Gushchin A E, Taganov K D, Topol' Iu Iu, Govorun V M
Mol Gen Mikrobiol Virusol. 1998(4):37-40.
Single-strand conformation polymorphism (SSCP) method was used for nucleotide sequence variation analysis of the gyrase A subunit quinolone resistance determining region (gyrA QRDR) in a laboratory and clinical strains of M. hominis. The couple of primers selected for this region amplified specific product in clinical material. M. hominis cultures growing in the presence of different concentrations of ciprofloxacin were studied by the SSCP method. Ser(83) to Leu mutation described previously was detected in the presence of quinolone in concentrations of at least 10 mcg/ml. In addition, 11 clinical samples were tested. In all cases the results of SSCP were confirmed by direct sequencing of the region. In 2 cases the sequences of gyrA QRDR in clinical strains were the same as in the laboratory strain. A Ser(83)-Leu mutation was identified in 1 clinical sample, while in others nucleotide substitutes did not lead to changes in amino acid sequences. These data demonstrate high informative value of the SSCP method for evaluating nucleotide variation in gyrA QRDR and quinolone resistance of M. hominis.
采用单链构象多态性(SSCP)方法对人型支原体实验室菌株和临床菌株的gyrase A亚基喹诺酮耐药决定区(gyrA QRDR)进行核苷酸序列变异分析。针对该区域选择的一对引物在临床材料中扩增出特异性产物。通过SSCP方法研究了在不同浓度环丙沙星存在下生长的人型支原体培养物。在喹诺酮浓度至少为10 mcg/ml的情况下,检测到先前描述的Ser(83)至Leu突变。此外,对11份临床样本进行了检测。在所有情况下,SSCP结果均通过该区域的直接测序得到证实。在2例临床菌株中,gyrA QRDR序列与实验室菌株相同。在1份临床样本中鉴定出Ser(83)-Leu突变,而在其他样本中核苷酸替代未导致氨基酸序列改变。这些数据表明SSCP方法在评估人型支原体gyrA QRDR核苷酸变异和喹诺酮耐药性方面具有很高的信息价值。