孕晚期女性B群链球菌感染相关高危因素及其耐药性分析

目的 分析孕晚期女性B群链球菌（GBS）感染与孕产妇基础状态及伴随疾病之间的关系,探讨女性围产期GBS感染相关因素,分析GBS体外药物敏感性,指导GBS感染人群预防性抗菌药物的合理使用。方法 对2019年7月至2020年6月9 945名妊娠35～37周女性的阴道和肛周拭子进行GBS培养及分离鉴定,根据GBS检出情况分为GBS（+）组和GBS（-）组,对两组对象的年龄、生产史、流产史及疾病伴随情况进行单因素分析,采用二元Logistic回归分析GBS感染高危因素;对所有GBS（+）者行抗菌药物敏感性试验,分析GBS药物敏感情况。结果 兰州地区孕晚期女性生殖道GBS定植率为4.48%(446/9 945)。危险因素分析显示,年龄、生产史、流产史及妊娠合并糖尿病为GBS感染的高危因素（均P<0.05）。多因素二元logistic回归分析显示,年龄、生产史、流产史及妊娠合并糖尿病为影响GBS感染发生的独立危险因素（均P<0.05）。GBS体外药敏试验显示,GBS对氨苄西林、利奈唑胺、苄青霉素、奎奴普丁/达福普汀、万古霉素、替加环素、头孢噻肟及美罗培南均未出现耐药情况;对左氧氟沙星耐...     

B群链球菌的培养及优化研究

为了研究B群链球菌的生长和荚膜多糖的合成规律,采用摇瓶试验,探讨不同的液体培养基、培养基的pH值、葡萄糖的含量、生长因子、种子菌的接种量和溶氧等因素对B群链球菌生长的影响,优化发酵参数。并放大到10 L发酵罐中培养,菌浓度最高可达到20亿/mL;荚膜多糖的含量在培养的5 h达到652μg/mL。经过多次试验,建立了稳定的GBS发酵工艺。     

快速鉴定B群链球菌及检测其耐药性方法的建立和初步应用

目的建立快速鉴定B群链球菌(group B Streptococcus,GBS)的同时检测其对克林霉素、红霉素耐药基因的多重PCR方法,并初步应用。方法建立多重PCR方法,同时扩增以下4个基因位点:GBS编码CAMP因子的基因cfb、大环类脂类耐药基因erm B和mef A/E、克林霉素耐药基因lin B。对该方法的引物质量浓度、Taq酶量和退火温度进行优化,确定多重PCR最适反应体系及最低检测限;将该方法用于91株GBS临床菌株的鉴定及其耐药性的检测,并评价其与常规PCR方法结果的一致性。结果多重PCR的最适反应体系,总量25μL:多重PCR mix212.5μL、多重PCR mix1Taq酶0.23μL、lin B-F和lin B-R引物0.4μmol/L、mef A/E-F和mef A/E-R引物0.2μmol/L、erm B-F和erm B-R引物0.12μmol/L、cfb-F和cfb-R引物0.08μmol/L、DNA 10~100 ng、不足量补入无菌去离子水。扩增程序为:94℃预变性60 s;94℃变性30 s、53℃退火90 s、72℃延伸30 s,共30个循环;72℃延伸10 min。最低检测限为20 pg/μL。多重PCR方法与常规PCR方法检测91株GBS临床菌株,cfb基因结果一致率为100%,erm B、mef A/E和lin B基因的Kappa值分别为0.938、0.956和0.935。结论建立的以GBS cfb、erm B、mef A/E和lin B基因为检测位点的多重PCR方法,可在鉴定GBS的同时检测其对克林霉素、红霉素的耐药性。     

86株A群链球菌耐药性检测与分析

目的探讨A群链球菌对多种抗生素的耐药性,更好地指导临床用药。方法收集本院2005年度检出的39株和2006年度检出的47株A群链球菌的药物敏感试验结果,使用X^2检验比较A群链球菌耐药率。结果2006年度和上一年度相比A群链球菌对红霉素、克林霉素、阿齐霉素的耐药率增高显著（P〈0．05）,而对氯霉素、四环素的耐药性无明显改变（P〉0．05）,未发现耐万古霉素、青霉素、头孢吡肟、头孢噻肟的菌株。结论加强对A群链球菌耐药性检测及调查分析,对指导临床用药有重要意义。     

B群链球菌疫苗的研究

由于积极的调查和围产期抗生素的预防,目前分娩早期发作的GBS所致新生儿疾病流行呈可喜的下降趋势,有可能变成未来出现耐药GBS菌株的问题。GBS疫苗给这种条件致病菌的持久保护带来了最大的希望。本章将对GBS疫苗的研究进展做一综述。有效疫苗的研究进展是在具有医学、儿科学、多糖化学、免疫学、微生物学、遗传学,     

