荧光PCR熔解曲线法检测结核分枝杆菌耐药性价值及耐药特征分析

目的 评价荧光PCR熔解曲线法(熔解曲线法)对结核分枝杆菌(MTB)耐药性检测价值。方法 采用熔解曲线法与传统液体药敏法同时检测156例临床分离培养MTB对抗结核药物利福平(RFP)、异烟肼(INH)、乙胺丁醇(EMB)、链霉素(SM)和氟喹诺酮类药物耐药情况,并对2种检测方案结果进行比较。结果 与液体药敏检测相比,熔解曲线法对5种抗结核药物耐药性检测结果差异无统计学意义(P> 0.05),以液体药敏检测为金标准,熔解曲线法检测RFP、INH、EMB、SM以及氟喹诺酮类药物耐药性的灵敏度分别为100.00%、75.86%、61.54%、75.00%、90.00%,特异度分别为97.10%、95.28%、99.30%、96.21%、100.00%,Kappa值分别为0.885、0.721、0.707、0.724、0.944,表明2种检测方法一致性好。结论 荧光PCR熔解曲线法对抗结核药物RFP、INH、EMB、SM及氟喹诺酮类的耐药性与液体药敏法检测效果一致,灵敏度、特异度均较高,具有较好的临床应用价值。     

分格平板分枝杆菌药敏培养基在结核分枝杆菌药敏检测中的应用

目的 评价分格平板分枝杆菌药敏培养基,在结核分枝杆菌药敏检测中的应用.方法 收集临床分离的结核分枝杆菌51株,药敏方法分别采用分格平板分枝杆菌药敏培养基比例法和罗氏培养基比例法(试管比例法).结果 试管比例法:51株结核分枝杆菌的耐药率对异烟肼为68.6％、利福平为51.0％、乙胺丁醇为49.0％、链霉素为52.9％;与试管比例法相比较,分格平板琼脂比例法药敏结果2株对异烟肼耐药、1株利福平耐药和2株乙胺丁醇耐药菌株与其不符,其余结果一致;4种一线抗结核药物的药敏结果符合率分别为异烟肼94.3％、利福平96.3％、乙胺丁醇92.0％、链霉素100.0％;51株分离的临床结核分枝杆菌中泛耐药24株,多药耐药7株,而对4种药物全敏感12株,敏感率仅为23.9％.结论 分格平板琼脂比例法操作简便,结果符合率高,容易观察,能广泛应用各级临床结核实验室.     

显微镜观察药物敏感性试验检测结核杆菌的耐药性

目的 探讨显微镜观察药物敏感性试验(MODS)同时快速检测结核杆菌对4种一线抗结核药物耐药性的效果.方法 用24孔细胞培养板进行结核分枝杆菌(MTB)液体药敏检测,并对最佳检测条件进行探讨;将建立的MODS技术用于测定66株MTB临床分离株对链霉素、异烟肼、利福平、乙胺丁醇的耐药性,并与传统罗氏药敏结果 进行比较,对两法检测结果 不符的菌株进行最低抑菌浓度(MIC)测定.结果 MODS检测链霉素、异烟肼、利福平、乙胺丁醇药敏结果 与罗氏药敏结果 符合率分别为97.0%、90.9%、95.5%、86.4%.如以传统罗氏药敏结果 为判断标准,则MODS检测链霉素的敏感度为96.0%、特异度为97.6%、阳性和阴性预测值(PPV、NPV)及准确性分别为96.0%、97.6%和97.0%;检测异烟肼、利福平和乙胺丁醇的敏感度、特异度、PPV、NPV及准确性分别为100%、85.4%、81.0%、100%、90.9%;96.2%、95.0%、92.6%、97.4%、95.5%和73.7%、91.5%、77.8%、89.6%、86.4%.共有16株菌的20个药敏结果 MODS与传统罗氏药敏结果 不符,MIC检测结果 有14株16个药敏结果 与MODS结果 一致,4株菌4个药敏结果 与罗氏药敏结果 一致.结论 MODS同时检测链霉素、异烟肼、利福平、乙胺丁醇药敏结果 具有快速、操作简便、价廉等优点,与传统罗氏药敏结果 有较高的符合率,可作为MTB耐药性的快速检测方法 之一.     

