Canu Annie, Abbas Ahmed, Malbruny Brigitte, Sichel François, Leclercq Roland
UFR des Sciences Pharmaceutiques, Groupe Régional d'Etudes sur le Cancer, Université de Caen/Basse-Normandie, France.
Antimicrob Agents Chemother. 2004 Jan;48(1):297-304. doi: 10.1128/AAC.48.1.297-304.2004.
Mutations in genes coding for L4 (rplD) or L22 (rplV) ribosomal proteins or in 23S rRNA (rrl gene) are reported as a cause of macrolide resistance in streptococci and staphylococci. This study was aimed at evaluating a denaturing high-performance liquid chromatography (DHPLC) technique as a rapid mutation screening method. Portions of these genes were amplified by PCR from total DNA of 48 strains of Streptococcus pneumoniae (n = 22), Staphylococcus aureus (n = 16), Streptococcus pyogenes (n = 6), Streptococcus oralis (n = 2), and group G streptococcus (n = 2). Thirty-seven of these strains were resistant to macrolides and harbored one or several mutations in one or two of the target genes, and 11 were susceptible. PCR products were analyzed by DHPLC. All mutations were detected, except a point mutation in a pneumococcal rplD gene. The method detected one mutated rrl copy out of six in S. aureus. This automated method is promising for screening of mutations involved in macrolide resistance in gram-positive cocci.
据报道，编码L4（rplD）或L22（rplV）核糖体蛋白的基因或23S rRNA（rrl基因）中的突变是链球菌和葡萄球菌对大环内酯类耐药的原因。本研究旨在评估变性高效液相色谱（DHPLC）技术作为一种快速突变筛选方法。通过PCR从48株肺炎链球菌（n = 22）、金黄色葡萄球菌（n = 16）、化脓性链球菌（n = 6）、口腔链球菌（n = 2）和G组链球菌（n = 2）的总DNA中扩增这些基因的部分片段。其中37株对大环内酯类耐药，在一个或两个靶基因中存在一个或几个突变，11株敏感。通过DHPLC分析PCR产物。除了肺炎链球菌rplD基因中的一个点突变外，所有突变均被检测到。该方法在金黄色葡萄球菌中检测到六个rrl拷贝中有一个发生突变。这种自动化方法有望用于筛选革兰氏阳性球菌中与大环内酯类耐药相关的突变。
