多重实时荧光PCR快速检测沙门菌和单增李斯特菌

目的 建立可同时检测沙门菌、单增李斯特菌的双重实时荧光PCR快速检测方法,并应用于食品样品的检测.方法 根据GenBank上下载的沙门菌invA基因、单增李斯特菌hlyA基因的保守区序列分别设计特异性引物和TaqMan探针,建立并优化双重实时荧光PCR反应体系.对该方法的特异性、敏感性和稳定性进行评估,并应用于食品标本...     

沙门菌、志贺菌和副溶血性弧菌的多重PCR快速检测技术的建立与应用

目的建立快速同时检测沙门菌、志贺菌和副溶血弧菌的多重聚合酶链反应（PCR）技术。方法根据沙门菌hilA基因、志贺菌ipaH基因及副溶血性弧菌tdh基因序列设计引物，进行PCR扩增及电泳检测。同时优化反应体系，测定特异性和灵敏度。结果该多重PCR技术能在8h内同时检测3种目的菌，通过检测其他9种常见食源性致病菌，结果表明该方法特异性好，多重PCR检测灵敏度分别为：沙门菌10^4cfu／mL，志贺菌10^2cfu／mL，副溶血性弧菌10^4cfu／mL。并进行了人工模拟食品和粪便样品的检测。结论初步建立能在8h内快速、灵敏、特异地同时测定沙门菌、志贺菌和副溶血性弧菌的多重PCR检测技术。     

沙门菌、志贺菌和副溶血性弧菌的多重PCR快速检测技术的建立与应用

目的建立快速同时检测沙门菌、志贺菌和副溶血弧菌的多重聚合酶链反应(PCR)技术。方法根据沙门菌hilA基因、志贺菌ipaH基因及副溶血性弧菌tdh基因序列设计引物,进行PCR扩增及电泳检测。同时优化反应体系,测定特异性和灵敏度。结果该多重PCR技术能在8 h内同时检测3种目的菌,通过检测其他9种常见食源性致病菌,结果表明该方法特异性好,多重PCR检测灵敏度分别为:沙门菌104cfu/mL,志贺菌102cfu/mL,副溶血性弧菌104cfu/mL。并进行了人工模拟食品和粪便样品的检测。结论初步建立能在8h内快速、灵敏、特异地同时测定沙门菌、志贺菌和副溶血性弧菌的多重PCR检测技术。     

实时荧光定量聚合酶链反应检测沙门菌（A～F群）方法的建立和应用

目的 建立实时荧光定量聚合酶链反应(PCR)检测沙门菌,评价其优越性.方法 根据沙门菌保守的HilA基因序列合成引物和探针,建立快速检测沙门菌的实时荧光定量PCR.结果 所建立的实时荧光定量PCR具有简便、快速、敏感性高、特异性强、抗干扰性好和可重复性强等特点,与实际应用检测结果相符.结论 建立的实时荧光定量PCR可用于口岸食品和公共场所从业人员的沙门菌检测.     

实时荧光定量聚合酶链反应检测沙门菌(A～F群)方法的建立和应用

目的建立实时荧光定量聚合酶链反应(PCR)检测沙门菌,评价其优越性。方法根据沙门菌保守的H ilA基因序列合成引物和探针,建立快速检测沙门菌的实时荧光定量PCR。结果所建立的实时荧光定量PCR具有简便、快速、敏感性高、特异性强、抗干扰性好和可重复性强等特点,与实际应用检测结果相符。结论建立的实时荧光定量PCR可用于口岸食品和公共场所从业人员的沙门菌检测。     

基于单基因的实时荧光定量反转录-聚合酶链反应定性检测猪霍乱沙门菌

目的 建立基于单基因的猪霍乱沙门菌的实时荧光定量反转录-聚合酶链反应(rRT-PCR)检测方法。方法 通过对猪霍乱沙门菌基因组及其他血清型沙门菌基因组、人类基因组的生物信息学比对及普通PCR检测分析,获得猪霍乱沙门菌特有而其他细菌及人类基因组中不存在的特异基因SC 1242,针对该基因设计特异性引物,通过优化反应条件,建立检测该靶基因的rRT-PCR方法,利用123株多种血清型沙门菌及非沙门菌菌株RNA评价该方法的特异性及敏感性。同时对猪霍乱沙门菌全血模拟标本进行敏感性检测。结果 利用该方法检测20株猪霍乱沙门菌均为阳性,其余菌株均扩增阴性。对纯菌RNA检测中,rRT-PCR的最低检测限度为50 fg/反应,约为96个拷贝/反应。在以全血模拟样品提取核酸为模板的检测中,敏感性达90 cfu/ml。结论 rRT-PCR方法检测猪霍乱沙门菌特异性好、灵敏度高,对猪霍乱沙门菌的早期诊断具有潜在应用价值。     

