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无龋和龋活跃个体中变形链球菌的基因型多样性和毒力特征。

Genotypic diversity and virulence traits of Streptococcus mutans in caries-free and caries-active individuals.

作者信息

Napimoga Marcelo Henrique, Kamiya Regianne Umeko, Rosa Rosimeire Takaki, Rosa Edvaldo AntonioR, Höfling José Francisco, de Oliveira Mattos-Graner Renata, Gonçalves Reginaldo Bruno

机构信息

Department of Oral Diagnostics, Faculty of Dentistry of Piracicaba, University of Campinas, Piracicaba, São Paulo, Brazil.

出版信息

J Med Microbiol. 2004 Jul;53(Pt 7):697-703. doi: 10.1099/jmm.0.05512-0.

Abstract

The present study evaluated the relationship between clonal diversity and some virulence traits of Streptococcus mutans isolated from eight caries-free and eight caries-active subjects. A total of 155 S. mutans isolates from caries-free subjects and 144 isolates from caries-active subjects were obtained from samples of saliva, dental plaque and tongue surface and identified by PCR. The isolates were submitted to arbitrarily primed (AP)-PCR (OPA-2 and OPA-13) and multilocus enzyme electrophoresis (MLEE) to establish the genotypic diversity. Production of water-insoluble glucan (WIG) (monitored by SDS-PAGE), final pH of cultures and the ability of bacterial cells to adhere to smooth glass in the presence of sucrose were measured. High and comparable abilities of MLEE and AP-PCR were found to distinguish S. mutans genotypes, using Simpson's index of discrimination (0.971 and 0.968, respectively). The results showed a significant difference (P < 0.01) in the number of genotypes when caries-free and caries-active groups were compared by both fingerprinting methods used. Final pH (P = 0.32) and the percentage of adherence to a glass surface (P = 0.62) did not show differences between the two groups; however, the intensities of WIG bands from the caries-active group were greater than those from the caries-free group (P < 0.01). In addition, WIG was positively correlated with the ability of S. mutans to adhere to a glass surface (r = 0.34, P = 0.02) from caries-active subjects. These data showed that AP-PCR analysis and MLEE are both effective methods for assessing the genetic relatedness of S. mutans. Using these techniques, it was found that there is a larger number of genotypes of S. mutans with increased ability to synthesize WIG in caries-active individuals.

摘要

本研究评估了从8名无龋和8名患龋受试者中分离出的变形链球菌的克隆多样性与某些毒力特性之间的关系。从唾液、牙菌斑和舌面样本中获得了总共155株来自无龋受试者的变形链球菌分离株和144株来自患龋受试者的分离株,并通过聚合酶链反应(PCR)进行鉴定。将这些分离株进行任意引物聚合酶链反应(AP-PCR)(OPA-2和OPA-13)和多位点酶电泳(MLEE)以确定基因型多样性。测量了水不溶性葡聚糖(WIG)的产生(通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)监测)、培养物的最终pH值以及细菌细胞在蔗糖存在下粘附于光滑玻璃的能力。使用辛普森鉴别指数(分别为0.971和0.968)发现,MLEE和AP-PCR区分变形链球菌基因型的能力较高且相当。结果表明,当使用两种指纹图谱方法比较无龋组和患龋组时,基因型数量存在显著差异(P < 0.01)。两组之间的最终pH值(P = 0.32)和玻璃表面粘附百分比(P = 0.62)没有差异;然而,患龋组WIG条带的强度大于无龋组(P < 0.01)。此外,来自患龋受试者的WIG与变形链球菌粘附于玻璃表面的能力呈正相关(r = 0.34,P = 0.02)。这些数据表明,AP-PCR分析和MLEE都是评估变形链球菌遗传相关性的有效方法。使用这些技术发现,在患龋个体中,合成WIG能力增强的变形链球菌基因型数量更多。

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