207例男性尿道炎患者的解脲脲原体药敏分析

目的 了解广州地区解脲脲原体（Uu）的耐药性,更合理选择抗生素。方法 采用支原体药敏检测试剂盒和支原体固体培养基进行Uu培养、鉴定和药敏检测。结果 Uu阳性率为21.7%,强力霉素、美满霉素、环丙沙星、氧氟沙星、诺氟沙星、罗红霉素、阿奇霉素、克拉霉素、交沙霉素和壮观霉索耐药率依次为3.4%、1.4%、53.6%、29%、1.4%、2.4%、0.05%、24.6%和13.5%。结论 广州地区对于Uu引起的非淋菌性尿道炎（NGU）,首选药物是以强力霉素、美满霉素、诺氟沙星、罗红霉素、阿奇霉素和克拉霉素为主,开展体外药敏试验,根据试验结果合理选择治疗药物。     

1796例泌尿生殖道支原体培养及药敏结果分析

目的分析新乡地区1796例泌尿生殖道标本的支原体培养及抗生素药物敏感试验,指导临床合理使用抗生素。方法支原体培养鉴定、药敏试剂盒购自贝瑞特公司。同时检测解脲支原体和人型支原体,并进行10种抗生素的药敏试验。数据统计采用χ^2检验。结果支原体培养1596例标本中,541例解脲脲原体（Uu）阳性,阳性率为33.9%;39例人型支原体（Mh）阳性,阳性率为2.4%;解脲脲原体、人型支原体均阳性76例,阳性率为4.8%。200例正常对照组27例阳性,阳性率为13.5%。药敏结果显示,强力霉素、克拉霉素和美满霉素的抗菌活性较强。结论支原体感染以Uu为主,Uu＋Mh次之,正常人泌尿生殖道支原体阳性率为13.5%。只有某些血清型感染且在达到一定浓度以上同时宿主处于多种病原体感染或免疫机制紊乱时,Uu才能致病。避免滥用抗生素引起生殖道菌群失调。临床上治疗Uu感染、Mh感染或Uu＋Mh混合感染时,建议选用强力霉素、克拉霉素和美满霉素。     

解脲脲原体耐药性研究进展

解脲脲原体是条件致病菌。目前对其耐药性的研究主要包括对喹诺酮类、大环内酯类和四环素类3种抗菌药物相关耐药突变位点的检测,以及生物膜对病原体相关药物敏感性的影响,研究方法和检验手段仍较为传统、局限,研究方案也仅停留在对前人实验的重复。近年来,有学者将多位点序列分型技术用于解脲脲原体耐药序列类型的研究。在完善耐药机制研究的基础上,如何实现对耐药株的快速鉴定,从而指导抗菌药物的合理选择等仍需进一步研究。     

棋盘稀释法在解脲脲原体药敏试验中的应用

解脲脲原体是引起泌尿生殖道感染的重要病原体。目前使用的药敏试验方法即药敏卡法没有考虑样本中脲原体的实际含量而存在局限性。本研究将棋盘稀释法应用于解脲脲原体的药敏试验,可获得脲原体的准确耐药情况,有助于临床治疗中抗生素的正确选择;同时,有利于控制耐药菌株的产生和在人群中的扩散。     

大学生解脲脲原体和人型支原体正常携带状况研究

目的探讨解脲脲原体(Uu)和人型支原体(Mh)在大学生中的正常携带状况.方法采用培养法对314名未婚统招大学生和232名已婚成教大学生进行解脲脲原体和人型支原体的检测.结果未婚大学生和已婚大学生泌尿生殖道支原体检出率分别为12.10%和22.41%,后者明显高于前者(x2=10.31,P＜0.005);已婚男、女大学生泌尿生殖道支原体检出率(分别为17.44%和27.78%)明显高于未婚男、女大学生检出率(分别为8.33%和16.44%,x2=5.84、4.77P＜0.05).未婚男、女大学生之间泌尿生殖道支原体检出率差异有显著性(y2=4.82,P＜0.05),而已婚男、女大学生之间泌尿生殖道支原体检出率差异无显著性(x2=3.34,P＞0.05).已婚和未婚大学生泌尿生殖道支原体检出者中均以同时携带Uu和Mh较为常见.结论在校大学生中也有一部分人正常携带Uu和/或Mh,从他(她)们体内检出Uu和/或Mh一般不代表疾病状态.     

