短片段基因序列测定检测沙门菌

[目的]通过对沙门菌fimy基因保守片段的焦磷酸测序,达到鉴定沙门菌的目的。[方法]通过短片段基因序列测定检测沙门菌。[结果]通过焦磷酸测序法对fimy基因上的27个碱基片段进行测定,可准确检测191株沙门菌。[结论]建立的方法具有准确、快速和实时检测等优点,适于对沙门菌的快速、高通量检测。     

应用PCR技术快速检测沙门菌

[目的]建立用于快速和特异检测沙门菌的PCR方法。[方法]将受试菌种按煮沸裂解法提取DNA,针对hns与invA基因序列合成2对PCR引物,对沙门菌标准菌株和其他菌属的菌株进行PCR检测,并用福建省2006—2007年沙门菌临床分离株137株进行验证。[结果]沙门菌标准菌株hns和invA基因均为阳性;137株沙门菌临床分离株hns和invA基因的阳性率分别为100％和99．3％,其他菌属的菌株无特异性条带。[结论]本研究建立的PCR方法可用于检测沙门菌属。     

复合PCR鉴定沙门菌的方法

沙门菌属于肠杆菌科，是具有鞭毛、能运动的革兰阴性杆菌，而且它是导致食源性疾病的主要病原菌之一。为了加强口岸执法管理、缩短检验检疫流程和提高检验检疫工作效率，本实验拟建立复合聚合酶链反应快速检验鼠伤寒沙门菌的方法，以求提高检验工作准确性和速度。本实验在参考国内外文献的基础上，设计了两对引物，建立了较稳定的PCR检测系统。本实验中，使用了不同的菌株作为比较，证实了系统特异性良好，并且从不同的食品来源检出的沙门菌，皆能被本实验系统鉴定检出。使用复合PCR技术，能快速、准确的鉴定肠炎沙门菌和鼠伤寒沙门菌，并且较之经典的鉴定方法，时间上提前了2—3d。     

基于ＴａｑＭａｎ探针三重荧光ＰＣＲ 检测沙门菌、肠炎沙门菌和鼠伤寒沙门菌

摘要：目的　建立基于ＴａｑＭａｎ探针三重荧光ＰＣＲ 检测沙门菌（狊犪犾犿狅狀犲犾犾犪,Ｓａ）、肠炎沙门菌（ＳＥ）和鼠伤寒沙门菌（ＳＴ） 的方法。方法　根据沙门菌犪犮犲犃基因、肠炎沙门菌特异序列（ＧｅｎＢａｎｋ：ＡＦ３７０７０７．１）、鼠伤寒沙门菌的ＳＴＭ４５９９ 序列（ＧｅｎＢａｎｋ：ＡＥＲＶ０１００００２３．１）,分别设计引物和ＴａｑＭａｎ探针,在探针的５′端分别标记ＦＡＭ、ＶＩＣ、ｃｙ５,建立基于ＴａｑＭａｎ探针三重实时荧光ＰＣＲ 检测沙门菌的方法。结果　２９ 种不同血清型沙门菌均扩增出犪犮犲犃基因,肠炎沙门菌和鼠伤寒沙门菌的引物和探针分别特异性地扩增出１５株肠炎沙门菌和１１株鼠伤寒沙门菌,而其他血清型沙门菌和１７株非沙门菌扩增结果阴性。犪犮犲犃、肠炎沙门菌、鼠伤寒沙门菌的三重荧光ＰＣＲ 扩增效率分别为８９％、８７％、９０％,最低检测浓度分别达到２８０ｃｆｕ／ｍｌ、２６０ｃｆｕ／ｍｌ、３００ｃｆｕ／ｍｌ。结论　本研究建立的方法特异性好、灵敏度高,可用于食品中沙门菌、肠炎沙门菌和鼠伤寒沙门菌的特异性检测。     