围产期生殖道菌群检测和B群链球菌筛查

目的 调查孕妇生殖道菌群情况,减少不良妊娠结果;筛查B群链球菌（GBS）定植状态,预防新生儿GBS病.方法 产前取阴道分泌物进行细菌培养,并对结果进行回顾性分析.结果 1464例孕妇阴道分泌物标本,872例乳酸杆菌和（或）少量其他菌种生长,占59.6%（872/1464）;592例乳酸杆菌未生长,检出“＋”以上生长菌株820株,其中194例有2种菌生长,34例有3种菌生长,占40.4%（592/1464）.共检出B群链球菌68株,检出率为4.6%（68/1464）.结论 围产期生殖道菌群种类复杂,耐药率高,妇幼保健工作人员应加强检测,采取切实可行的适宜技术,提供进一步的干预措施和保健服务.     

环介导等温扩增快速检测B群链球菌的临床应用

目的建立采用环介导等温扩增（LAMP）技术从临床标本中快速检测B群链球菌的方法。方法根据美国国家生物信息中心上提交的B群链球菌的cfb基因序列（登陆号X72754）设计特异LAMP引物,以热裂解法提取标本中细菌的DNA,然后以LAMP技术扩增cfb基因来鉴定其是否为B群链球菌。在采用LAMP技术检测B群链球菌的同时,以选择性培养法和PCR技术平行检测相同标本,并将3种方法的检测结果比较。结果在176例阴道拭子和176例直肠拭子中,选择性培养法检测到的B群链球菌例数为49和61、LAMP法检测到的B群链球菌例数为49和59,PCR法检测到的B群链球菌例数为48和58。以选择性培养法为金标准,LAMP法检测B群链球菌的灵敏度和特异度分别为96．7％和100％,PCR法检测B群链球菌的灵敏度和特异度分别为95．1％和100％。LAMP法检测B群链球菌的时间为2h左右,模拟菌株的检测结果显示该方法的最低检测限为10^2CFU／mL。结论该方法灵敏度高,特异性强,操作方便简单,适合从临床标本中快速检测B群链球菌。     

孕妇产道B群链球菌的调查报告

由B群链球菌引起的脑膜炎、肺炎及败血症严重威胁新生儿的生命。其传染源为产妇产道。本文采集143例临产孕妇消毒前的产道拭子及32例新生儿口咽部拭子,经增菌后培养在羊血平板,用生化方法及SPA血清分群法对疑似链球菌体鉴定。孕妇标本B群链球菌的检出率为11.2%,其他链球菌有D群、G群及草绿色链球菌。新生儿标本几乎都无细菌生长。纲菌鉴定方法以SPA法优于生化法。笔者认为由于B群链球菌在临床上的重要性,该菌的分离培养及鉴定应在实验室受到重视。     

围产期B族链球菌感染及预防的研究进展

B群链球菌可以引发产褥期败血症、新生儿肺炎和脑膜炎等感染,围产期B群链球菌感染和预防研究有着重要的价值。本研究就围产期女性B群链球菌感染、检测方法及预防相关研究进展作一综述,目的在于提高围产学界对该类微生物的认识。     

儿科痰培养肺炎链球菌耐药性分析

目的了解厦门市妇幼保健院儿科患者下呼吸道感染肺炎链球菌的耐药情况。方法对厦门市妇幼保健院2007年9月至2009年12月从儿科分离的95株肺炎链球菌用K-B纸片法检测其对青霉素等5种抗菌药物的敏感度,对耐苯唑西林的菌株用E-test法检测青霉素和头孢曲松的MIC值。结果95株肺炎链球菌对红霉素、克林霉素高度耐药,耐药率分别为97.9%、96.8%,对左氧氟沙星几乎敏感,未检出耐万古霉素菌株。E-test法测试青霉素不敏感率为58.9%（56株）,头孢曲松敏感率为93.7%（89株）。ermB因介导的红霉素耐药菌株92株,占98.9%,mefE基因介导的红霉素耐药仅1株（1.1%）。结论厦门市妇幼保健院分离的肺炎链球菌耐药严重,PNSP检出率较高（58.9%）,对红霉素、克林霉素高度耐药,临床应尽量减少此类药物的经验性用药,依据药敏结果选择抗菌药物进行治疗。     

Drug resistance of Enterococcus faecium clinical isolates and the conjugative transfer of gentamicin and erythromycin resistance traits