基于双启动寡聚核苷酸引物的沙门菌、志贺菌、弯曲杆菌、耶尔森菌多重实时荧光PCR方法的建立

目的建立多重实时荧光PCR同时检测沙门菌、志贺菌、弯曲杆菌、耶尔森菌的方法。方法通过设计针对沙门菌、志贺菌、弯曲杆菌和耶尔森菌的双寡聚核苷酸引物,来建立这4种病原体的多重PCR检测体系。结果建立的多重PCR体系可同时特异性扩增4种病原体的DNA片段。沙门菌和弯曲杆菌的特异性和敏感度都是100.0%,志贺菌和耶尔森菌的敏感度是100.0%,特异性分别是99.3%和98.6%。最低检测限均为103 copies/mL。结论该体系可快速、有效、准确检测出样品中4种病原体的存在,可用于肠胃道相关病原体感染的辅助评价。     

利用实时荧光定量-聚合酶链反应方法检测粪便标本中的伤寒沙门菌

目的 建立针对粪便标本中伤寒沙门菌的检测方法,评价该方法的特异性、敏感性及检测下限,以提高感染人群粪便标本中伤寒沙门菌的检出率。 方法 根据伤寒沙门菌特异基因STY1633设计特异性引物,优化反应条件,建立实时荧光定量-聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,qPCR)反应体系。利用92株常见的非伤寒病原菌及44株伤寒沙门菌的染色体DNA评价该方法的特异性和灵敏性,并对伤寒沙门菌的粪便模拟标本进行检测下限评价,同时利用10份伤寒患者粪便标本及48份其他病原所致发热和/或伴腹泻的患者粪便标本进行特异性、敏感性验证。 结果 利用本方法检测的伤寒沙门菌纯菌及伤寒患者标本均扩增阳性,其余的非伤寒沙门菌、致腹泻的其他5种肠道致病菌及引起发热症状的8种常见非肠道病原菌纯菌及相应患者标本均扩增阴性。在对纯伤寒沙门菌DNA检测中,qPCR法的最低检测限为500 fg/反应,相当于97个拷贝/反应。以粪便模拟样品提取DNA为模板的检测中,增菌前检测下限达104 cfu/g,增菌后可达50 cfu/g。 结论 基于STY1633基因的实时荧光定量PCR方法在检测粪便中的伤寒沙门菌中具有很好的特异性、灵敏度,为伤寒沙门菌的快速诊断及某些不明原因发热及腹泻症状的病原初步鉴定提供了新的简便型手段,对于伤寒的早期诊断及预防控制提供了技术支持。     

多重PCR快速检测3种食源性致病菌

目的建立一种能同时检测金黄色葡萄球菌、产单核李斯特菌和沙门菌3种致病菌的多重PCR检测方法。方法采用LB培养液对金黄色葡萄球菌、产单核李斯特菌和沙门菌标准菌株进行增菌。根据金黄色葡萄球菌的nuc基因、产单核李斯特氏菌的hlvA基因、沙门氏菌的invA基因设计引物,通过多重聚合酶链反应（PCR）对上述3种食源性致病菌的目的基因进行扩增,同时对反应体系进行优化。结果对平均浓度为5cfu／ml的金黄色葡萄球菌、产单核李斯特菌和沙门氏菌在LB培养液中进行8h振荡培养,可以检出阳性结果;把金黄色葡萄球菌、产单核李斯特菌、沙门菌、志贺菌、蜡样芽孢杆菌、大肠埃希菌0157、阪崎肠杆菌7种菌混合在一起提取混合基因组DNA进行PCR扩增,显示出很好的特异性结果。结论建立的多重PCR检测方法适用于金黄色葡萄球菌、产单核李斯特菌和沙门菌的快速检测,具有快速、简便、灵敏的特点,可广泛应用于食品卫生检测、食物中毒应急处理和临床检验等领域。     

