嗜水气单胞菌TaqMan实时PCR检测方法的建立

Objective To develop a TaqMan real-time PCR for the detection of aeromonas hydrophila. Methods The conserved region of major adhesion gene of aeromonas hydrophila (aha) was used to design primers and TaqMan probe. A total of six concentration gradients for forward and reverse primers ranging from 200 -700 nmol/L were chosen, and four concentration gradients for probe ranging from I00 -400 nmol/L were chosen. And then the concentration of primers and probe were optimized by ANOVA of completely randomized design respectively. The specificity of the established method was evaluated by using bacteria as contrast, including 45 strains Vibrio cholerae,20 strains Vibrio parahemolyticus, 10 strains Vibrio fluvialis,4 strains Vibrio mimicus,5 strains Vibrio vulnificus, 1 strain Vibrio aiginoayticns, 1 strain Vibrio furnissii, 5 strains Salmonella, 10 strains Shigella and 2 strains Piesiomonas shigelloides. The sensitivity, bacterial sensitivity and DNA sensitivity included,were evaluated. The stool of healthy people was contaminated by aeromonas hydrephila artificially, and the ability of the established TaqMan real-time PCR system for detection of aeromonas hydrophila was also evaluated. Results The cycle threshold (Ct) value deserved from 6 groups of primers concentration gradient was (x±s) :20.69±0.33,20.72±0.21,20.81±0. 12,20.74±0.12,20.51±0. 16 and 20.69±0. 11, respectively, and the concentration of forward primer and reverse primer was determined to be 200 nmol/L (F=1.33, P=0. 28). The Ct value deserved from 4 groups of probe concentration gradient was (x±s) : 20.56±0. 08,20.82±0.05,20. 82±0. 11 and 20.93±0.09,respectively,and the concentration of probe was determined to be 100 nmol/L(F =5.26,P =O. 01 ). The bacterial sensitivity and DNA sensitivity were 80 CFU/ml and 100 fg/μl respectively,and the sensitivity to detect aeromonas hydrophila from stool was 8 × 103 CFU/ml, which might be 8 CFU/ml after 8 hours' enrichment. No amplification was observed in the templates of other bacterial. Conclusion The TaqMan real-time PCR method targeting the aha gene of aeromonas hydrophila had a high sensitivity and specificity and might be used to detect aeromonas hydrophila from pure bacterial and stool rapidly.     

山东省4类人群对艾滋病的知识、态度抽样调查

[目的]了解山东省4类人群对艾滋病的知识认识现状，明确现阶段的重点宣传对象与宣传内容。[方法]2002年10月至2003年3月，在山东省全部17个市，采用多阶段整群抽样和配额抽样相结合的方法，对4类人群共3234名进行问卷调查。[结果]4类人群对艾滋病的知识都有一定的了解，机关人员、特殊人群、城镇居民和大中学生的总体知晓率分别为84．9％、79．1％、81．6％和78．8％，但对一些问题的认识存有偏差。其中机关人员和文化程度较高者知晓率较高。特殊人群、大中学生以及文化程度较低的人群知道得较少。[结论]4类人群对艾滋病的知识掌握不够系统、全面，应加强对特殊人群、大中学生以及文化程度较低人群的健康教育工作。     

赴南非和泰国考察艾滋病的报告

南非、泰国在艾滋病预防控制和管理方面的经验教训很值得我们借鉴。2003年11月11～21日，笔者有幸随山东省卫生厅考察团赴这两国考察，颇有收获。现将两国艾滋病防治工作的开展情况介绍如下。     

山东省6县（区）部分人群预防SARS行为调查分析

目的了解山东省不同地域居民SARS流行期生活行为及流行前后行为变化情况。方法选择城市和乡镇社区对于SARS传播和控制意义较重要的具有代表性的7个人群进行入户式现场调查。结果人们从个人卫生、公共卫生、提高身体素质、减少接触机会、维护生态平衡等多个方面约束或改进了个人行为。92.0%的被调查者认为非典流行促使自己改进了生活习惯。结论山东省人群非典期间采取预防行为的比率很高,为今后SARS、禽流感及其他呼吸道传染病的预防奠定了良好的基础。     

山东省艾滋病高危人群规模估计及疫情预测

[目的]估计山东省艾滋病各类高危人群的规模，预测艾滋病病毒（HIV）感染者人数，为艾滋病防治工作提供依据。[方法]依据山东省艾滋病病例报告、哨点监测、艾滋病专题调查等获得的数据，参考有关资料，运用Workbook估计法，估计山东省各市艾滋病高危人群基数，预测艾滋病疫情。[结果]估计山东省现有HIV高危险人群1023770～1864181人；预测山东省现有艾滋病感染者7077人。[结论]山东省目前仍处于艾滋病低流行状态，但流行形势并不乐观。     

