性病门诊患者解脲脲原体分离株的分子分群及血清分型研究

目的了解性病门诊患者解脲脲原体（Uu）分离株的血清分型与分子分群的分布状况与相互关系。方法采用液体加固体的方法从临床标本中分离获得解脲脲原体临床菌株，纯化后采用PCR扩增Uu多条带基因进行分子分群，采用代谢抑制试验进行血清分型。结果从377例性病门诊患者中分离到Uu117例，阳性率31％。其中男性54例，阳性率46％；女性63例，阳性率54％。成功进行分子分群114例，其中生物1群47例，占41．2％；生物2群67例，占58．8％；进行血清分型60例，其中单个型别41例，占68．3％；多型别合并19例，占31．7％。出现频率高的型别为7型24例（29．63％），2型19例（23．46％），14型11例（13．58％）和4型7例（8．64％）。未见3、5、12、134种血清型别。16．7％Uu分离株（10／60）存在分型与分群结果不符情况。结论性病门诊患者Uu分离株中，生物2群所占比例多于生物1群，血清分型以7、2、14、4等型别常见，发现部分菌株分子分群与血清分型结果不符现象。     

荧光定量PCR检测解脲脲原体不同生物群的临床应用

目的 探讨解脲脲原体(Uu)的感染状态及生物分群.方法 采用荧光定量PCR 技术对皮肤科和泌尿外科患者157例尿道/宫颈分泌物进行Uu分群;分析Uu不同生物群与不同性别患者泌尿生殖道感染的临床症状和体征的相关性.结果 157例标本中96例Uu呈阳性,其中生物1群57例(阳性率59.38%),生物2群39例(阳性率40.62%),男性患者临床症状和体征的阳性率与Uu分群无关(P＞0.05),而女性生物1群感染者的临床症状和体征阳性率高于生物2群(P＜0.05).结论 我院患者Uu以生物1群为主,生物1群可能是女性Uu感染的主要病原体,Uu对男性致病性与分群无关.     

解脲脲原体对四环素的耐药性分析及其耐药机制研究

目的 了解解脲脲原体(Uu)对四环素的耐药性并分析其耐药机制.方法 采用支原体培养、鉴定、药敏试验一体化试剂盒进行药敏试验;用PCR基因扩增方法 对女性泌尿生殖道采集的200株Uu进行生物分群和耐药机制研究.结果 200株Uu中生物1群感染占70.0%(140/200),生物2群占22.0%(44/200).生物1群对四环素的耐药率为18.6%(26株);生物2群对四环素的耐药率为20.5%(9株).生物1群中耐四环素株的TetM基因阳性率88.5%(23/26);生物2群中9株耐四环素株中TetM基因阳性者8株.结论 女性泌尿生殖道分离的Uu大多数为生物1群.四环素可以作为治疗Uu感染的一线药物,但耐四环素的Uu携带TetM基因的比例很高,TetM基因的检出率与生物种群无关,仍应根据约敏试验结果 合理选用抗菌药物以延缓耐药株的产生.     

候选致病岛Uu146-Uu150在临床分离解脲脲原体中的检测分析

目的探讨候选致病岛（PAI）Uu146-Uu150在解脲脲原体（Uu）致病性中的意义。方法收集临床分离的Uu101株,运用多带抗原（MBA）．XLMycoplasmaIST2-iL剂盒对其进行生物分群及药敏检测．用特异性引物扩PAIUul46-Uu150。结果生物l群为杭州地区Uu的主要生物群别（占70．30％）;PAI检出率为11．88％;Fi8her检验结果显示,Uu生物1群PAI阳性率（7．04％）低于生物2群PAI阳性率（23．33％）,差异有统计学意义（P〈0．05）。结论PAIUu146-Uu150可能为一潜在的、与Uu生物2群相关的致病岛,但需要更多的流行病学调查和实验验证。     

宫颈解脲支原体的分群分型检测与致病性研究

目的 探讨有阴道脓性分泌物症状的患者与正常对照组感染解脲支原体(UU)的生物种群及血清分型的差异,从而为临床合理治疗提供依据.方法 用液体选择培养基初筛体检人群及门诊就诊患者319例,提取DNA,用PCR法进行分群分型.结果 231例有阴道脓性分泌物症状的患者中UU阳性率为34.6%(80例),对照组为29.5 %(26例),二者之间差异无统计学意义(P>0.05).观察组Ⅱ群阳性率47.5%明显高于正常对照组23.1%(P<0.05),而对照组Ⅰ群阳性率76.9%明显高于观察组52.5%(P<0.05).观察组较对照组1型感染率有增高趋势.3,6,14型Uu感染率相近.结论 有症状组与正常对照组Uu感染率相近,生物Ⅱ群与致病相关,临床应进一步进行Uu分群的检测.     

