微型寡核苷酸芯片检测脑脊液病原菌研究与应用

目的建立快速检测脑脊液标本中常见病原菌的微型寡核苷酸芯片。方法根据脑脊液中常见病原菌16S rRNA基因序列设计通用引物和检测探针;探针固定于尼龙膜上制成寡核苷酸芯片;通用引物扩增病原菌DNA并标记生物素,产物与芯片进行反向杂交检测;对该体系的敏感性和特异性进行了测试,并检测了32例临床病原菌感染的脑脊液标本。结果所有实验菌株均只与芯片上相应探针杂交。该法最低可检测出10 CFU/ml的大肠埃希菌;32例临床本的检测结果与常规鉴定法完全一致。结论该寡核苷酸芯片法能够准确、敏感、快速地检测脑脊液标本中的病原菌。     

荧光原位杂交法对血培养标本中细菌和真菌鉴定效果评价

目的建立一种阳性血培养标本的检测和鉴定的快速方法. 方法采用荧光原位杂交法,Bact/Alert仪器报警后,取阳性血培养液15μl于玻片上,全细胞杂交1h,同时将阳性标本转种培养基平板,用VITEK 仪器法鉴定,比较杂交法与仪器法的符合率. 结果杂交法与仪器法测定了118份临床阳性标本,葡萄球菌属的符合率96.3%,链球菌属和肠球菌属的符合率100%,革兰阴性杆菌的符合率86.8%,白色念珠菌的符合率100%,总符合率91.5%. 结论联合应用Bact/Alert血培养仪及荧光原位杂交法, 能显著缩短阳性血培养标本中的细菌和真菌的鉴定时间.     

rRNA扩增直接检测艾滋和结核共感染的结核分枝杆菌的初步研究

目的 建立一种快速、准确、简便的检测结核分枝杆菌的方法.方法 应用间接转录扩增技术,以结核分枝杆菌特异性rRNA为扩增目标,扩增片段被特异性DNA探针结合（杂交）,再经杂交保护分析法筛选,最后以化学发光仪检测被标记的rRNA.用该法检测62例临床标本.结果 62 例艾滋合并结核患者痰标本检出阳性9例.结论 rRNA扩增直接检测结核分枝杆菌方法作为结核病诊断的一项新的分子生物学检测方法,具有重要的临床应用价值.     

实时荧光定量PCR检测痰标本中军团菌属

目的建立荧光定量PCR方法,用于检测军团菌属特异性16S rRNA基因,并探讨广州地区军团菌属感染状况。方法利用军团菌属特异性16S rRNA基因保守序列设计引物和探针,优化反应条件和反应体系,对嗜肺军团菌、非嗜肺军团菌及其他细菌进行检测,验证该方法的特异性、敏感性、重复性,对广东省中医院采集的532例肺部感染患者痰液标本进行检测。结果该方法检测军团菌属所有标准菌株均出现阳性信号,其他非军团菌属细菌检测结果均为阴性;检测灵敏度为102CFU/ml,临床检测532例肺部感染患者的痰液标本,荧光法检出军团菌阳性43例,阳性率为8.08%,16S rRNA PCR法加基因测序验证阳性率为7.71%,两者检测结果差异无统计学意义,表明其具有较好的符合性和等效性。结论 16S rRNA基因荧光定量PCR法检测患者痰液标本中军团菌属,具有快速、敏感、特异等特点,适用于临床军团菌属感染调查及快速检测。     

采用通用引物PCR与机器码识别技术快速鉴定血液感染病原菌

目的 建立一种快速检测血液感染病原菌的方法.方法 选取7种常见血液感染病原菌,在其16S rRNA末端和23S rRNA始端的保守区设计通用引物,对16S-23S rRNA基因间区进行扩增,然后将PCR产物的电泳图谱进行机器码识别,以鉴定病原菌.结果 通用引物能扩增出7种病原菌,且每种菌具有不同片段长度(bp):大肠埃希菌(716、808),金黄色葡萄球菌(732、827),铜绿假单胞菌(833),表皮葡萄球菌(630、722、817),肺炎链球菌(606),肺炎克雷伯菌(550、705、797),粪肠球菌(591、693);此图谱可经机器码识别,对各种病原菌进行快速鉴定.结论 采用通用引物PCR和机器码识别技术可简便、快速、特异地鉴定常见血液感染病原菌.     

酶显色法基因芯片检测幽门螺杆菌克拉霉素耐药基因

目的 建立酶显色法基冈芯片快速、准确检测幽门螺杆菌(Hp)克拉霉素耐药基因.方法 设计-对引物,以地高辛标记其中一条引物,扩增Hp 23S rRNA部分基因,得到285 bp DNA片段.根据23S rRNA上的5个位点设计探针和制作芯片,扩增产物与芯片杂交,酶显色后扫描判读结果.结果 63例Hp阳性标本中,芯片结果显示突变率:A2143G为3.17%、T2182C为11.11%、A2143G+T2182C为14.29%;酶显色法芯片和DNA直接测序法检测Hp 23S rRNA基因多态性结果符合率为96.83%;质粒浓度为1.53×10~2拷贝/μl时芯片法仍可检测到野生和突变信号.结论 酶显色法芯片检测实现了高通量的同时具有较高敏感性和特异性,能同时检测23S rRNA的多种突变型,无需特殊仪器,可以用于指导临床合理用药和进行流行病学调查,有一定的临床应用价值.     

