胶体金免疫层析法检测水产品中O1群霍乱弧菌方法的建立与优化

〔目的〕研究了O1群霍乱弧菌免疫层析试纸的制备方法,通过对试纸材料的处理达到优化胶体金免疫层析体系的目的。〔方法〕建立了快速检测食品中O1群霍乱弧菌的胶体金免疫层析方法,并应用该方法对弧菌属、假单胞菌属及其他常见肠道致病菌,共29株进行检测,同时对180份水产品进行了样品添加试验,并用《出口食品中霍乱弧菌检验方法》(SN/T1022-2001)进行确证,全面评估了该方法的灵敏度、特异性、准确性等。〔结果〕该方法检测食品中O1群霍乱弧菌的灵敏度为105cfu/ml,与食品中常见的致病菌无交叉反应。该方法检测水产品中O1群霍乱弧菌的假阳性率为6.25%。〔结论〕该方法操作简便,灵敏度高,特异性强,准确性高,适用于食品的快速检测。     

利用胶体金免疫层析技术快速定量检测单增李斯特菌方法的建立

〔目的〕建立胶体金免疫层析技术快速定量检测单增李斯特菌方法。〔方法〕利用胶体金标记和双抗体夹心免疫层析技术,建立单增李斯特菌的快速检测方法,评价其特异性和敏感性,并拟合标准曲线进行定量检测;在牛奶、橙汁、果冻等即食食品样品中添加单增李斯特菌模拟污染样品,评价该方法对半固体、液体等即食食品、可疑生物恐怖样品的检测能力。〔结果〕该法可在10min内完成定性和半定量检测,定量检测灵敏度为3.5×103cfu/ml,线性范围3.5×105～3.5×108cfu/ml,回收率在99%～101.7%之间。〔结论〕建立的检测单增李斯特菌的胶体金免疫层析方法,能快速、灵敏、特异、准确地对样品中的单增李斯特菌进行定性、定量检测。     

建立快速检测鼠疫耶尔森菌的胶体金免疫层析法

〔目的〕　建立检测鼠疫耶尔森菌 (鼠疫杆菌 )的胶体金标记免疫层析方法。〔方法〕　采用柠檬酸三钠还原法制备胶体金颗粒 ,并标记鼠疫杆菌的抗体 ,制成免疫层析检测试纸条 ,进行特异性和敏感性评价 ,并对面粉、土壤、奶粉、淀粉等不同材料进行模拟添加鼠疫菌检测。〔结果〕　用多种不同的常见细菌或近缘细菌进行测试 ,未出现非特异性反应 ,检测鼠疫杆菌的灵敏度为 1× 10 5cfu/ml,对水、土壤、奶粉、面粉、淀粉、酵母粉等不同材料中鼠疫杆菌进行检测也取得同样结果 ,10～ 15min即可得到结果。〔结论〕　建立了 1种可以用于现场快速检测鼠疫杆菌的胶体金免疫层析方法 ,为进一步大规模应用奠定了基础。     

斑点免疫金渗滤法检测沙门菌的研究与应用

〔目的〕研究一种特异性强,灵敏度高,且操作简便,快速的沙门菌检测方法,以用于临床样品中沙门菌的快速检测,并能在基层推广应用。〔方法〕应用胶体金标记纯化的兔抗沙门菌抗体-1,SPA(金黄色葡萄球菌A蛋白)包被硝酸纤维素膜作对照点,建立直接检测沙门菌的斑点免疫金渗滤法(DIGFA)检测试纸盒。〔结果〕通过胶体金标记兔抗沙门菌抗体直接显色,阳性者出现红色斑点,结果易于判断。整个试验过程仅需10￣15min即可判断结果,操作简单,与大肠埃希菌、枸橼酸杆菌、变形杆菌和绿脓杆菌等不发生交叉反应。沙门菌的最低检测量为4.5×106cfu/ml。同时将该法与国标法检测效果比较,符合率达100%。将胶体金标记的兔抗沙门菌抗体冻干粉4℃存放12个月,检测结果无差异。〔结论〕研究结果表明该法具有微量、特异、敏感、快速可靠,效果直观,判断容易,无需特殊仪器设备,非常适用于沙门菌的快速检测,值得推广应用。     

应用胶体金免疫层析技术建立炭疽杆菌芽孢的快速检测方法

〔目的〕　建立 1种炭疽杆菌芽孢的快速检测方法。〔方法〕　利用胶体金免疫层析技术 ,采用双抗体夹心法检测炭疽芽孢杆菌 ,对该方法进行特异性、敏感性等评价 ;在面粉、淀粉、奶粉等材料中掺入炭疽疫苗株芽孢模拟“白色粉末” ,评价该法检测炭疽生物恐怖和环境样品的可行性。〔结果〕　该法能在 2 0min内完成检测 ,多种不同的芽孢菌及其它菌检测评价显示该法特异性良好 ,检测灵敏性为 1× 10 6cfu/ml,环境样品的检测敏感性也相同。〔结论〕　建立的检测炭疽芽孢杆菌的胶体金免疫层析方法 ,能快速、特异、灵敏的检测炭疽芽孢 ,适用于现场的快速检测。     