女性生殖道解脲支原体感染状况及其分群价值的研究

目的探讨女性生殖道解脲支原体感染状况以及分群对女性生殖健康的意义。方法选取拟诊断为阴道炎或者非淋菌性尿道炎患者220例作为病例组,选用健康查体者200例作为对照组,观察两组解脲支原体感染情况以及分群价值。结果 1病例组中检测到Uu阳性患者79例,阳性率为35.9%;对照组检测到Uu阳性为62例,阳性率为31.0%,两组受检者Uu阳性率差异无统计学意义(P0.05);2病例组中生物1群占72.2%,生物2群占24.1%,对照组生物1群占83.9%,生物2群占9.7%,两组患者生物1群和生物2群各自所占比例差异有统计学意义(P0.05)。感染后18h、24hUu2群明显高于Uu1群和对照组,差异有统计学意义(P0.05)。结论女性生殖道解脲支原体感染与亚群比例出现异常有关,生物2群具有毒性作用,临床上进行分群有利于对解脲支原体感染进行诊断。     

女性解脲脲原体感染与其男性伴侣解脲脲原体感染的关系

目的探讨女性解脲脲原体(Ureaplasma urealyticum,Uu)感染与其男性伴侣Uu感染的关系,为临床诊治Uu感染提供依据。方法对2010年2月至2011年3月就诊并排除其他病原体感染的有症状且Uu阳性女性334例,无症状但Uu阳性女性61例和同期男性伴侣395例,分别取宫颈管或阴道、尿道分泌物进行Uu实时荧光定量-聚合酶链反应(real-time fluorescence quantitative-Polymerase Chain Reaction,FQ-PCR)基因检测和PCR反向斑点杂交(PCR-RDB)基因分群检测。结果 395例男性伴侣中,203例(51.39%)经FQ-PCR检测为Uu阳性,其中,有症状和无症状女性的男性伴侣Uu阳性率分别为56.3%(188/334)和24.6%(15/61),差异有统计学意义(χ2=19.50,P0.01)。基因分群显示,有症状女性感染者生物1群、生物2群和混合群各占一定比例,但以生物1群占优势,而无症状女性感染者绝大部分以生物1群为主,占86.9%(53/61);有症状女性的男性伴侣中,有尿道炎症状者绝大部分(98.7%,147/149)为生物2群和混合群感染,无尿道炎症状者则绝大部分(94.9%,37/39)为生物1群感染;而无症状女性的男性伴侣均无尿道炎症状,感染主要以生物1群为主,占80.0%(12/15)。进一步分析发现,女性感染生物1群时其男性伴侣发生感染的比例较低,而女性感染生物2群和混合群时,其男性伴侣感染同类基因群的比例则高达98.6%(71/72)和73.0%(84/115)。结论无论女性感染生物1群还是生物2群,其男性伴侣感染的主要是生物2群。因此,在女性Uu感染者及其男性伴侣中进行Uu基因分群分析,不但可指导临床制定有针对性的治疗方案,而且对监测Uu的基因变化、追踪传染源、分析流行趋势有重要意义。     