环介导等温扩增（LAMP）技术检测沙门菌属方法的建立

建立一种检测沙门菌属快速敏感的环介导等温扩增（loop-mediated isothermal amplification，LAMP）检测方法。针对沙门菌属侵袭蛋白（invA）基因序列的六个区域设计四条LAMP引物，65℃保温约60min，完成对沙门菌属的扩增，扩增产物经电泳和酶切鉴定。利用LAMP和普通PCR方法同时检测4株沙门菌和9株非沙门菌（对照组）来验证LAMP方法的特异性；将肠炎沙门菌菌液做一系列10倍稀释后用LAMP和PCR方法进行检测来比较两者敏感性。4株沙门菌扩增出LAMP特征性梯状条带，9株非沙门菌没有出现LAMP扩增，LAMP产物的特异性通过限制性内切酶得到了证实；LAMP检测沙门菌属的特异性与普通PCR相同，但其敏感性比普通PCR高10倍，LAMP检测沙门菌属的检测下限为10cfu／ml，PCR检测下限为100cfu／ml。LAMP检测速度相比PCR更快速，在60min内即可完成扩增反应。建立了一个快速、特异、敏感的沙门菌属LAMP检测方法。     

基于TaqMan探针双重荧光PCR检测食品中鼠伤寒沙门菌和肠炎沙门菌

目的建立基于TaqMan探针双重荧光PCR检测鼠伤寒沙门菌和(Salmonella typhimurium,ST)和肠炎沙门菌(Salmonella enteritidis,SE)的方法。方法根据ST的STM4599序列(GenBank:AERV01000023.1)和SE特异序列(GenBank:AF370707.1),分别设计引物和探针,ST探针的5'端标记FAM、SE探针的5'端标记VIC,建立基于TaqMan探针双重荧光PCR检测方法。结果 ST和SE的引物和探针分别特异性地扩增出16株ST和15株SE,而28种不同血清型沙门菌和17株变形杆菌等扩增结果均为阴性。ST和SE的双重荧光PCR扩增效率均为94.2%,R2分别为0.998和0.995,最低检测浓度分别达到300 CFU/ml、260 CFU/ml。结论建立的方法特异性好、灵敏度高,整个试验可在31h完成,是快速检测ST和SE的有效方法,可用于食品中ST和SE的特异性检测。     

PCR检测沙门菌hilA基因的方法研究

目的:利用PCR技术,建立特异性引物PCR快速检测沙门菌的方法。方法:设计沙门菌h ilA基因的特异性引物,优化PCR反应条件,对沙门菌和非沙门菌进行特异性引物的PCR扩增,鉴定沙门菌。结果:所检测沙门菌株和模拟样品均出现特异性扩增条带,结果与实际相符。结论:该方法具有简单、迅速、特异性好和敏感性高的特点。     

一起由肠炎沙门菌和奇异变形杆菌引起的食物中毒

[目的]食物中毒病原菌的检测鉴定。[方法]采集食物中毒肛拭、可疑食物、涂抹标本共18份,按照GB/T4789.4-2003和WS/T9-1996进行检测。[结果]其中7份肛拭中同时检出肠炎沙门菌和奇异变形杆菌,在菜勺、菜盆、操作台4份涂抹标本中检出奇异变形杆菌。[结论]此次食物中毒是由肠炎沙门菌和奇异变形杆菌混合感染所致。     

多重PCR法鉴定伤寒沙门菌的研究

目的建立多重PCR方法鉴定伤寒沙门菌,为临床确诊及现场流行病学调查提供快速准确的实验室技术支持。方法根据沙门菌菌体抗原A/D1群基因、鞭毛抗原基因(fliC-Hd)及伤寒沙门菌Vi抗原基因片段(Vi)设计引物,建立多重PCR体系并进行反应条件优化。选择15株不同血清型沙门菌及18株非沙门菌菌株,对所建立体系的特异性进行检测,并将该体系应用于浙江省分离的50株实际样本的检测。结果建立并优化了伤寒沙门菌检测的多重PCR体系,优化后25μl PCR体系包括100μM dNTPs、2.5 U Taq DNA聚合酶、引物各0.2μM、模板5μl;PCR条件为94℃变性1 min,56℃退火1 min,72℃延伸1 min,共循环35次。该体系具有高特异性,可准确快速鉴定伤寒血清型沙门菌,同时也可区分A群及D群血清群的沙门菌,并能检测鞭毛抗原为Hd及毒力基因Vi阳性的沙门菌,对实际样本检测符合率达100.00%。结论多重PCR可准确鉴定伤寒沙门菌,能作为传统血清学分型的辅助方法用于伤寒沙门菌的鉴定。     