Drug resistance and the transferability of resistance were examined in 218 Enterococcus faecium clinical isolates obtained from in-patients of a Japanese university hospital between 1990 and 1999. One hundred and sixty one isolates (73.9%) were drug-resistant and 127 (58.2%) isolates were resistant to two or more drugs. Vancomycin resistant E. faecium (VRE) was not isolated. The transferability of drug-resistance to an E. faecium strain was examined by broth or filter mating. Six (12.5%) of the 48 gentamicin resistance traits, and fifty (50%) of the 101 erythromycin resistance traits were transferred by filter mating. The gentamicin resistance traits of five isolates and the erythromycin resistance traits of four isolates were transferred to the recipient strains by both broth mating and filter mating at a frequency of about 10(-6) and 10(-5) per donor cell, respectively. The five gentamicin resistant strains were shown to harbor pMG1-like plasmids on the basis of their Southern hybridization with pMG1 (65.1 kbp, Gm(r)), which transfers efficiently between enterococci by broth mating. Each of the four erythromycin resistant transconjugants obtained by broth mating harbored a large conjugative plasmid (more than 100 kbp). The plasmids showed no homology with well-characterized enterococcal conjugative plasmids such as pAD1, pPD1, pAM(beta)1, pIP501 and pMG1 by Southern hybridization. Of the erythromycin resistance traits that transferred only by filter mating, it was found that the erythromycin resistance trait was conferred by a 47-kbp transposable element that transferred from the chromosome of the donor strain to different sites within the pheromone responsive plasmid pAD1 (60 kbp) of the recipient strain, suggesting that the erythromycin resistance trait was encoded on a conjugative transposon, which was named Tn950.     

围产期孕妇泌尿生殖道无乳链球菌耐药性分析及临床意义

目的 了解围产期孕妇泌尿生殖道无乳链球菌感染及耐药分布为临床合理用药提供依据,并提出预防措施,以引起临床重视.方法 对门诊、住院围产期孕妇送检的泌尿生殖道标本进行培养鉴定,并对分离的无乳链球菌进行药敏试验,并用WHONET 5.5软件分析.结果 260株无乳链球菌药敏结果显示,对万古霉素、利奈唑胺、青霉素和头孢曲松的耐药率最低,对红霉素、克林霉素、左氧氟沙星的耐药率较高,分别为61.5％、57％和49.3％.结论 临床医生应重视对围产期孕妇无乳链球菌的检测,实验室应准确报告无乳链球菌药敏结果,对临床合理用药治疗无乳链球菌感染具有重要指导意义并可预防其危害性.     

Identification of two plasmids determining resistance to tetracycline and to erythromycin in group D streptococcus

Summary Two circular DNA plasmids designated Tet and Ero, distinguishable by their molecular weights (64.5 and 17.6×10⁶ respectively) and their guanine plus cytosine (G+C) content (36.5 and 34.5% respectively) were identified in Streptococcus faecalis strain KR, resistant to tetracycline and erythromycin. Loss of resistance to tetracycline, to erythromycin and to both antibiotics simultaneously in independently isolated cured derivatives corresponds to the loss of the Tet, Ero, and of both plasmids respectively. In the singly resistant strains we estimated that 3 and 6 copies of the Tet and Ero plasmids respectively are present per chromosomal genome equivalent.Abbreviations form I supercoiled circular DNA - form II relaxed circular DNA - form III linear double-strand DNA - Tet tetracycline - Ero erythromycin - R resistant - S sensitive     

In vitro compared activity of telithromycin and azithromycin against northwest Italian isolates of Streptococcus pyogenes and Streptococcus pneumoniae with different erythromycin susceptibility

Aims: This study compared the in vitro activity of telithromycin with that of azithromycin against 438 Streptococcus pyogenes and 198 Streptococcus pneumoniae, isolated over the period 2005–2007 from specimens of different human origin obtained in three Piemonte Region’s hospitals. Methods and Results: The determination of antimicrobial activity was evaluated by the microdilution broth method and the erythromycin‐resistant (Ery‐R) phenotypes by the triple‐disc test. Exactly 78·8% of S. pyogenes and 69·2% of S. pneumoniae were erythromycin‐susceptible (Ery‐S). Concerning S. pyogenes, telithromycin was active against M and inducible MLS_B, subtype‐C, phenotypes but not against constitutive MLS_B strains. Telithromycin acted well against all S. pneumoniae, irrespective of their mechanism of macrolide‐resistance. On the contrary, the Ery‐R isolates, both S. pyogenes and S. pneumoniae, were resistant to azithromycin. Conclusions: Our results indicate that macrolide resistance in streptococci still persist in northwest Italy (21·2% of S. pyogenes and 30·8% of S. pneumoniae) and that telithromycin is confirmed as being extremely active even against recent clinical Ery‐R streptococcal isolates. Significance and Impact of the Study: The present study emphasizes that an active surveillance of the phenotype distribution and antibacterial resistance in streptococci is essential in guiding the effective use of empirical treatment option for streptococcal infections, also at regional level.     