结核分枝杆菌耐多药性基因芯片联合检测

目的利用基因芯片技术检测结核分枝杆菌(MTB)对链霉素(SM)、乙胺丁醇(EMB)、利福平(RFP)和异烟肼(INH)等4种抗生素的耐药性。方法应用基因芯片法联合检测300株MTB临床分离株的SM、EMB、RFP和INH耐药性,并与BACTEC960法检测结果进行比较,将结果不相符的菌株进行测序,并比较结果。结果基因芯片法测定300株MTB临床分离株,SM敏感188株、耐药112株,以rpsL43、88位aag突变为agg为主;EMB敏感251株,耐药49株,以embB 306位atg突变为gtg为主;RFP敏感211株,耐药89株,以ropB 516位aag突变为gag为主;INH敏感210株,90株,以KatG 315位gtg突变为gag为主;基因芯片法检测SM、EMB、RFP和INH耐药的敏感性、特异性分别为81.1%,99%;60.0%,100%;72.0%,98%;75.0%,99%;2种测定方法结果不相符菌株中,基因测序结果显示,基因芯片法检测点突变具有高度特异性。结论基因芯片法联合检测MTB对SM、EMB、RFP和INH的耐药性,快速且特异性高,可作为MTB临床检测耐药性的快速筛选方法。     

Gene-Xpert MTB/RiF与熔解曲线法检测结核分枝杆菌利福平耐药分析

目的评价Gene-Xpert MTB/RiF法与熔解曲线法检测结核分枝杆菌利福平耐药的不同应用价值。方法入选235例涂阳结核病患者痰标本分别做传统比例法药敏试验、Gene-Xpert MTB/RiF和熔解曲线法检测结核分枝杆菌的利福平耐药,以传统药敏试验作为金标准进行比较。结果 235例标本中比例法药敏试验利福平耐药12例(5.1%)、Gene-Xpert MTB/RiF利福平耐药例15例(6.4%)、熔解曲线法利福平耐药19例(8.1%),Gene-Xpert MTB/RiF和熔解曲线法敏感度分别为91.7%和100.0%;特异度分别为98.2%和96.7%。结论Gene-Xpert MTB/RiF和熔解曲线法两者均可应用于结核分枝杆菌利福平耐药性的快速检测,Gene-Xpert MTB/RiF法在我国开展结核菌耐多药筛查具更有较强的实用性,有更广的应用前景。     

rpoB基因突变与结核分枝杆菌利福平耐药相关性研究

目的 探讨临床患者痰标本中分离的结核分枝杆菌rpoB基因突变与利福平耐药之间的关系. 方法 对115株结核分枝杆菌应用L-J药敏培养法,检测对利福平、异烟肼、乙胺丁醇、链霉素、氧氟沙星的耐药情况;根据rpoB基因序列设计包含利福平耐药决定区（RRDR）的扩增引物,PCR法进行扩增,运用生物信息学方法对耐药决定区扩增产物序列进行比对分析,对耐药株与非耐药株基因突变率进行统计学分析. 结果 115株结核分枝杆菌经比例法药敏试验,RIF耐药株53株,占所有菌株的46.09％;53株RIF耐药株中39株rpoB基因存在不同位点的变异,突变率为73.58％,突变位点包括531、526、522、516、513、511,最主要的突变位点集中在531位,引起氨基酸呈Ser531Gln、Ser531Leu、Ser531Trp 3种形式改变;RIF敏感株62株,其中有2株检测到rpoB基因的突变,敏感株与耐药株突变率经McNemar统计检验,差异有统计学意义（P＜0.05）. 结论 岳阳地区rpoB基因变异与利福平耐药呈高度相关,rpoB基因最主要的突变位点位于531位和526位.     