运用23S rDNA芯片技术进行食源性感染常见致病菌的快速鉴定

[目的]应用23S rDNA芯片技术，建立食源性感染常见致病菌的快速鉴定方法。[方法]利用带正电荷尼龙膜为芯片载体，合成寡核苷酸探针，点样于载体制成寡核苷酸芯片，对食源性感染常见致病菌23S rDNA基因片段扩增产物进行杂交检测。[结果]获得扩增食源性感染常见致病菌23S rDNA基因片段的特异性引物，并在同一条件下扩增出14种细菌目标片段。在均一的杂交条件下与寡核苷酸芯片杂交，用地高辛酶联免疫法进行显色。结果表明，10种细菌杂交结果显示很好的灵敏度和特异性，4种细菌由于探针因素或杂交条件不合适未能显示预期的种(属)杂交结果。对于食源性感染模拟标本，本方法也可以获得预期的结果。[结论]建立的23S rDNA芯片技术在食源性感染常见致病菌鉴定上快速准确的优点，为食源性感染的诊治与预防提供了有效的技术手段和方法依据。     

实时荧光定量PCR技术在群体感染性腹泻病原体快速筛查中的应用

目的:建立快速筛查感染性腹泻标本的实时荧光定量PCR法,考察其在群体感染性腹泻病原体快速筛查中的应用.方法:采用国家标准的细菌培养法(或传统ELISA法)对2006年丽水地区发生的总计14起共487例群体感染性腹泻标本进行病原体筛查,同时采用实时荧光定量PCR方法对该487例标本进行检测,比较两者的实验结果,并考察实时荧光定量PCR方法的特异性和灵敏度.结果:实时荧光定量PCR检测结果与传统检测方法的符合率为97.54%,除完全符合的278例标本之外,另有7例采用传统方法检测为阴性的标本用实时荧光定量PCR检测显示为阳性.表明实时荧光定量PCR方法具有更高的灵敏度;特异性和灵敏度实验表明实时荧光定量PCR方法的特异性达100%,灵敏度为102拷贝/ml.结论:实时荧光定量PCR法具有准确性好、灵敏度高、检测快速等优点.是一种理想的群体感染性腹泻病原体快速筛查的方法.     

聚合酶链反应在检测脑脊液病原菌中的应用

目的 应用聚合酶链反应(PCR)建立快速检查方法来检测脑脊液中是否含有病原菌.方法 对138例脑脊液标本,以病原菌16S rRNA基因为靶序列,采用1对通用引物对其进行PCR扩增检测,并同时作细菌培养.结果 138例脑脊液标本中,PCR方法检测出55例,阳性率为39.86%,培养法检出24例,阳性率为17.39%,PCR方法检出率明显高于培养法(P<0.01).结论 该方法具有快速、敏感、特异、准确等优点,是一种可靠的实验手段.     

基于16S和23S rDNA基因芯片检测和鉴定七种临床常见病原菌

目的 以细菌16S rDNA和23S rDNA基因为靶序列建立可检测临床七种常见病原菌寡核苷酸芯片系统.方法 采用双重PCR扩增标本中靶细菌16S和23S rDNA基因片段.构建能同时检测肠出血性大肠埃希菌O157:H7、副溶血性弧菌、沙门菌、霍乱弧菌、单核细胞增生李斯特菌、空肠弯曲菌和志贺菌的寡核苷酸芯片,并考核芯片的检测特异性、灵敏度和重复性.采用所建立的寡核苷酸芯片检测81例腹泻患者粪便样本.结果 双重PCR可同时扩增上述七种病原菌的16S和23SrDNA基因靶序列.所研制的寡核苷酸芯片检测灵敏度可达103 cfu/ml,非靶细菌无阳性结果 ,不同批间和批内芯片的变异系数为3.89%～5.81%.寡核苷酸芯片检测粪便样本的阳性率为39.5%(32/81),与常规细菌学检查法检测结果 的符合率达到96.3%(78/81),菌种鉴定结果 符合率为96.8%(31132).结论 研究建立的寡核苷酸芯片法在检测七种病原菌时具有简便、快速、准确、高通量等优点,适合于临床样本检测及流行病学现场调查.     

