Lotufo R F, Flynn J, Chen C, Slots J
Disciplina de Periodontia, Faculdade de Odontologia, Universidade de São Paulo, Brazil.
Oral Microbiol Immunol. 1994 Jun;9(3):154-60. doi: 10.1111/j.1399-302x.1994.tb00052.x.
The usefulness of a digoxigenin-labeled genomic DNA probe for the detection of subgingival Bacteroides forsythus was examined. In addition, the arbitrarily primed polymerase chain reaction (AP-PCR) was used to delineate the genetic diversity of B. forsythus periodontal isolates. The DNA probe detected 10(3) B. forsythus cells and yielded a strong signal at 10(4) cells. It reacted with B. forsythus ATCC 43037T and 44 clinical isolates and showed no detectable reactivity with 75 strains of 24 other oral microbial species. In comparison to culture, the DNA probe in a dot-blot method demonstrated a sensitivity of 88.8% and a specificity of 38.4% (accuracy, 72.5%). By colony-blotting on primary plates, a sensitivity of 98.1% and a specificity of 53.8% (accuracy, 82.5%) were obtained. B. forsythus was detected in 449 (73.1%) of 614 periodontitis patients. The occurrence of the organism was closely associated with Porphyromonas gingivalis, both species being present in 54.8% and absent in 22.2% of 270 study samples. AP-PCR identified 24 B. forsythus genotypes among 27 test strains. This study demonstrated the utility of a non-radioactive genomic probe for direct detection of B. forsythus in subgingival specimens. The species showed a considerable degree of genetic diversity. DNA analysis may help to determine the role of B. forsythus in periodontal disease and its mode of transmission among exposed individuals.
研究了地高辛配体标记的基因组DNA探针用于检测龈下福赛坦氏菌的效用。此外,还采用了随机引物聚合酶链反应(AP-PCR)来描绘牙周分离出的福赛坦氏菌的遗传多样性。该DNA探针能检测到10³个福赛坦氏菌细胞,并在10⁴个细胞时产生强信号。它与福赛坦氏菌ATCC 43037T及44株临床分离株发生反应,而与其他24种口腔微生物的75株菌株未显示出可检测到的反应性。与培养法相比,斑点印迹法中的DNA探针灵敏度为88.8%,特异性为38.4%(准确率为72.5%)。通过在原代平板上进行菌落印迹,灵敏度为98.1%,特异性为53.8%(准确率为82.5%)。在614例牙周炎患者中,有449例(73.1%)检测到福赛坦氏菌。该菌的出现与牙龈卟啉单胞菌密切相关,在270份研究样本中,两种菌同时存在的占54.8%,均不存在的占22.2%。AP-PCR在27株测试菌株中鉴定出24种福赛坦氏菌基因型。本研究证明了非放射性基因组探针在直接检测龈下标本中福赛坦氏菌方面的效用。该菌种显示出相当程度的遗传多样性。DNA分析可能有助于确定福赛坦氏菌在牙周病中的作用及其在暴露个体间的传播方式。