反向斑点杂交技术在结核分枝杆菌耐药性检测中的应用研究

目的利用反向斑点杂交技术，建立结核分枝杆菌对利福平（RFP）、异烟肼（INH）、链霉素（SM）和乙胺丁醇（EMB）耐药相关基因突变的快速检测方法。方法根据结核分枝杆菌标准株H37Rv序列，自行设计针对rpoB、katG、inhA、rpsL和embB基因中常见耐药突变区的系列寡核苷酸探针，制成膜芯片，并对64例结核分枝杆菌临床株的基因突变情况进行检测。结果在36株RFP耐药菌中，有32株被检出在rpoB基因上发生突变，突变检出率为88．89％（32／36）；在39株INH耐药菌中，有30株被检出在katG或inhA基因上发生基因突变或缺失，检出率为76．92％（30／39）；在34株SM耐药菌中，有25株被检出在rpsL基因分析部位发生突变，检出率为73．5％（25／34）；在32株EMB耐’药菌中，有19株在embB基因分析部位发生突变，检出率为59．4％（19／32）。膜芯片检出的突变与基因测序结果一致。结论用膜芯片检测结核分枝杆菌对利福平（RFP）、异烟肼（INH）、链霉素（SM）和乙胺丁醇（EMB）四种主要一线抗结核药的耐药性，具有较高的特异性和敏感性，可作为常规药敏方法的补充，用于指导开展早期、有效的临床化疗。     

荧光PCR探针熔解曲线法检测结核分枝杆菌耐利福平突变研究

目的对荧光PCR熔解曲线法检测结核分枝杆菌耐利福平突变方法进行临床研究,评价检测能力及在国境口岸的应用价值。方法应用荧光PCR熔解曲线法检测结核病人临床分离结核分枝杆菌,以传统药物敏感性试验为标准,获得该方法的灵敏性、特异性。结果对347例结核病人临床分离培养样本,传统药物敏感性试验检出271例利福平敏感标本,76例利福平耐药标本。荧光PCR熔解曲线法检出269例利福平敏感标本,78例利福平耐药标本,灵敏性为93.42%,特异性为97.42%,符合率为96.54%。结论荧光PCR熔解曲线法检测速度快速、灵敏度高、特异性强,可用于结核分枝杆菌利福平耐药突变的快速检测,适于国境口岸对耐多药结核病的快速筛查。     

荧光PCR熔解曲线法检测结核分枝杆菌耐利福平突变研究

目的 对荧光PCR熔解曲线法检测结核分枝杆菌耐利福平突变方法进行临床研究,评价其检测效能及在国境口岸应用价值.方法 应用荧光PCR熔解曲线法检测结核病人临床分离标本,与传统药物敏感性试验对比,衡量该方法的灵敏度、特异性.结果 对347例结核病人临床分离培养样本,药敏试验检出271例利福平敏感标本,76例利福平耐药标本;...     

广泛耐药结核快速检测方法的比较研究

目的应用改良显微镜观察药物敏感性检测法(MODS)对临床结核分离株进行检测,完善和规范对广泛耐药结核(XDR-TB)快速检测的方法,评价其临床应用价值。方法对已做过罗氏比例法药敏的临床分离株进行MODS、BECTEC MGIT 960法药敏试验,将3种药敏结果对比分析,评估MODS检测XDR-TB的应用性价值。结果 29株MTB临床分离株,应用MODS技术检测出20株多耐药结核(multi drug-resistant tuberculosis,MDR-TB),其中XDRTB有12株。MODS检测利福平、异烟肼、链霉素和乙胺丁醇的药敏结果与罗氏比例法的符合率分别为93.1%、100.0%、79.3%、75.9%,与BECTEC MGIT 960法的符合率分别为100.0%、100.0%、100.0%、58.6%。17株MTB临床分离株,MODS检测左氧氟沙星的药敏结果与罗氏比例法的符合率为94.1%。结论 MODS检测利福平、异烟肼的药敏结果与罗氏比例法、BECTEC MGIT 960法的符合率高,检测左氧氟沙星的药敏结果与比例法的符合率较高,具有快速、价廉、操作简便的优点,可作为临床检出MDR-TB、XDR-TB的快速检测方法之一。     

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.     

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.