgingival samples from patients with gingivitis and from 18.42% of saliva and supragingival biofilm, and 26.32% subgingival biofilm from patients with chronic periodontitis. By PCR, the bacterial DNA was detected in 8.33% of saliva, supragingival and subgingival biofilms from patients with gingivitis and from 23.68% of saliva, 28.95% supragingival biofilm and 34.21% subgingival biofilm from patients with periodontitis. All strains were grouped as non-JP2 clones based on the absence of deletion in the leukotoxin promoter. Differences among the microbial and clinical parameters in patients were analyzed by using the Mann-Whitney, Chi-square or Fisher's exact tests.
The present results suggest that A. actinomycetemcomitans can be related to the attachment loss in this population, but the presence of minimally leukotoxic strains, as well as its role in the pathogenesis of the periodontitis in these native Brazilians need to be further investigated.
本研究采用培养和聚合酶链反应(PCR)技术,评估巴西乌穆蒂马保留地印第安人群中,牙周病特别是局限性侵袭性牙周炎患者携带和分离出的具白细胞毒素活性的聚集放线杆菌(Aggregatibacter actinomycetemcomitans)的分布情况。
共检测 48 例患有牙龈炎和 38 例慢性牙周炎的巴西印第安人,他们分别属于乌穆蒂纳(Umutina)、帕雷西(Paresi)、博洛罗(Bororo)、巴凯瑞(Bakairi)、卡亚比(Kayabi)、伊兰泰克(Irantxe)、南比克瓦拉(Nambikwara)和特雷纳(Terena)等族群。每位患者的龈下、龈上和唾液样本都采集并分别转移到 VMGA III 培养基和超纯 Milli Q 水中。将细菌接种于 TSBV 琼脂平板并在 37℃下于厌氧(90% N2 + 10% CO2)环境中孵育 72 小时。采用 PCR 检测白细胞毒素启动子的存在,并用特定引物评估启动子中 530bp 缺失情况。
从牙龈炎患者的龈上和龈下样本、唾液样本中分别分离出 8.33%、8.33%和 18.42%的 A. actinomycetemcomitans,从慢性牙周炎患者的唾液和龈上生物膜样本中分别分离出 26.32%和 18.42%的 A. actinomycetemcomitans,从慢性牙周炎患者的龈下生物膜样本中分离出 23.68%的 A. actinomycetemcomitans。PCR 检测到牙龈炎患者的龈上和龈下生物膜样本、唾液样本中分别有 8.33%、8.33%和 23.68%的细菌 DNA,慢性牙周炎患者的唾液、龈上生物膜和龈下生物膜样本中分别有 28.95%、28.95%和 34.21%的细菌 DNA。根据白细胞毒素启动子缺失情况,所有菌株均被归类为非 JP2 克隆。采用曼-惠特尼(Mann-Whitney)、卡方或 Fisher 精确检验对患者微生物和临床参数进行分析。
本研究结果提示,A. actinomycetemcomitans 可能与该人群的附着丧失有关,但需要进一步研究这些巴西印第安人群中低白细胞毒素活性菌株的存在及其在牙周炎发病机制中的作用。