对不同浓度解脲脲原体的药敏结果分析

目的 了解不同浓度解脲脲原体＜10^4CFU/ml和≥10^4CFU/ml耐药率情况.方法 采用珠海黑马生物工程有限公司集分离、培养、计数、药敏于一体的试剂盒,按说明进行操作.结果 交沙霉素、美满霉素、强力霉素的总耐药率较低,左旋氧氟沙星、司帕沙星、罗红霉素、阿奇霉素、克林霉素对不同浓度解脲脲原体的耐药率差异有高度显著性.结论 交沙霉素、美满霉素、强力霉素可作为治疗解脲脲原体的首选药物,克林霉素总耐药率已高达57.7%（164/284）,不适宜用于解脲脲原体的治疗.珠海黑马产品质量优于其它同类产品.     

慢性阴道炎患者生殖道解脲脲原体和沙眼衣原体感染分析

目的通过荧光定量聚合酶链反应(PCR)技术检测慢性阴道炎患者阴道内解脲脲原体(Uu)和沙眼衣原体(Ct)感染状况,用以指导临床正确治疗。方法应用荧光定量聚合酶链反应对380例慢性阴道炎患者的阴道分泌物进行解脲脲原体和沙眼衣原体PCR检测。结果解脲脲原体阳性率为42.9%,其阳性标本的平均拷贝数为3.4×105copies/ml,沙眼衣原体阳性率为16.6%,其阳性标本的平均拷贝数为5.8×104copies/ml,解脲脲原体和沙眼衣原体合并感染的阳性率为12.6%。结论PCR技术具有简便、快捷、准确的优点,是目前快速诊断慢性阴道炎患者Uu、Ct感染状况的可靠诊断方法之一。     

2194例男性泌尿生殖道支原体检测及药敏分析

目的研究男性泌尿生殖系统炎症患者解脲脲原体(Uu)和人型支原体(Mh)感染及对抗菌药物耐药情况,以了解本地区男性支原体的感染流行状况并指导临床合理选择使用抗菌药物。方法采用支原体鉴定药敏试剂盒,对男性生殖道感染者进行支原体培养及药敏试验。结果 2 194份标本中检出支原体636例,阳性率为28.96%,其中解脲脲原体阳性率为22.59%,人型支原体阳性率为0.96%,解脲脲原体和人型支原体混合感染阳性率为5.42%。支原体对抗菌药物药敏结果显示,抗菌活性较高的是多西环素、普拉霉素、四环素和交沙霉素。结论Uu是男性泌尿生殖系统支原体感染的主要病原体。临床治疗支原体感染时应尽量根据药敏结果选择敏感药物。普拉霉素、多西环素、四环素和交沙霉素可作为本地区非淋菌尿道炎经验用药的首选药物,避免使用喹诺酮类药物。     

2007-2011年杭州市萧山区女性支原体感染及耐药性分析

目的 调查女性泌尿生殖道炎症解脲脲原体(Uu)和人型支原体(Mh)感染及其对抗菌药物耐药性,以了解本地区女性支原体感染状况并指导临床合理使用抗菌药物.方法 采用生物梅里埃Mycoplasma IST支原体鉴定、药敏试剂盒,对女性生殖道感染患者进行支原体培养及药敏试验,所有数据采用WHONET 5.6软件进行回顾性分析.结果 14 894份标本中检出支原体8319例,阳性率55.9％,其中解脲脲原体6 316例(42.4％),人型支原体282例(1.9％),解脲脲原体和人型支原体混合感染1 721例(11.6％).支原体对喹诺酮类耐药率较高(≥39.8％),对多西环素、普拉霉素、四环素和交沙霉素耐药率较低(≤7.0％).结论 Uu是女性泌尿生殖道支原体感染的主要病原体;多西环素、四环素和交沙霉素可作为本地区非淋病性尿道炎(NGU)的首选药物,环丙沙星耐药率出现明显的上升趋势,经验用药避免使用喹诺酮类药物,治疗时应根据药敏结果合理选用抗生素以防止耐药菌株的增加和播散.     