三种食源性致病菌多重PCR检测方法的建立及初步应用

目的建立快速检测沙门菌、变形杆菌和金黄色葡萄球菌的多重聚合酶链反应（PCR）方法。方法本研究依据沙门菌侵袭基因正调节蛋白（hilA）基因、变形杆菌溶血素（hpmA）基因和金黄色葡萄球菌特异性pSa-442序列，运用PrimerPremier5．0分别设计3对特异性引物，预计PCR扩增的目的基因片段分别为为580bp、401bp、256bp。通过对单管多重PCR扩增的特异性、敏感性分析以及建立L16（4^3）正交试验对单管多重PCR扩增条件如引物浓度、dNTP浓度和Tm值等的优化，建立了快速同时检测3种食源性致病菌的单管多重PCR方法。结果该方法检测的灵敏度分别为：94．07pg沙门菌DNA，140．85ng变形杆菌DNA，1．41ng金黄色葡萄球菌DNA。模拟检测食品中的混合3种菌，4h培养后样品的最低检测限度分别为：沙门菌10^0菌落形成单位（CFU）／mL、变形杆菌10^1CFU／mL、金黄色葡萄球菌10^0CFU／mL。结论该方法特异性和灵敏度高，检验周期短，可用于对食品中多种致病菌的快速诊检和监控。     

Guns and Knives in New Mexico: Patterns of Penetrating Trauma, 1978–1993

Objective: To identify patterns of nonfatal and fatal penetrating trauma among children and adults in New Mexico using ED and medical examiner data.
Methods: The authors retrospectively sampled in 5-year intervals all victims of penetrating trauma who presented to either the state Level-1 trauma center or the state medical examiner from a 16-year period (1978–1993). Rates of nonfatal and fatal firearm and stabbing injury were compared for children and adults.
Results: Rates of nonfatal injury were similar (firearm, 34.3 per 100,000 person-years; stabbing, 35.1). However, rates of fatal injury were significantly different (firearm, 21.9; stabbing, 2.7; relative risk: 8.2; 95% confidence interval: 5.4, 12.5). From 1978 to 1993, nonfatal injury rates increased for children (p = 0.0043) and adults (p < 0.0001), while fatal penetrating injury remained constant. The increase in nonfatal injury in children resulted from increased firearm injury rates. In adults, both stabbing and firearm nonfatal injury rates increased.
Conclusions: Nonfatal injury data suggest that nonfatal violence has increased; fatal injury data suggest that violent death rates have remained constant. Injury patterns vary by age, mechanism of trauma, and data source. These results suggest that ED and medical examiner data differ and that both are needed to guide injury prevention programs.     

Interprofessional approaches to creating safe,high quality health care

Three supplementary perspectives are presented arguing that interprofessional collaboration is both necessary and desirable. Nonetheless, there are often too many serious intra-professional barriers and obstacles to interprofessional collaboration to make it successful. Some of these barriers, it is argued and illustrated, are found in the multiple ways in which professional identity is tacitly acquired and embodied in the practitioners' habitual, everyday practice. The paper then explores ways in which reflection, especially Second order reflection, can help to elucidate and overcome these obstacles, as well as increasing professional adaptability and competence.     

Prospects for the control of neglected tropical diseases by mass drug administration

The prospects for the control of neglected tropical diseases, including soil-transmitted helminthiasis, shistosomiasis, lymphatic filariasis, onchocerciasis and trachoma, through mass drug administration, are exemplified by the elimination of the trachoma as a public-health problem in Morocco. In spite of this and other striking successes, mass drug administration programs are faced with major challenges resulting from suboptimal coverage and lack of efficacy. At current suboptimal coverage rates, programs may need prolongation for an extended period, increasing costs and undermining sustainability. Community participation through health education and information appears to be crucial to improve coverage and to achieve sustainability. Implementation of complementary measures, such as vector control, improved hygiene and environmental sanitation, are important to further control transmission and to prevent re-emergence of the infection and, again, may only be achieved effectively through community-based initiatives. To reduce costs and to relieve pressure on the health system, combining neglected tropical disease programs in areas where diseases coexist and integration with existing control programs for malaria, tuberculosis and HIV/AIDS is advocated. The risk of developing drug resistance is of particular concern in view of the lack of alternative drugs, and reduced treatment efficacy due to emerging resistance is evident for the soil-transmitted helminths and onchocerciasis. Given the risk for the development of drug resistance and the need for a high degree of participation, close attention should be paid to the monitoring of the coverage and efficacy of the different program components.     

