王嘉尔击剑朋友:哪位高手知道Box－PCR的信息？谢谢了！！！

肺炎链球菌临床菌株Tn1545和Tn917介导的大环内酯类耐药性研究

吴文娟 洪秀华 张泓 蒋燕群 陆权 倪语星

摘 要：目的探讨上海地区大环内酯类耐药肺炎链球菌ermB基因表达及传播特性.方法收集上海地区对红霉素耐药的肺炎链球菌共86株,用E试验和K-B纸片扩散法检测其对12种抗生素的敏感性;用双纸片法(D试验)确定大环内酯类耐药表型;用PCR 扩增检测耐药基因ermB、mefE和Tn1545、Tn917中ermB基因调控序列,经测序、BLAST比对分析调控序列多态性并按菌株携带Tn1545和Tn917的不同分为Tn1545组和Tn917组,分析二者介导的耐药表型的不同;用BOX-PCR分析不同菌株遗传背景.结果 (1)86株红霉素耐药肺炎链球菌中ermB、mefE、Tn1545、Tn917检出率分别为94%、46%、87%、7%,基因型为ermB-mefE+的5个菌株未检出Tn1545和Tn917.(2)Tn1545组和Tn917组Sp大多对红霉素高度耐药(最低抑菌浓度MIC50 256 μg/ml),Tn917组对β内酰胺类抗生素的MIC低于Tn1545组,对四环素、复方磺胺及左氧氟沙星耐药率前者亦低于后者.(3)Tn917组中三株菌发生ermB基因启动子区194 bp片段的删除和前导肽N末端TAAA插入,可能导致ermB基因由诱导型转变成组成型表达;Tn1545组菌株主要表现为cMLSB表型.(4)BOX-PCR图谱呈散发性,未发现单一的耐药克隆株流行. 结论上海地区大环内酯类耐药肺炎链球菌主要是由ermB基因介导的高水平、组成型耐药,ermB基因大多由Tn1545携带并水平转移传播.
关键词：链球菌,肺炎;抗生素,大环内酯;DNA可移植因子;耐药性,多药

Characterization of macrolide-resistance mediated by transposable elements Tn1545 and Tn917 in clinical Isolates of Streptococcus pneumoniae

WU Wen-juan HONG Xiu-hua ZHANG Hong JIANG Yan-qun LU Quan NI Yu-xing

作者简介：倪语星,Email:yuxing_ni@yahoo.com.cn,通讯作者
作者单位：吴文娟（200025,上海交通大学医学院附属瑞金医院临床微生物科）
洪秀华（上海交通大学医学院检验医学系）
张泓（上海交通大学附属儿童医院检验科）
蒋燕群（上海交通大学附属第六人民医院检验科）
陆权（上海交通大学附属儿童医院呼吸科）
倪语星（200025,上海交通大学医学院附属瑞金医院临床微生物科）

参考文献：

[1]王辉,俞云松,刘勇.2002-2003年中国社区呼吸道感染常见病原菌的耐药性监测.中华结核和呼吸杂志,2004,27:155-160.
[2]姚开虎,陆权,邓力,等.2000-2002年北京、上海和广州儿童肺炎链球菌携带及抗生素耐药性监测.中华医学杂志,2005,85:1957-1961.
[3]Shaw JH,Clewell DB.Complete nucleotide sequence of macrolide-lincosamide-streptogramin B resistance transposon Tn917 in Streptococcus faecalis.J Bacteriol,1985,164:782-796.
[4]Trieu-Cuot P,Poyart-Salmeron C,Carlier C,et al.Nucleotide sequence of the erythromycin resistance gene of the conjugative transposon Tn1545.Nucleic Acids Res,1990,18:3660.
[5]Marchese A,Tonoli E,Debbia EA,et a1.Macrolide resistance mechanisms and expression of phenotypes among Streptococcus pneumoniae circulating in Italy.J Antimicrob Chemother,1999,44:461-464.
[6]Sutcliffe J,Grebe T,Tait-Kamradt A.Detection of erythromycin-resistant determinants by PCR.Antimicrob Agents Chemother,1996,40:2562-2566.
[7]Daly MM,Doktor S,Flamm R,et a1.Characterization and prevalence of MefA,MefE,and the associated msr(D) gene in Streptococcus pneumoniae clinical isolates.J Clin Microbiol,2004,42:3570-3574.
[8]van Belkum A,Sluijuter M,de Groot R,et al.Novel BOX repeat PCR assay for high-resolution typing of Streptococcus pneumoniae strains.J Clin Microbiol,1996,34:1176-1179.
[9]Okitsu N,Kaieda S,Yano H,et al.Characterization of ermB gene transposition by Tn1545 and Tn917 in macrolide-resistant Streptococcus pneumoniae isolates.J Clin Microbiol,2005,43:168-173.
[10]周宏,邓力,叶启慈.广州地区ermB和mef E基因介导对红霉素耐药的肺炎链球菌及其耐药性分析.中华检验医学杂志,2004,27:111-113.
[11]McDougal LK,Tenover FC,Lee LN,et al.Detection of Tn917-like sequences within a Tn916-like conjugative transposon (Tn3872) in erythromycin-resistant isolates of Streptococcus pneumoniae.Antimicrob Agents Chemother,1998,42:2312-2318.
[12]Leclercq R,Courvalin P.Resistance to macrolides and related antibiotics in Streptococcus pneumoniae.Antimicrob Agents Chemother,2002,46:2727-2734.
[13]Oh TG,Kwon AR,Choi EC.Induction of ermAMR from a clinical strain of Enterococcus faecalis by 16-membered-ring macrolide antibiotics.J Bacteriol,1998,180:5788-5791.
[14]Rosato A,Vicarini H,Leclercq R.Inducible or constitutive expression of resistance in clinical isolates of streptococci and enterococci cross-resistant to erythromycin and lincomycin.J Antimicrob Chemother,1999,43:559-562.
[15]Zhong P,Cao Z,Hammond R,et al.Induction of ribosome methylation in MLS-resistant streptococcus pneumoniae by macrolides and ketolides.Microb Drug Resist,1999,5:183-188.

我电脑上不了外网，但在google上找到了英文解释。你能上外网就自己看看吧。学生物的，英文应该没问题。呵呵
http://www.changbioscience.com/primo/pcr/eBOXsPCR.htm