运用PCR 扩增技术检测解脲脲支原体生物群的临床应用

目的:探讨运用酶链聚合反应(PCR)技术检测泌尿生殖道解脲脲原体(Uu)的生物群,分析Uu生物群与临床症状的相关性。方法:以支原体16S rRNA保守区域基因为扩增靶序列设计引物,采用PCR方法扩增Uu 16S rRNA基因检测125例临床标本,并将检测结果与临床症状进行相关性分析。结果:PCR法检出Uu阳性率44.8%,其中35例Uu生物1群阳性,其中有16例有症状;27例Uu生物2群阳性,其中有18例有症状。Uu生物1群感染与症状的相关性无统计学差异(P>0.05),而Uu生物2群感染与症状相关性有统计学差异(P<0.05)。结论:PCR检测Uu 16S rRNA基因可用于Uu生物群的检测,Uu生物2群可能是非淋菌性尿道炎(NGU)的一个致病菌,而Uu生物1群与NGU无明显相关性。     

Assessing and analysing contamination of a dairy products processing plant by Staphylococcus aureus using antibiotic resistance and PFGE

A dairy product processing plant was studied for 2.5 years to examine contamination with Staphylococcus aureus and try to correlate the source of contamination. Cultures were submitted to an antibiotic susceptibility test (AST) and characterised by Pulsed-field Gel Electrophoresis (PFGE) analysis. Results showed that 35.2% (19/51) of food handlers were asymptomatic carriers of S. aureus, and that 90.4% (19/21) of raw milk sampled was contaminated. Staphylococcus aureus was isolated from only 10 samples among more than 3200 investigated dairy products. No S. aureus contamination was found on machinery. The AST analysis demonstrated sensitivity of tested S. aureus to oxacillin, cephalothin, vancomycin, gentamicin, and sulfamethoxazole/trimethoprim. AST analysis generated eight different phenotypic profiles, but did not allow us to identify the source of contamination in seven of ten final products. PFGE analysis proved to be a sensitive method as it generated 42 different DNA banding profiles among the 48 S. aureus investigated, demonstrating a lack of predominance of endemic strains in the plant, contrary to suggestions raised by antibiotic resistance typing. Based on PFGE genotyping, S. aureus strains isolated from four contaminated final products were similar to four S. aureus isolated from raw milk. Five final products contained S. aureus different from all other strains collected, and one showed similarity to a strain isolated from a food handler. These results suggest contamination by raw milk as the main source of contamination of the final dairy products.     

水产病原菌抗菌药物敏感试验标准化探讨

病原菌药物敏感试验是临床微生物学研究的重要手段,相比人类和兽医临床,国内外水产养殖中细菌性病原的药敏试验还未有公认的标准化参考方法。概括近几年国外对水产病原菌药敏试验标准化方法建立的研究进展,探讨了试验中关键因子的影响,指出我国水产药敏试验中存在的不足及对未来发展提出展望。     

Creation of an experimental rearing environment for microbiome animal research using an individually ventilated cage system and bioBUBBLE enclosure

To avoid microbial contamination risk, vinyl film isolators are generally used in animal microbiome experiments involving germ-free (GF) mice and/or gnotobiotic (GB) mice. However, it can take several months to gain expertise in operating the isolator competently. Furthermore, sterilization and sterility testing, which are essential for isolator preparation, can take more than 20 days. Hence, we built an experimental rearing environment that combines an individual ventilation cage system and a bioBUBBLE clean room enclosure to easily set up an experimental animal microbiome environment for animal facilities. In this work, a three-step evaluation was conducted. First, we examined whether GF mice can be maintained in this rearing environment without bacterial contamination. Next, we examined whether GF and GB mice can be maintained without cross-contamination in one individual ventilation cage rack. Finally, we tested whether GF mice can be maintained in a biological safety cabinet controlled by negative pressure. In our series of experiments, no microbial contamination occurred over more than 3 months. These results indicated that our rearing system that combines the individual ventilation cage and bioBUBBLE systems can be used not only for experiments with GF mice but also for Biosafety Level 2 experiments that handle bacteria. Our system can mitigate various disadvantages of using vinyl film isolators. In conclusion, we established an experimental method with improved working time and efficiency compared with those of the previous vinyl isolator method.     

Identification and Antimicrobial Susceptibility Testing of <Emphasis Type="Italic">Staphylococcus vitulinus</Emphasis> by the BD Phoenix Automated Microbiology System

This study evaluated the performance of the BD Phoenix system for the identification (ID) and antimicrobial susceptibility testing (AST) of Staphylococcus vitulinus. Of the 10 S. vitulinus isolates included in the study, 2 were obtained from the Czech Collection of Microorganisms, 5 from the environment, 2 from human clinical samples, and 1 from an animal source. The results of conventional biochemical and molecular tests were used for the reference method for ID, while antimicrobial susceptibility testing performed in accordance with Clinical and Laboratory Standards Institute recommendations and PCR for the mecA gene were the reference for AST. Three isolates were incorrectly identified by the BD Phoenix system; one of these was incorrectly identified to the genus level, and two to the species level. The results of AST by the BD Phoenix system were in agreement with those by the reference method used. While the results of susceptibility testing compared favorably, the 70% accuracy of the Phoenix system for identification of this unusual staphylococcal species was not fully satisfactory.     