Bacterial meningitis: an update of new treatment options

The outcome of bacterial meningitis critically depends on the rapid initiation of bactericidal antibiotic therapy and adequate management of septic shock. In community-acquired meningitis, the choice of an optimum initial empirical antibiotic regimen depends on the regional resistance patterns. Pathogens resistant to antibacterials prevail in nosocomial bacterial meningitis. Dexamethasone is recommended as adjunctive therapy for community-acquired meningitis in developed countries. In comatose patients, aggressive measures to lower intracranial pressure <20 mmHg (in particular, external ventriculostomy, osmotherapy and temporary hyperventilation) were effective in a case–control study. Although many experimental approaches were protective in animal models, none of them has been proven effective in patients. Antibiotics, which are bactericidal but do not lyse bacteria, and inhibitors of matrix metalloproteinases or complement factor C5 appear the most promising therapeutic options. At present, vaccination is the most efficient method to reduce disease burden. Palmitoylethanolamide appears promising to enhance the resistance of the brain to infections.     

Imaging choices in occult hip fracture

Background: Hip fracture is a common injury, with an incidence rate of > 250,000 per year in the United States. Diagnosis is particularly important due to the high dependence on the integrity of the hip in the daily life of most people. Objectives: In this article we review the literature focused on hip fracture detection and discuss advantages and limitations of each major imaging modality. Discussion: Plain radiographs are usually sufficient for diagnosis as they are at least 90% sensitive for hip fracture. However, in the 3–4% of Emergency Department (ED) patients having hip X-ray studies who harbor an occult hip fracture, the Emergency Physician must choose among several methods, each with intrinsic limitations, for further evaluation. These methods include computed tomography, scintigraphy, and magnetic resonance imaging. Conclusion: We present an evidence-based algorithm for the evaluation of a patient suspected to have an occult hip fracture in the ED. Also outlined are future directions for research to distinguish more effective techniques for identifying occult hip fractures.     

Systematic Reviews Published in the April 2012 Issue of the Cochrane Library

ABSTRACT

The Cochrane Library of Systematic Reviews is published quarterly as a DVD but monthly online. The April 2012 issue (second DVD for 2012) contains 5045 complete reviews, 2182 protocols for reviews in production, and 17,084 short summaries of systematic reviews published in the general medical literature. In addition, there are citations of 674,000 randomized controlled trials, and 15,400 cited papers in the Cochrane methodology register. The health technology assessment database contains just over 11,000 citations. One hundred and seventeen new reviews have been published in the last 3 months of which 12 have potential relevance for practitioners in pain and palliative medicine. The impact factor of the Cochrane Library stands at 6.186. Readers are encouraged to access the full report for any articles of interest as only a brief commentary is provided.     

Platelets,thrombosis, coagulation,and atherosclerosis

When I first got the invitation to join a medical delegation going to Moldova, I thought for a moment that our destination was the fictional country in the old Marx Brothers movie Duck Soup. On further checking, it turns out that entertaining place was called Freedonia. I now know that Moldova is indeed a real country, bordered on the west by Romania and on the other three sides by the Ukraine. It is a proud country, rich with traditions, and its people are warm, giving, eager to learn ways to improve their healthcare system, and deeply appreciative of our attempts to help them in the task.     

Advance Care Planning

Predictors of patient wishes and influence of family and clinicians are discussed. Research findings on patient decision-making relating to preferences in end-of-life care are described. Advance directives and durable powers of attorney are defined and differentiated. Most patients have not participated in advance care planning and the need for more effective planning is documented. Appropriate times for discussions of such planning are described. Scenarios discussed include terminal cancer, chronic obstructive pulmonary disease, AIDS, stroke, and dementia. Patient satisfaction is discussed, as is a structured process for discussions about patient preferences. Results of patient responses to hypothetical scenarios are described. Invasiveness of interventions, prognosis and other factors that favor or discourage patient preferences for treatment are discussed. Findings resulting from research funded by the Agency for Healthcare Research and Quality (AHRQ) are discussed. This research can help providers offer end-of-life care based on preferences held by the majority of patients under similar circumstances.     

Spiritual Dimensions of Nursing Practice. edited by V. B. Carson

The Cochrane Library of Systematic Reviewsis published quarterly. Issue one for 2004 of the library was published in February 2004. This issue contains 3,329 reviews and protocols of which 1,921 are fully published reviews. The trials database now stands at over 400,000 records with an additional 4,427 one-page summaries of non-Cochrane reviews in the NHS database of reviews of effectiveness (DARE). This version of the library contains the results of an extensive search for RCTs on EMBASE. The latest library contains 84 new reviews, seven are considered relevant to practitioners in pain and palliative care. References are published in the same format as the citation for Cochrane reviews.     

Évaluation,par l’imagerie ultrasonore,du recrutement musculaire,de la force motrice et de la détente verticale du quadriceps après électromyostimulation (cas de 15 étudiants de l’INJS)

The increase of strength seems to be an element of performance in some sports.