脉冲场凝胶电泳在沙门菌食物中毒调查检验中的应用

[目的]为肠炎沙门菌（Salmonella enteritidis）食物中毒的判定提供实验室检测依据。[方法]采用脉冲场凝胶电泳（PFGE）对1起食物中毒病人分离株和福建省以往分离的肠炎沙门菌株进行分子分型。[结果]不同病人的6株肠炎沙门菌PFGE结果均为同一型别,与往年分离的病人分离株的型别有异。[结论]PFGE分型技术可用于肠炎沙门菌的准确分型,有助于食物中毒的流行病学分析、溯源、判定和控制。     

Calcium intake in the elderly: validation of a dietary questionnaire

Epidemiological studies of calcium and osteoporosis have been hampered by the lack of a suitable tool for assessing calcium intake. This report describes a new frequency and amount questionnaire for measuring present and past calcium intake in the elderly. The validity of the questionnaire was tested against two commonly used standards of dietary assessment, five-day duplicate diets and seven-day weighed dietary inventories. The resulting correlation coefficients were, respectively, r = 0.76 and r = 0.69, while that for repeatability was r = 0.84. Furthermore, the questionnaire categorized subjects into thirds of the distribution of intake with almost no gross misclassification. It is suggested that the present findings may be extended to the majority of normal, healthy elderly subjects, implying wide application for the questionnaire in the assessment of calcium intake in the elderly.     

The occurrence of antimicrobial substances in toilet,sink and shower drainpipes of clinical units: A neglected source of antibiotic residues

Antibiotics represent one of the most important drug groups used in the management of bacterial infections in humans and animals. Due to the increasing problem of antibiotic resistance, assurance of the antibacterial effectiveness of these substances has moved into the focus of public health. The reduction in antibiotic residues in wastewater and the environment may play a decisive role in the development of increasing rates of antibiotic resistance. The present study examines the wastewater of 31 patient rooms of various German clinics for possible residues of antibiotics, as well as the wastewater of five private households as a reference.To the best of our knowledge, this study shows for the first time that in hospitals with high antibiotic consumption rates, residues of these drugs can be regularly detected in toilets, sink siphons and shower drains at concentrations ranging from 0.02?μg·L^?1 to a maximum of 79?mg·L^?1. After complete flushing of the wastewater siphons, antibiotics are no longer detectable, but after temporal stagnation, the concentration of the active substances in the water phases of respective siphons increases again, suggesting that antibiotics persist through the washing process in biofilms. This study demonstrates that clinical wastewater systems offer further possibilities for the optimization of antibiotic resistance surveillance.     

Self-reported health problems among Swedish miners one year after unemployment

Unemployment is considered to be a public health concern sincedeterioration in the health of the unemployed is often anticipated.However, for some groups, such as miners, unemployment mightimprove health due to a cessation of potentially harmful occupationalexposures. This study evaluates the health of 79 miners in oneSwedish iron-ore mine, and 226 age-matched controls from thegeneral population, during one year after the closure of themine. The participants received a questionnaire regarding medicalhistory and subjective symptoms at the beginning of the studyperiod, and after one year. Statistically significant negativeeffects on self-reported health attributable to unemploymentwere not found, although neuropsychiatric symptoms were morecommon among the unemployed miners. The miners reported a statisticallysignificant improvement in grip force (p=0.031). They had asignificantly higher prevalence of symptoms associated withmining related exposures when compared with the population controls;pain in the upper extremities [relative risk (RR)=2.27, 95%confidence interval (Cl)=1.44–3.59), back pain (RR=1.84;Cl=1.237–2.75), vasospastic disease of the fingers (RR=2.05;Cl=1.18–3.57) and obstructive respiratory symptoms (attacksof dyspnea and wheezing: RR=3.67; Cl=1.167–11.6).     

Excès de cas humains et animaux de tularémie en France au cours de l’hiver 2007–2008 : émergence ou phénomène isolé ?

Context

Tularemia is a zoonosis affecting humans and hares in France. We describe the results of surveillance in both species, in 2007 and 2008.

Methods

Human tularemia cases are mandatorily notifiable in France since 2003. In hares, surveillance relies on volunteer hunter associations in all districts of the country. Data from mandatory reports and volunteer surveillance in 2007/2008 were analyzed and compared with previous results.

Results

In 2007/2008, 144 cases were reported in humans and 117 cases in hares. This was a 100% increase compared to previous years. Human cases differed from those of previous years only by the frequency of contact with breeding animals. Human cases without any documented risk exposure were also more frequent.