山东省艾滋病感染者子女现况调查

[目的]了解山东省艾滋病感染者子女的分布和生存状况。[方法]2005年3～4月，对山东省全部HIV感染者的18岁及其以下子女的一般情况、就学情况、HIV感染状况及其家庭的相关情况进行了调查：[结果]调查HIV感染者子女208人，分布在13个市、38个县（市、区）、142个家庭；做过HIV抗体筛查的128人，阳性1人；46人因家庭经济困难而辍学，调查142个家庭，28个家庭有2名HIV感染者，1个有3名HIV感染者;95个家庭年人均收入低于1000元。[结论]HIV感染者子女分布比较广泛，受教育情况、家庭经济状况相对较差，政府和社会有关部门应为他们提供更多的关怀和帮助。     

A群链球菌病原学与流行病学研究进展

A群链球菌（GAS）所致疾病谱广泛，轻者如咽炎、扁桃体炎、猩红热等，重者如坏死性筋膜炎、急性肾小球肾炎、链球菌中毒性休克综合征（STSS）等。本文从GAS的病原学和流行病学两方面进行综述。     

嗜水气单胞菌TaqMan实时PCR检测方法的建立

Objective To develop a TaqMan real-time PCR for the detection of aeromonas hydrophila. Methods The conserved region of major adhesion gene of aeromonas hydrophila (aha) was used to design primers and TaqMan probe. A total of six concentration gradients for forward and reverse primers ranging from 200 -700 nmol/L were chosen, and four concentration gradients for probe ranging from I00 -400 nmol/L were chosen. And then the concentration of primers and probe were optimized by ANOVA of completely randomized design respectively. The specificity of the established method was evaluated by using bacteria as contrast, including 45 strains Vibrio cholerae,20 strains Vibrio parahemolyticus, 10 strains Vibrio fluvialis,4 strains Vibrio mimicus,5 strains Vibrio vulnificus, 1 strain Vibrio aiginoayticns, 1 strain Vibrio furnissii, 5 strains Salmonella, 10 strains Shigella and 2 strains Piesiomonas shigelloides. The sensitivity, bacterial sensitivity and DNA sensitivity included,were evaluated. The stool of healthy people was contaminated by aeromonas hydrephila artificially, and the ability of the established TaqMan real-time PCR system for detection of aeromonas hydrophila was also evaluated. Results The cycle threshold (Ct) value deserved from 6 groups of primers concentration gradient was (x±s) :20.69±0.33,20.72±0.21,20.81±0. 12,20.74±0.12,20.51±0. 16 and 20.69±0. 11, respectively, and the concentration of forward primer and reverse primer was determined to be 200 nmol/L (F=1.33, P=0. 28). The Ct value deserved from 4 groups of probe concentration gradient was (x±s) : 20.56±0. 08,20.82±0.05,20. 82±0. 11 and 20.93±0.09,respectively,and the concentration of probe was determined to be 100 nmol/L(F =5.26,P =O. 01 ). The bacterial sensitivity and DNA sensitivity were 80 CFU/ml and 100 fg/μl respectively,and the sensitivity to detect aeromonas hydrophila from stool was 8 × 103 CFU/ml, which might be 8 CFU/ml after 8 hours' enrichment. No amplification was observed in the templates of other bacterial. Conclusion The TaqMan real-time PCR method targeting the aha gene of aeromonas hydrophila had a high sensitivity and specificity and might be used to detect aeromonas hydrophila from pure bacterial and stool rapidly.     

肾综合征出血热病毒基因型和流行特点分析

目的探讨山东省汉坦病毒(HV)主要流行株的基因型和流行特点。方法采集山东地区有代表性的鼠肺标本，应用逆转录聚合酶链反应(RT—PCR)扩增技术及核苷酸序列测定方法对免疫荧光抗原阳性的标本进行HV G2基因扩增并测序，与原有山东毒株一起进行同源性分析、系统发生树分析。结果山东省新分离18株HV均为汉城(SEO)型，与以往山东SEO型HV有较高同源性，在95．7％以上，属于同一亚型。山东省汉滩(HTN)型HV同源性较低，在77．9％以上，属于不同亚型。结论山东省HV是以SEO型为主的SEO和HTN混合型疫区。SEO型基因变异率低，有较高的稳定性。而HTN型HV变异率高，稳定性差。     

肠聚集性大肠杆菌毒力因子的研究进展

肠聚集性大肠杆菌(EAggEC)是近年确认的一种致腹泻大肠杆菌,与儿童腹泻密切相关.在印度、墨西哥及巴西等地进行的前瞻性研究表明,EAggEC是病程超过2周的持续性腹泻的重要病原之一,并能导致儿童营养不良.最近还有报道EAggEC与旅游者腹泻有关.发现EAggEC还是HIV感染者慢性腹泻的病原菌之一.但是,关于其致病机理尚不十分清楚.粘附性和粘附因子被认为是其致病的重要因素.还有证据表明EAggEC能产生耐热和不耐热毒素.对细胞具有侵袭性、能引起接触性溶血.本文对近年内EAggEC致病因子的研究现状综述如下.1 粘附性和粘附因子致泻的大肠杆菌对组织培养的上皮细胞株如HEp—2、Hela等表现为3种形式的粘附,即局灶性粘附(LA)、弥散性粘附(DA)和聚集性粘附(AA).LA