溶脲脲原体对四环素类和大环内酯类抗生素的耐药性及其与血清型的关系

目的 :了解耐四环素类和大环内酯类溶脲脲原体 (Uu)的发生率和特性 ,探讨耐四环素Uu与血清型的关系。方法 :用微量肉汤稀释法对从性传播疾病 (STD)病人分离出来的 5 0株Uu菌株进行体外药敏试验和代谢抑制试验作血清学分型。结果 :Uu对四环素、多西环素、红霉素耐药率分别为 36 % (18/ 5 0 )、10 % (5 / 5 0 )和 5 0 % (2 5 / 5 0 ) ;且四环素、多西环素和红霉素存在交叉耐药。用代谢抑制试验对 10株四环素耐药和 10株四环素敏感的Uu菌株进行血清学分型 ,发现两者之间血清学分布无差异 (P >0 .0 5 ) ,但以Uu4型菌株常见。结论 :Uu对四环素、多西环素和红霉素存在交叉耐药 ,Uu菌株耐药性与血清学分布无明显差异 ,但以Uu4型菌株常见     

上海地区门诊性病患者支原体感染调查及其药敏结果分析

目的　了解上海地区性病门诊患者的解脲脲原体 (Uu)和人型支原体 (Mh)的感染及对罗红霉素(ROX) ,强力霉素 (DOX) ,氧氟沙星 (OFX)的药物敏感情况。方法　采用法国艾美生物技术公司的试剂盒检测340例患者。结果　 340例患者中支原体培养阳性 89例 ,阳性率为 2 6 .2 %。其中 ,单一Uu感染 4 1例 (12 .1% ) ;单一Mh感染 4例 (1.2 % ) ;Uu和Mh混合感染 4 4例 (12 .9% )。Uu感染 85株 ,其中耐DOX 8株 ,耐ROX 34株 ,耐OFX 39株 ;Mh感染 4 8株 ,其中耐DOX 5株 ,耐ROX 2 8株 ,耐OFX 2 8株。结论　上海地区性病患者支原体感染以Uu和Mh混合感染为主。Uu和Mh对ROX、OFX不敏感。单纯Mh感染可考虑使用DOX治疗。     

解脲脲原体改良培养基效果比较

目的比较解脲脲原体改良培养基、原Uu液相培养基及法国生物-梅里埃IST培养基(IST培养基)分离培养效果.方法 100例临床标本同时接种上述三种培养基,观察并记录解脲脲原体生长时间及阳性数,将三种培养基的阳性标本分别接种血平板以判断杂菌的生长情况.结果原Uu液相培养基、IST培养基、改良培养基对临床标本分离阳性率分别为37%、43%和45%,改良培养基较原Uu液相培养基分离率高(P＜0.05),生长速度快(P＜0.05),抑制杂菌能力强(P＜0.05).同进口IST培养基相比,差别没有显著性(P＞0.05).结论改良培养基同原Uu液相培养基相比具有分离阳性高、生长速度快及抑制杂菌能力强等优点,达到IST培养基同等效果.     

Genetic structure of Streptococcus pneumoniae isolated from children in a tertiary care university hospital, in Korea, 1995 to 2005

We investigated the genetic structure of Streptococcus pneumoniae isolated from Korean children prior to the use of heptavalent pneumococcal conjugate vaccine (PCV7). A total of 278 pneumococcal isolates were obtained from invasive (n = 77) and noninvasive (n = 201) specimens from children < 5 years of age that visited the Seoul National University Children's Hospital between 1995 and 2005; these samples were serotyped. Two hundred seventy-eight isolates were characterized by multilocus sequence typing, antibiotic susceptibility pattern testing with the E-test. The common serotypes were 19F (22.7%), 23F (19.1%), 19A (14.4%), 6B (11.2%), 6A (8.3%), 14 (5.4%), and 9V (4.3%); these 7 serotypes accounted for 85.3% of the 278 isolates. The isolates belonged to 23 serotypes and 81 sequence types (STs) (including 48 newly discovered STs). The eBURST analysis showed 7 clonal complexes (CCs). CC271 was the main CC of serogroup 19; CC81 and CC3115, serotype 6A; CC81 and CC880, serotype 23F; CC90, serotype 6B; CC166, serotype 9V; and CC554, serotype 14. Nonsusceptibility rates to penicillin and erythromycin among the invasive isolates were 9.1% and 94.8%, respectively. The resultant database may be useful for postvaccine surveillance.     