Conclusion

An increase of tularemia cases occurred in 2007/2008 in both species. Complementary studies are needed to identify the species reservoir in France to understand the causes of this peak of cases.     

Occupational health hazards in mining: an overview

This review article outlines the physical, chemical, biological, ergonomic and psychosocial occupational health hazards of mining and associated metallurgical processes. Mining remains an important industrial sector in many parts of the world and although substantial progress has been made in the control of occupational health hazards, there remains room for further risk reduction. This applies particularly to traumatic injury hazards, ergonomic hazards and noise. Vigilance is also required to ensure exposures to coal dust and crystalline silica remain effectively controlled.     

Manufacture and use of nanomaterials: current status in the UK and global trends

This paper provides an overview of the production and use of nanomaterials (NMs), particularly in the UK. Currently, relatively few companies in the UK are identifiable as NM manufacturers, the main emphasis being the bulk markets in metals and metal oxides, and some niche markets such as carbon nanotubes and quantum dots. NM manufacturing in the UK does not reflect the global emphasis on fullerenes, nanotubes and fibres. Some assumptions have been made about the types of NM that are likely to be imported into the UK, which currently include fullerenes, modified fullerenes and other carbon-based NMs including nanotubes. Many university departments, spin-offs and private companies have developed processes for the manufacture of NMs but may only be producing small quantities for research and development (R&D) purposes. However, some have the potential to scale up to produce large quantities. The nanotechnology industry in the UK has strong R&D backup from universities and related institutions. This review has covered R&D trends at such institutions, and appropriate information has been added to a searchable database. While several companies are including NMs in their products, only a few (e.g. manufacturers of paints, coatings, cosmetics, catalysts, polymer composites) are using nanoparticles (NPs) in any significant quantities. However, this situation is likely to change rapidly. There is a need to collect more information about exposure to NPs in both manufacturing and user scenarios. As the market grows, and as manufacturers switch from the micro- to the nanoscale, the potential for exposure will increase. More research is required to quantify any risks to workers and consumers.     

深圳公立医院管理体制改革制度设计

深圳公立医院管理体制改革实行政事分开、管办分开,在理事会架构下按法定机构模式组建市医管中心,落实公立医院运营管理自主权.作者从当前公立医院管理体制的弊端入手,介绍了深圳市进行公立医院管理体制改革的基本思路及改革方案设计的主要举措,深入剖析了的改革方案的特点,并对改革效果进行了预测.     

Effects of nitrogen dioxide on red blood cells of rats: Changes in components of red cell membranes during in vivo exposure to NO2

Red cell membranes, prepared from red blood cells of rats exposed to 4, 10, or 20 ppm nitrogen dioxide (NO₂) for 1 to 10 days, were examined for evidence of changes in membrane components. Appreciable changes were not found in contents of phospholipid and cholesterol during exposure to 10 ppm NO₂. By contrast, protein content altered with the time of exposure. Moreover, changes in protein composition were observed by employing sodium dodecyl sulfate — polyacrylamide gel electrophoresis. Twenty-four-hour exposure to NO₂ at the concentration above 10 ppm resulted in a marked increase in the percentage of lysophosphatidylethanolamine (LysoPE) to the total phospholipids. The prolonged exposure to 10 ppm NO₂ gave rise to a further increase in LysoPE, whereas the percentage of phosphatidylethanolamine (PE) showed a gradual decrease. A 1-day exposure to 4.0 ppm NO₂ also caused an increase in sialic acid content and decreases in those of PE and hexose. In addition to contents of these components the percentage of LysoPE increased 5 days after exposure and the elevated values were maintained up to the end of exposure period. These results demonstrate that red blood cells in circulation exhibit different membrane properties in terms of lipid and carbohydrate composition during 10 days of exposure to 4.0 ppm NO₂.     

Investigating clusters in the workplace and beyond

Clusters of disease are common and occur in the workplace and in the general community. They often arouse considerable concern among the population. Investigations have sometimes lead to exciting new knowledge, but in general the investigation of clusters is difficult and often unrewarding, especially for community clusters. In the workplace, investigations are more likely to find associations and even new causes, but still many clusters remain enigmatic. Despite this, there are many reasons for investigating clusters, including allaying community concern and identifying uncontrolled exposures. A structure for investigating clusters in the workplace is suggested.