Lacroix J M, Walker C B
Periodontal Disease Research Center, College of Dentistry, University of Florida, Gainesville, USA.
Oral Microbiol Immunol. 1996 Aug;11(4):282-8. doi: 10.1111/j.1399-302x.1996.tb00182.x.
Subgingival plaque samples were collected from 68 patients with a history of moderate to severe adult periodontitis and enumerated on Trypticase-soy blood agar plates, with and without tetracycline at 4 micrograms/ml. Each different colony morphotype was enumerated, and a representative colony was subcultured for identification and examined for the tetracycline resistance gene tet(Q) by polymerase chain reaction (PCR) amplification and DNA hybridization, using a fragment of tetA(Q)2 from Bacteroides fragilis 1126. PCR primers (5'-GGCTTCTACGACATCTATTA-3' and 5'-CATCAACATTTATCTCTCTG-3') were chosen to amplify a 755 bp region of tet(Q). The subgingival plaque samples were also tested by PCR. Approximately 12% of the total cultivable flora was resistant to tetracycline, and the percentage of the tetracycline-resistant cultivable flora with the tet(Q) gene varied greatly from one patient to another with a range from 0.0 to 67%. Half of the 68 subgingival plaque samples were positive or weakly positive for tet(Q) by PCR. Approximately 15% of the 210 isolates subcultured with resistance to tetracycline, (> or = 4 micrograms/ml) contained tet(Q), and 60% contained tet(M). All of the tet(Q)-resistant isolates were gram-negative anaerobic bacilli and included all of the Prevotella and Bacteroides isolates.
从68例有中度至重度成人牙周炎病史的患者中采集龈下菌斑样本,并在含和不含4微克/毫升四环素的胰蛋白胨大豆血琼脂平板上进行计数。对每种不同的菌落形态型进行计数,并选取一个代表性菌落进行传代培养以进行鉴定,并通过聚合酶链反应(PCR)扩增和DNA杂交,使用脆弱拟杆菌1126的tetA(Q)2片段检测四环素抗性基因tet(Q)。选择PCR引物(5'-GGCTTCTACGACATCTATTA-3'和5'-CATCAACATTTATCTCTCTG-3')扩增tet(Q)的755 bp区域。龈下菌斑样本也通过PCR进行检测。大约12%的可培养菌群对四环素耐药,携带tet(Q)基因的四环素耐药可培养菌群的百分比在不同患者之间差异很大,范围从0.0%到67%。68个龈下菌斑样本中有一半通过PCR检测tet(Q)呈阳性或弱阳性。在对四环素耐药(≥4微克/毫升)的210株传代培养分离株中,约15%含有tet(Q),60%含有tet(M)。所有携带tet(Q)的耐药分离株均为革兰氏阴性厌氧杆菌,包括所有普雷沃菌属和拟杆菌属分离株。