800例STD患者五种支原体感染状况的研究

本文应用nestedPCR (nPCR)技术对无锡地区 499例男性和 301例女性性传播疾病(STD)者的生殖道拭子进行了解脲脲原体 (Uu)、人型支原体 (Mh)、生殖支原体 (Mg)、肺炎支原体(Mpn)和发酵支原体 (Mf)等五种支原体联合检测。结果发现 ,男女性STD者Uu、Mh、Mg、Mpn、Mf检出率分别为 48.9%、19.8%、10.8%、7.4 %、1.0 %和 61.5%、34.2 %、7.6%、4.0 %和 0.%。无论男女性STD者最常见三种支原体感染模式为Uu单项阳性、Uu与Mh合并阳性和Mh单项阳性。并发现有三种或四种支原体合并感染的病例。表明我国男女性STD者中支原体感染情况较为严重。     

Comparison of genotypes of Streptococcus pneumoniae serotypes 6A and 6B before and after the introduction of PCV7 vaccination in Korea

We investigated changes of serotypes in Streptococcus pneumoniae and the characteristics of serotypes 6A and 6B in Korean hospitals after the introduction of the 7-valent pneumococcal conjugate vaccine (PCV7). PCV7 serotypes decreased significantly from 66.8% in 1996-2001 to 41.3% in 2008-2009 (P<0.001). However, serotype 6B did not decrease during these 2 periods (5.7% to 6.4%). Non-PCV7 serotypes that increased significantly were 3 (3.6% to 9.1%), 6A (3.1% to 8.5%), 13/28 (0 to 3.7%), 15 (2.2% to 6.7%), 16.36.37 (0 to 1.2%), 19A (1.2% to 9.7%), and 23A (0 to 1.5%). Multilocus sequence typing indicated that 9 STs were identified among 36 isolates of serotype 6A and 19 among 35 isolates of serotype 6B. No STs were found both in the 2 serotypes. The increase in serotype 6A was associated mainly with ST81, a Spain-23F clone genotype. The association of serotype 6A and ST81 is unique to Korea. Clones of serotype 6B isolates changed between the 2 periods. ST282, the main clone of serotype 6B in the PCV7 period, was found only in Korea. Our study demonstrates changing characteristics of pneumococcal serotypes 6A and 6B isolates in Korea and suggests selective evolution of certain clones in the era of PCV7 vaccination.     

Fluoroquinolone-Resistant Pneumococci: Dynamics of Serotypes and Clones in Spain in 2012 Compared with Those from 2002 and 2006

In Spain, rates of ciprofloxacin resistance in pneumococci were low during the last decade (2.6% in 2002 and 2.3% in 2006). In 2012, the rate remained at 2.3%, equivalent to 83 of 3,621 isolates. Of the 83 resistant isolates, 15 showed a low level (MIC of 4 to 8 μg/ml) and 68 a high level (MIC of 16 to 128 μg/ml) of ciprofloxacin resistance. Thirteen low-level-resistant isolates had single changes in ParC, one had a single ParE change, and one did not present any mutations. High-level-resistant isolates had GyrA changes plus additional ParC and/or ParE changes: 51, 15, and 2 isolates had 2, 3, or 4 mutations, respectively. Although 24 different serotypes were observed, 6 serotypes accounted for 51.8% of ciprofloxacin-resistant isolates: 8 (14.5%), 19A (10.8%), 11A (7.2%), 23A (7.2%), 15A (6.0%), and 6B (6.0%). A decrease in pneumococcal 7-valent conjugate vaccine (PCV7) serotypes was observed from 2006 (35.7%) to 2012 (16.9%), especially of serotype 14 (from 16.3% to 2.4%; P < 0.001). In comparison with findings in 2006, multidrug resistance was greater in 2012 (P = 0.296), mainly due to the increased presence and/or emergence of clonal complexes associated with non-PCV7 serotypes: CC63 expressing serotypes 8, 15A, and 19A; CC320 (with serotype 19A); and CC42 (with serotype 23A). Although rates of ciprofloxacin resistance remained low and stable throughout the last decade, changes in serotype and genotype distributions were observed in 2012, notably the expansion of a preexisting multidrug-resistant clone, CC63, and the emergence of the CC156 clone expressing serotype 11A.     

