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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

通用型基因悬浮芯片检测生物恐怖细菌的方法研究

Objective To develop a fast, high-throughput screening method with suspension array technique for simultaneous detection of biothreat bacteria. Methods 16 S rDNA universal primers for Bacillus anthracis, Francisella tularensis, Yersinia pestis, Brucella spp. and Burkholderia pseudomallei were selected to amplify corresponding regions and the genus-specific or species-specific probes were designed. After amplification of chromosomal DNA by 16 S rDNA primers 341A and 519B,the PCR products were detected by suspension array technique. The sensitivity, specificity, reproducibility and detection power were also analyzed. Results After PCR amplification by 16 S rDNA primers and specific probe hybridization,the target microorganisms could be identified at genus level, cross reaction was recognized in the same genus. The detection sensitivity of the assay was 1.5 pg/μl (Burkholderia pseudomallei),20 pg/μl (Brucella spp.), 7 pg/μl (Bacillus anthracis), 0.1 pg/μ1 (Francisella tularensis), and 1.1 pg/μl (Yersinia pestis),respectively. The coefficient of variation for 15 test of different probes was ranged from 5.18% to 17.88%,it showed good reproducibility. The assay could correctly identify Bacillus anthracis and Yersinia pestis strains in simulated white powder samples. Conclusion The suspension array technique could be served as an opening screening method for biothreat bacteria rapid detection.     

对医院护理管理队伍建设的几点思考

文章分析了当前医院护理管理队伍存在的主要问题：知识结构不合理；专业思想不纯正；接受继续教育不足：工作效能低下。提出了建设高素质护理管理人才队伍的思路：严格标准，搞好选才；加强思想教育和引导；加大培养力度；建立科学考评和激励机制；完善人才合理流动措施；合理编制，突出效率。     

政府公共财政支出分配公平性研究进展

政府公共财政支出是社会再分配的一种形式，其目的是促进社会公平。因此，政府的公共财政支出应该具有较强的目标针对性，应该向社会贫困人群倾斜，使其从中受益。本文通过查阅国内外政府公共财政支出分配公平性的相关文献资料。阐述了相关研究的进展情况，井着重探讨了政府公共财政支出分配公平性的内涵、研究范围、测算方法和研究意义。以及实际研究中存在的问题，最后提出我国政府公共财政支出分配公平性的研究方向和需要进一步完善的地方。     

长春新碱过量引起严重毒副反应1例的护理体会

报道1例霍奇金淋巴瘤患者因误用长春新碱（VCR）10mg一次性静脉推注后治疗护理情况。其出现间断性神志恍惚、眼睑闭合不全、言语不清、口腔黏膜糜烂、全身疼痛、麻痹性肠梗阻、尿潴留、手足麻木等症状,经积极解救,禁食,持续胃肠减压、胃管内注入麻油、开塞露、生理盐水灌肠,合理应用肠外营养,注重疼痛、心理护理,做好口腔、肛周护理,预防感染加重,患者病情得到控制好转出院。     

Problem of methodology of evaluation of the quality of public health

The basic general methodological assessments of public health quality are considered. An analogy is drawn with quality assessment in industry, approaches to the development of multi-aspect and multi-parameter assessment of public health quality are substantiated. A term of "qualimetry " of public health is suggested.     

上海市某医院骨科平均住院日影响因素分析

对上海市某医院2003年-2007年骨科出院病人的住院日描述性分析．2003年-2007年骨科的床位利用指数与平均住院日相关性分析．2003年-2007年骨科床位与医护比例分析．2007年骨科前10大病种平均住院目影响因素分别进行单因素相关性分析和多因素逐步回归分析（STATA软件）。通过对骨科10大病种住院日影响因素分析,术前等待天数、手术类型、是否输血分别对10个、9个和8个病种的住院目有影响。输血因素和手术类型是医院不可控、由病人的病情决定的,术前等待天数是管理因素,是最值得医院重视的影响因素。