大肠杆菌O157胶体金免疫层析快速筛查方法的建立

目的建立一种大肠杆菌O157的胶体金免疫层析快速筛查方法。方法利用胶体金免疫层析技术,采用双抗体夹心法检测大肠杆菌O157,对该法进行敏感性、特异性和食品样品的适用性分析。结果该法能在15分钟内完成检测,检测不同的肠杆菌科细菌(非O157型大肠杆菌、沙门菌、志贺菌、变形杆菌、柠檬酸杆菌、肠杆菌、沙雷菌、耶尔森菌)、葡萄球菌、李斯特菌、气单胞菌、弧菌等共24种30株常见细菌,未发现有交叉反应,显示该法特异性良好。检测大肠杆菌O157的最低检出浓度为1×105cfu/ml。方法适用于奶粉、面粉、淀粉、咖啡粉、饼干、蛋糕、果冻、燕窝和果汁等食品样品的检测。结论新建立的大肠杆菌O157胶体金免疫层析试验简便、快速,特异性和灵敏度较好,适用于现场样品的快速筛查。     

快速检测O9抗原沙门菌免疫金标层析法的建立及其应用研究

目的:建立一种快速检测O9抗原沙门菌的新方法。方法:利用针对沙门菌O9抗原的两株特异性单克隆抗体,建立一种双抗体夹心检测O9抗原沙门菌的胶体金免疫层析试纸条(GICA)。结果:该试纸条对标准肠炎沙门菌LPS的最小检出量为0.15 ng/条,对肠炎沙门菌菌液的最小检出量为2×105cfu/条。蛋品、鸡肉、猪肉的人工污染试验显示,经增菌后免疫层析试纸条可检出1 cfu的肠炎沙门菌,在大肠杆菌与沙门菌比例为1000:1仍不干扰试纸条的检测。共对市场采集的鸡蛋、鸡肉以及猪肉样品577份进行免疫层析试纸的检测,试纸条与经典细菌分离鉴定法相比:敏感度为90.0%,特异性为98.6%。结论:检测O9抗原沙门菌的GICA法快速、特异、灵敏,在重要沙门菌病检测方面有着潜在、广阔的应用前景。     

一种快速筛查金黄色葡萄球菌的荧光纳米免疫层析方法的建立

目的建立一种快速筛查金黄色葡萄球菌的荧光纳米免疫层析检测方法。方法以荧光纳米颗粒作为示踪标记物,采用双抗体夹心法检测金黄色葡萄球菌A蛋白,制备了稀土离子标记的免疫层析试纸条,在紫外激发光源下判断反应结果。对该方法的特异性、敏感性和样本检测的适用性进行了分析。结果所制备的试纸条具有良好的特异性,与表皮葡萄球菌、腐生葡萄球菌、单核细胞增生李斯特菌、沙门菌、志贺菌、大肠杆菌、O1/O139群霍乱弧菌、副溶血性弧菌均无交叉反应。用该试纸条检测纯菌液和未经培养的粪便、呕吐物的灵敏度为7.2×104CFU/ml,检测经培养的模拟污染食物样本、粪便样本的检测限最低为7.2×102CFU/ml,反应在15 min内完成。结论本研究成功建立了金黄色葡萄球菌的荧光纳米免疫层析检测方法,能够快速检测食品、奶制品、呕吐物、粪便中的金黄色葡萄球菌,具有良好的特异性和灵敏性,便于开展现场快速检测。     

胶体金免疫层析法检测葡萄球菌B型肠毒素

目的通过优化制备中各关键步骤的实验条件,建立快速检测葡萄球菌B型肠毒素的胶体金免疫层析法。方法采用柠檬酸三钠还原氯金酸法制备直径16nm的胶体金溶液,葡萄球菌B型肠毒素多克隆抗体制备免疫胶体金复合物,组装胶体金免疫层析快速检测试纸条。结果用柠檬酸三钠还原法制备的16nm胶体金溶液呈亮红色,其颗粒大小比较均一。胶体金标记抗体蛋白的最低稳定量为15μg／mL,最适稳定量为18μg／mL,标记的最适pH为7．8。此方法检测限为0．2μg/mL,准确度为94．9％。结论制备了葡萄球菌B型肠毒素胶体金免疫试纸条,该方法简便、快速,而且灵敏度高、稳定性强,适用于食品中葡萄球菌B型肠毒素的快速检测。     

胶体金免疫层析试纸快速灵敏检测沙门菌

目的建立快速、灵敏检测沙门菌的免疫层析试纸方法。方法根据沙门菌的16S DNA序列设计引物进行PCR扩增,并根据PCR产物设计2种特异性探针(探针B和探针F),分别在其5'端和3'端标记Biotin和FAM。PCR产物与探针杂交后应用于免疫层析试纸条。当检测有沙门菌时试纸条会出现2条红线,当没有沙门菌会出现1条红线。对探针的浓度进行优化后,检测沙门菌以及其他7种细菌分析该方法的特异性,并检测不同浓度(107cfu/ml~101cfu/ml)的沙门菌,分析其灵敏度。结果探针的最优浓度为1μmol/L,检测除沙门菌外的其他7种细菌,结果均显示阴性,检测沙门菌的最低浓度为10~2cfu/ml。结论该实验方法的灵敏度高、特异性好,适用于检测沙门菌,有良好的应用性。     

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.     

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.     

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.     

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.     

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.