Genetics of high level penicillin resistance in clinical isolates of Streptococcus pneumoniae

Abstract Mosaic penicillin-binding proteins (PBP) 1A, 2X and 2B genes were cloned from four clinical isolates of Streptococcus pneumoniae with levels of susceptibility to penicillin ranging from 1.5 to 16 μg benzylpenicillin ml⁻¹. In each instance it was possible to transform either the penicillin-sensitive laboratory strain R6 or a sensitive clinical isolate 110K/70 to the full level of penicillin resistance with these three penicillin-binding proteins alone. Until now it has not been possible to clearly determine whether alterations to PBP1A, 2X and 2B alone were sufficient to attain high level penicillin resistance.     

长沙地区临床分离金黄色葡萄球菌的耐药监测

目的了解长沙地区临床分离金黄色葡萄球菌（以下简称金葡菌）对常用抗菌药物的耐药现状,探讨金黄色葡萄球菌对甲氧西林的耐药水平。方法收集长沙地区11家医院2009年11月至2010年11月临床分离的非重复金葡菌279株,应用Vitek-2全自动微生物分析系统进行鉴定,K-B法检测金葡菌对24种药物的敏感性,产色头孢菌素试验检测β-内酰胺酶以及D试验检测诱导型克林霉素耐药。应用头孢西丁和苯唑西林纸片扩散法筛查耐甲氧西林的金葡菌（MRSA）,琼脂稀释法检测头孢西丁和苯唑西林的最低抑菌浓度（MIC）。结果在被检测的24种药物中,敏感率〉50%的药物为9种,未发现对万古霉素、替考拉宁和利奈唑胺耐药菌株;耐药率〉50%的抗菌药物有11种,其中以青霉素和氨苄西林的耐药率最高（均为97.1%）。MRSA的分离率达54.5%,且对常用的16种抗菌药物的耐药率均显著高于甲氧西林敏感金黄色葡萄球菌（MSSA）。279株金葡菌中,β-内酰胺酶阳性250株（89.6%）;红霉素耐药而克林霉素敏感或中介的30株中,D试验阳性22株（73.3%）。苯唑西林（OXA）和头孢西丁（FOX）MIC范围分别为0.125～〉256μg/mL和2～〉256μg/mL,苯唑西林的MIC50和MIC90分别为128μg/mL和256μg/mL,头孢西丁的MIC50和MIC90分别为64μg/mL和256μg/mL。结论长沙地区临床分离金葡菌对常用抗菌药物呈多重耐药;MRSA不仅分离率高,而且对甲氧西林呈高水平耐药。     

A mutant of group A streptococcus resistant to high levels of kanamycin exhibits characteristics of penicillin tolerance in vitro

Abstract In studies of penicillin tolerance in group A streptococci, we observed that a mutant of group A streptococcus isolated for high-level resistance to kanamycin exhibited several characteristics of penicillin tolerance: significant disparity between the penicillin MIC and MBC, delayed killing by penicillin concentrations of 16 times the MIC, and survival in areas of superinhibitory penicillin concentrations when strains were transferred from a penicillin-gradient plate to a penicillin-free replicate plate. In contrast, the parent strain of group A streptococcus and its mutant isolated for high-level resistance to streptomycin were nontolerant for penicillin. Moreover, a clinical isolate of group A streptococcus possessing high-level resistance to kanamycin was also found to be tolerant to penicillin. These findings suggest that genetic mechanisms responsible for high-level resistance to kanamycin may be related with the expression of penicillin tolerance of group A streptococci in vitro.     

肺炎克雷伯菌临床分离株的庆大霉素耐药性和细菌铁载体的作用

目的 研究临床分离的肺炎克雷伯菌对氨基糖苷类抗生素庆大霉素的耐药性与其产铁载体的关系。方法 采用K-B纸片法和肉汤稀释法确定70株临床分离的肺炎克雷伯菌对庆大霉素的药物敏感性;CAS琼脂实验检测肺炎克雷伯菌是否产铁载体;紫外可见分光光度法确定细菌产铁载体的量,根据中位数法将70株临床分离菌分为铁载体高产组（35株）和低产组（35株）;应用SPSS统计学软件分析抗生素耐药性与其产铁载体是否相关。结果 药物敏感性试验检测出菌株对庆大霉素的耐药率为50.00%（35/70）;铁载体检测实验确定70株肺炎克雷伯菌均产生铁载体,肺炎克雷伯菌对庆大霉素的耐药性与铁载体产量呈正相关关系（r=0.3154,P<0.05）,对庆大霉素耐药菌株铁载体产量明显高于敏感菌株（t=3.1650,P<0.05）,且铁载体高产组耐药率及lgMIC值明显高于低产组（χ2=9.6570,t=3.1360,P<0.05）。结论 70株临床分离的肺炎克雷伯菌均产生铁载体,铁载体可能参与肺炎克雷伯菌对庆大霉素的耐药,干扰庆大霉素的抑菌或杀菌过程。