Preparation of a Blood Culture Pellet for Rapid Bacterial Identification and Antibiotic Susceptibility Testing

Bloodstream infections and sepsis are a major cause of morbidity and mortality. The successful outcome of patients suffering from bacteremia depends on a rapid identification of the infectious agent to guide optimal antibiotic treatment. The analysis of Gram stains from positive blood culture can be rapidly conducted and already significantly impact the antibiotic regimen. However, the accurate identification of the infectious agent is still required to establish the optimal targeted treatment. We present here a simple and fast bacterial pellet preparation from a positive blood culture that can be used as a sample for several essential downstream applications such as identification by MALDI-TOF MS, antibiotic susceptibility testing (AST) by disc diffusion assay or automated AST systems and by automated PCR-based diagnostic testing. The performance of these different identification and AST systems applied directly on the blood culture bacterial pellets is very similar to the performance normally obtained from isolated colonies grown on agar plates. Compared to conventional approaches, the rapid acquisition of a bacterial pellet significantly reduces the time to report both identification and AST. Thus, following blood culture positivity, identification by MALDI-TOF can be reported within less than 1 hr whereas results of AST by automated AST systems or disc diffusion assays within 8 to 18 hr, respectively. Similarly, the results of a rapid PCR-based assay can be communicated to the clinicians less than 2 hr following the report of a bacteremia. Together, these results demonstrate that the rapid preparation of a blood culture bacterial pellet has a significant impact on the identification and AST turnaround time and thus on the successful outcome of patients suffering from bloodstream infections.     

Frequent major errors in antimicrobial susceptibility testing of bacterial strains distributed under the Deutsches Krebsforschungszentrum Quality Assurance Program

The Quality Assurance Program (QAP) of the Deutsches Krebsforschungszentrum (DKFZ) was a proficiency testing system developed to service the laboratory animal discipline. The QAP comprised the distribution of bacterial strains from various species of animals for identification to species level and antibiotic susceptibility testing (AST). Identification capabilities were below acceptable standards. This study evaluated AST results using the DKFZ compilations of test results for all bacterial strains showing the number of participants reporting the strain as resistant (R), sensitive (S) or intermediate susceptible (I) to each antibiotic substance used. Due to lack of information about methods used, it was assumed that what the majority of the participants reported (R or S) was the correct test result and that an opposite result was a major error (ME). MEs occurred in 1375 of 14,258 (9.7%) of test results and ME% ranged from 0% to 23.2% per bacterial group-agent group combination. Considerable variation in MEs was found within groups of bacteria and within groups of agents. In addition to poor performance in proper species classification, the quality of AST in laboratory animal diagnostic laboratories seems far below standards considered acceptable in human diagnostic microbiology.     

癌症靶向基因-病毒ZD55-XAF1抗肝癌移植瘤的生长及其安全性研究

目的:探讨溶瘤腺病毒(ZD55-gene)作为载体携带外源抗癌基因(XAF1)抗肝癌移植瘤的生长及其安全性。方法:抽提溶瘤腺病毒ZD55-XAF1的基因组DNA,PCR扩增鉴定病毒;细菌平板培养和支原体检测试剂盒检测细胞有无细菌、支原体污染;通过荷瘤小鼠动物实验,观察溶瘤腺病毒ZD55-XAF1对肝癌移植瘤生长的抑制、小鼠的临床反应指标、血清肝毒性指标、各脏器组织中的病毒残留分布及病理切片观察。结果:细胞培养过程无细菌和支原体污染;较对照组,受试小鼠血清肝酶AST活性上升(P0.05),而ALT和ALP活性基本无变化(P0.05);PCR检测各脏器均有病毒基因组DNA存在;HE染色显示受试小鼠各脏器具有不同程度的损伤,病毒处理对肿瘤细胞具有明显的杀伤效果,而受试小鼠的临床反应并无明显异常。结论:溶瘤腺病毒ZD55-XAF1能够抑制肿瘤生长,杀死肿瘤细胞,对小鼠血清肝酶活性影响较小而对各脏器有不同程度的轻微损伤,作为癌症基因治疗载体有潜在的应用价值但其安全性还有待提高。     

