摘要目的 探讨泌尿生殖道感染的不育症患者精液中分离出的金黄色葡萄球菌对精子活力的影响及其携带的毒力基因谱特征. 方法 收集2009年12月至2011年1月收治的不育症患者精液中分离出的金黄色葡萄球菌89株,以金黄色葡萄球菌ATCC29213为阴性对照菌,采用计算机辅助生殖分析系统Sperm Class Analyzer V.3.2.0,按照WHO(2010版)推荐的精液分析方法,观察89株金黄色葡萄球菌对体检合格的育龄男子正常精子活力的影响,进行精子活动力评级;琼脂稀释法检测喹诺酮类、磺胺类等抗菌药物对实验菌株的最低抑菌浓度(minimum inhibitory concentration,MIC);采用头孢西丁纸片扩散法和mecA基因扩增法确定其中的耐甲氧西林金黄色葡萄球菌(methicillin-resistant Staphylococcus aureus,MRSA);采用多重PCR法对MRSA菌株进行mec耐药基因盒(staphylococcal cassette chromosome mec,SCCmec)分型;采用普通PCR法检测金黄色葡萄球菌的毒力基因. 结果 本组89株金黄色葡萄球菌与ATCC29213阴性对照菌比较,22株能引起精子活力明显下降(P＜0.05);67株对精子活力的影响与ATCC29213阴性对照菌比较差异无统计学意义(P＞0.05).89株金黄色葡萄球菌对利奈唑胺、万古霉素和呋喃妥因全部敏感,对氟喹诺酮类(环丙沙星、左氧氟沙星)、红霉素、庆大霉素、四环素表现为较高的耐药性.本组89株金黄色葡萄球菌中,51株确定为MRSA,其中SCCmec Ⅰ型2株,SCCmecⅢ型12株,SCCmecⅣ型2株,SCCmecⅤ型25株,10株未能分型.PCR法检测金黄色葡萄球菌的14个毒力基因,发现22株携带hlg毒力基因、3株携带see毒力基因、1株携带sea毒力基因、1株携带seg毒力基因、1株携带sei毒力基因,1株同时携带hlg和sea基因,1株同时携带seg和sei基因. 结论 泌尿生殖道感染的不育症患者精液中分离的金黄色葡萄球菌能明显降低正常精子活力;本组金黄色葡萄球菌的主要毒力基因是γ-溶血素编码基因hlg,还有散在分布的肠毒素编码基因sea、see、seg和sei.

abstractsObjective To study the effect of Staphylococcus aureus from sperm of infertile male patients with genital urinary infection on spermatozoa,as well as the virulence genes spectrum of these Staphylococcus aureus.Methods A total of 89 strains of Staphylococcus aureus were isolated from infertility males'sperm between December 2009 and January 2011.The ATCC29213 strain of Staphylococcus aureus were considered as negative control.Computer assisted system:Sperm Class Analyzer V.3.2.0 was used to screen the influence of all strain on the robust semen according to the semen analysis method in the WHO (2010) guideline.The antimicrobial susceptibility test of all isolated strains was performed by agar dilution method including the common antibiotics,quinolones,sulfonamides and so on.The methicillin-resistant Staphylococcus aureus (MRSA) were determined by Cefoxitin disk diffusion and Polymerase Chain Reaction (PCR) for mecA gene.SCCmec genotypes were conformed via Multiplex-PCR and DNA sequencing,the virulence genes were also detected by PCR method.Results Twenty-two strains of Staphylococcus aureus could significiantly weaken the mobility of spermatozoahe than the ability in the negative control Staphylococcus aureus ATCC29213 (P＜0.05).The remainning 67 isolates could not inhibit the activity of sperm (P＞ 0.05).All the isolates were sensitive to the vancomycin,linezolid and furadantin.Instead,they displayed high-level antibiotics resistance to fluoroquinolones (Ciprofloxacin,levofloxacin),erythromycin,gentamicin and tetracycline.Among the 51 MRSA isolates,the SCCmec genotypes were shown as follows:SCCmec Ⅰ was indentified in 2 isolates,SCCmec Ⅲ in 12 isolates,SCCmec Ⅳ in 2 isolates,SCCmec Ⅴ in 25 isolates,the remaining ten were undefined genotyping.The putative 14 virulence determinants in the 89 isolates were detected by PCR,results were shown as follows:22 isolates of S.aureus had gene for hlg,3 isolates for see,1 isolate for sea,1 isolate for seg,1 isolate for sei,only 1 isolate had genes for hlg and sea simultaneously and 1 for seg and sei simultaneously.Conclusions The Staphylococcus aureus isolated from urogenital tract of infertile male patients with genitourinary infection could inhibit significantly the mobility of spermatozoa; In our research,the primary virulence determinate was hlg encoding γ-hemolysin,and the gene sea,see,seg and sei,which encoded enterotoxins were also scatteredly distributed in the Staphylococcus aureus.