Malheiros Veruska de João, Avila-Campos Mario Julio
Laboratório de Anaeróbios, Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, SP, Brazil.
Rev Saude Publica. 2004 Oct;38(5):723-8. doi: 10.1590/s0034-89102004000500016. Epub 2004 Oct 18.
To comparatively detect A. actinomycetemcomitans and F. nucleatum from periodontal and healthy sites.
Subgingival clinical samples from 50 periodontitis adult patients and 50 healthy subjects were analyzed. Both organisms were isolated using a trypticase soy agar-bacitracin-vancomycin (TSBV) medium and detected by PCR. Conventional biochemical tests were used for bacteria identification.
A. actinomycetemcomitans and F. nucleatum were isolated in 18% and 20% of the patients, respectively, and in 2% and 24% of healthy subjects. Among A. actinomycetemcomitans isolates, biotype II was the most prevalent. Primer pair AA was 100% sensitive in the detection of A. actinomycetemcomitans from both subject groups. Primers ASH and FU were also 100% sensitive to detect this organism in healthy subject samples. Primer pair FN5047 was more sensitive to detect F. nucleatum in patients or in healthy samples than primer 5059S. Primers ASH and 5059S were more specific in the detection of A. actinomycetemcomitans and F. nucleatum, respectively, in patients and in healthy subject samples.
PCR is an effective tool for detecting periodontal pathogens in subgingival samples, providing a faster and safer diagnostic tool of periodontal diseases. The method's sensitivity and specificity is conditioned by the choice of the set of primers used.
比较检测牙周部位和健康部位的伴放线放线杆菌和具核梭杆菌。
对50例成年牙周炎患者和50例健康受试者的龈下临床样本进行分析。使用胰蛋白酶大豆琼脂-杆菌肽-万古霉素(TSBV)培养基分离这两种微生物,并通过聚合酶链反应(PCR)进行检测。采用常规生化试验进行细菌鉴定。
伴放线放线杆菌和具核梭杆菌在患者中的分离率分别为18%和20%,在健康受试者中的分离率分别为2%和24%。在分离出的伴放线放线杆菌中,生物型II最为常见。引物对AA在检测两个受试组中的伴放线放线杆菌时敏感性为100%。引物ASH和FU在检测健康受试者样本中的该微生物时敏感性也为100%。引物对FN5047在检测患者或健康样本中的具核梭杆菌时比引物5059S更敏感。引物ASH和5059S分别在检测患者和健康受试者样本中的伴放线放线杆菌和具核梭杆菌时更具特异性。
PCR是检测龈下样本中牙周病原体的有效工具,为牙周疾病提供了一种更快、更安全的诊断工具。该方法的敏感性和特异性取决于所用引物组的选择。