Early effects of tribromoethanol, ketamine/xylazine, pentobarbitol, and isoflurane anesthesia on hepatic and lymphoid tissue in ICR mice

We investigated the effects of various anesthetic agents on hepatic and splenic injury in mice. Three and six hours after intraperitoneal injection of TBE, intramuscular injection of ketamine/xylazine combination (K/X), intraperitoneal injection of pentobarbital (PB), and inhalation of isoflurane (IF), or intraperitoneal and intramuscular injection of control saline, mice were exsanguinated and serum was obtained for measurement of hepatic aspartate transaminase (AST), alanine transaminase (ALT) and gamma-glutamyltransferase (GGT). The spleen and liver also were obtained, and sections were examined by use of routine light microscopy for pathologic changes and for apoptosis, as determined by use of the in situ terminal deoxynucleotidyl transferase-mediated dUPT nick-end-labeling (TUNEL) histochemical analysis. Three hours after TBE or K/X administration, AST activity increased three- to fourfold above that in untreated and saline-injected control animals, and remained high at six hours. Administration of PB did not effect AST activity at three hours, but there was a significant increase at six hours. Activity of ALT was non-significantly increased three hours after TBE and K/X, but not PB administration. Administration of IF had no effect on hepatic enzyme activities, and GGT was not increased after administration of any of the agents. Markedly increased apoptosis was observed in splenic follicles and in hepatic Kupffer and endothelial cells at three hours after TBE and K/X administration, but apoptosis decreased to control levels by six hours. Increased apoptosis was not observed after IF administration. Administration of TBE and K/X causes injury to lymphocytes and to hepatic Kupffer and endothelial cells within three hours, and PB administration induces changes within six hours. Thus, use of these anesthetic agents should be avoided when experiments are being designed to test short-term effects of an experimental intervention on the spleen and possibly on all lymphoid tissues. In addition, they also should be avoided in experiments testing effects on hepatic tissue.     

Benchscale Assessment of the Efficacy of a Reactive Core Mat to Isolate PAH-Spiked Aquatic Sediments

This paper describes the results of a benchscale testing program to assess the efficacy of a reactive core mat (RCM) for short-term isolation and partial remediation of contaminated, subaqueous sediments. The 1.25-cm-thick RCM (with a core reactive material such as organoclay with filtering layers on top and bottom) is placed on the sediment, and approximately 7.5–10 cm of overlying soil is placed on the RCM for stability and protection. A set of experiments were conducted to measure the sorption characteristics of the mat core (organoclay) and sediment used in the experiments, and to determine the fate of semi-volatile organic contaminants and non-reactive tracers through the sediment and reactive mat. The experimental study was conducted on naphthalene-spiked Neponset River (Milton, MA) sediment. The results show nonlinear sorption behavior for organoclay, with sorption capacity increasing with increasing naphthalene concentration. Neponset River sediment showed a notably high sorption capacity, likely due to the relatively high organic carbon fraction (14%). The fate and transport experiments demonstrated the short-term efficiency of the reactive mat to capture the contamination that is associated with the post-capping period during which the highest consolidation-induced advective flux occurs, driving solid particles, pore fluid, and soluble contaminants toward the reactive mat. The goal of the mat placement is to provide a physical filtering and chemically reactive layer to isolate contamination from the overlying water column. An important finding is that, because of the high sorption capacity of the Neponset River sediment, the physical filtering capability of the mat is as critical as its chemical reactive capacity.     

Efficient direct chromosome analyses and enzyme determinations from chorionic villi samples in the first trimester of pregnancy

Chorionic villi were obtained by an aspiration technique which proved to be the best of four alternative procedures. We report in detail the series of experiments which led to (1) successful, rapidly growing cell cultures practically free of maternal cell contamination (the use of hormone-supplemented Chang medium greatly increased the growth rate); (2) an efficient direct method to obtain high quality metaphases from the Langhans cells of the cytotrophoblast tissue and with which the fetal karyotype is defined within a few hours of chorionic villi sampling; and (3) successful testing for the activity of eight enzymes directly from the villi samples, thus showing that this material is suitable for a rapid, direct diagnosis of the related metabolic diseases.