Changes of serotype and genotype in Streptococcus pneumoniae isolates from a Korean hospital in 2007

We investigated the change in clones and serotypes of Streptococcus pneumoniae isolates in a Korean tertiary-care hospital. Serotypes of S. pneumoniae isolates were determined by the capsular quellung method, and in vitro susceptibility testing was performed by broth microdilution method. Multilocus sequence typing was performed to determine the genotypes of the S. pneumoniae isolates. The erm(B) and mef(A) genes in erythromycin-resistant isolates were also detected using the duplex polymerase chain reaction method. During the 2 periods assayed (1998–2000 and 2007), 7-valent pneumococcal conjugate vaccine (PCV7) serotypes decreased significantly from 58.3% to 30.9% (P = 0.001). Especially, serotypes 19F and 23F decreased significantly from 31.7% to 8.5% (P < 0.0001) and from 20.0% to 7.4% (P = 0.021), respectively. In contrast to the other PCV7 serotypes, serotype 14 coupled with CC554 emerged in 2007, which may indicate no effect of PCV7 against serotype 14 isolates from Korea and the possibility of a different subtype. Of the non- PCV7 serotypes, serotype 19A increased from 8.3% to 14.9% (P = 0.227) and serotype 15 increased from 0% to 8.5% (P = 0.023). The increase of serotype 19A was due to the expansion of a preexisting clone with serotype 19A, ST320. However, S. pneumoniae isolates of serotype 15 showed diverse STs. Our data may provide helpful information in local vaccine serotype expansion or replacement in Korea.     

Changing trends in antimicrobial resistance and serotypes of Streptococcus pneumoniae isolates in Asian countries: an Asian Network for Surveillance of Resistant Pathogens (ANSORP) study

Antimicrobial resistance in Streptococcus pneumoniae remains a serious concern worldwide, particularly in Asian countries, despite the introduction of heptavalent pneumococcal conjugate vaccine (PCV7). The Asian Network for Surveillance of Resistant Pathogens (ANSORP) performed a prospective surveillance study of 2,184 S. pneumoniae isolates collected from patients with pneumococcal infections from 60 hospitals in 11 Asian countries from 2008 to 2009. Among nonmeningeal isolates, the prevalence rate of penicillin-nonsusceptible pneumococci (MIC, ≥ 4 μg/ml) was 4.6% and penicillin resistance (MIC, ≥ 8 μg/ml) was extremely rare (0.7%). Resistance to erythromycin was very prevalent in the region (72.7%); the highest rates were in China (96.4%), Taiwan (84.9%), and Vietnam (80.7%). Multidrug resistance (MDR) was observed in 59.3% of isolates from Asian countries. Major serotypes were 19F (23.5%), 23F (10.0%), 19A (8.2%), 14 (7.3%), and 6B (7.3%). Overall, 52.5% of isolates showed PCV7 serotypes, ranging from 16.1% in Philippines to 75.1% in Vietnam. Serotypes 19A (8.2%), 3 (6.2%), and 6A (4.2%) were the most prominent non-PCV7 serotypes in the Asian region. Among isolates with serotype 19A, 86.0% and 79.8% showed erythromycin resistance and MDR, respectively. The most remarkable findings about the epidemiology of S. pneumoniae in Asian countries after the introduction of PCV7 were the high prevalence of macrolide resistance and MDR and distinctive increases in serotype 19A.     

Serotype distribution and susceptibility of Streptococcus pneumoniae isolates from pleural fluid in Spain from 1997 to 2008

Trends in serotype incidence and susceptibility (1997 to 2008) of Spanish Streptococcus pneumoniae pleural isolates (n = 831) were explored. Penicillin (oral) nonsusceptibility rates and the incidence of 7-valent pneumococcal conjugate vaccine (PCV-7) serotypes showed decreasing trends (R(2) ≥ 0.600; P ≤ 0.002). The incidence of serotypes 1 and 19A showed increasing trends (R(2) ≥ 0.759; P < 0.001), with no trends for serotype 3. Serotypes 19A, 1, and 3 represented 85% of pediatric isolates in 2008. In serotype 19A, the penicillin nonsusceptibility rate was 82.4% in 2008, associated with amoxicillin and cefotaxime nonsusceptibility in 21.4% of isolates. Inclusion of these serotypes in new vaccines offers the broadest coverage.