Trends in meningococcal disease in Italy in 1988

Meningococcal disease in Italy decreased 15% in 1988 from the previous year (290 vs. 342 cases). The decline was particularly evident in military cases (1.7/100,000 in 1988 vs. 5/100,000 in 1987) reflecting the full coverage of bivalent serogroup (A + C) meningococcal polysaccaride vaccine in army recruits, achieved since January 1988. The highest proportion of cases was seen in people older than 25 years of age (25%). Serogroup C constituted 60% of the isolates, while 19% belonged to serogroup B. The proportion of strains resistant to sulphonamides was 45%, while 15% were resistant to Minocycline and none to Rifampin. Out of the five military cases, only one (due to serogroup C) was attributable to the vaccine failure. A single coprimary case, but no secondary cases occurred among civilians. These findings are consistent with the trends reported in Italy in the previous years.     

Benzene in blood and phenol in urine in monitoring benzene exposure in industry

Determinations of benzene concentration in blood and of phenol in urine were made by head-space gas chromatography techniques on samples taken near the end of the work day from two groups of workers potentially exposed to low levels of benzene in the work-place atmosphere. Preliminary results suggest that benzene in blood is more reliable than phenol tests for assessing both exposure and uptake of benzene. Normal values of phenol in urine (10 mg/liter or less) were found in nearly all those cases in which benzene was detected in the blood.     

2015年郑州市饮用水放射性水平状况调查

目的了解郑州市不同地区、不同季节、不同水源类型的放射性水平。方法分别于丰水期和枯水期采集郑州市区黄河水源水、井水源水、丹江口水源水、出厂水和末梢水各一份;以县为单位,每单位采集出厂水、末梢水、水库水、河水、井水各1份,按《生活饮用水生活标准检验方法》(GB/T 5750.13-2006)检测饮用水中总α和总β放射性水平,依据《生活饮用水卫生标准》(GB 5749-2006)进行判定。结果丰水期、枯水期水体总α、总β放射性水平均低于国家标准限值。结论郑州市不同水源类型均处于正常的天然放射性本底水平。     

Occupational stress in nurses in psychiatric institutions in Taiwan

Nurses are known to be exposed to occupational stress. However, occupational stress is not well documented for nurses in psychiatric institutions in Taiwan. A cross-sectional study was conducted to explore the work-related stress and risk factors of nurses in psychiatric institutions in Taiwan. A structured questionnaire was distributed to nurses at five state-owned psychiatric hospitals in Taiwan in 2001. Demographic information, working environment, and personal health status were inquired. Occupational stress was assessed based on the Chinese version of Job Content Questionnaire (JCQ). General health status and mental health were evaluated by the International Quality of Life Assessment Short Form-36 (IQOLA SF-36). A total of 573 questionnaires were disseminated to nurses and 518 (90.4%) were satisfactorily completed by nurses, including 408 female full-time nurses who had been in their current work for more than 6 months. In the past one month, 17.2% of nurses reported being under significant stress often or always. Assault episodes were reported by 45.1% of nurses in the past 6 months. Among the nurses, 16.9%, 25.2%, 50.0%, and 7.8% belong to the "High strain", "Low strain", "Active", and "Passive" groups, respectively. Perceived occupational stress was associated with young age, widowed/divorced/separated marital status, high psychological demand, low workplace support, and threat of assault at work. Lower general health score was associated with low job control, high psychological demand, and perceived occupational stress. A lower mental health score was associated with low job control, high psychological demand, low workplace support, and perceived occupational stress. We concluded that nurses in psychiatric institutions are under significant stress related to work factors.     

Changes in nursing administration in supporting transplantation in Brazil

This historical and bibliographic study aimed to understand how Nursing was organized to support care in transplantation. The HISA, LILACS, BDENF, PERIENF and DEDALUS databases were consulted, and thirteen references were found, ten of which were scientific articles, two were master's dissertations and one was a doctoral thesis. The span of time chosen for study ranges from the date of the first kidney transplant in Brazil (1965), to the date of publication of the last scientific article found in the databases mentioned above (2003). After reading these articles, the ones that were similar in topic were grouped together, thus creating the thematic axis for the presentation of the results. The results showed that the Nursing profession has played an important and active role in transplants ever since the first procedure in 1965.     

2019年中国肿瘤流行病学研究进展

恶性肿瘤已成为中国居民的主要死因之一。近些年来,中国政府积极推进肿瘤预防和控制领域的研究工作,取得了很大的进步。本文简要汇总中国在肿瘤流行病学领域的研究进展,具体包括2019年的肿瘤负担、癌症危险因素及其干预、筛查和早期发现、癌症防治专项行动(2019-2022年)方案等方面,以期为我国肿瘤防治工作的有效开展提供技术支撑和理论依据。     

2012年绵阳市蔬菜中农药残留状况

目的 了解绵阳市2012年市售蔬菜中农药残留情况,为食用蔬菜监管提供依据.方法 全部样品均按照GB/T 5009-2003的方法进行农药的残留检测.依据GB 2763-2005《食品中农药最大残留限量》进行判定.结果 2012年共检测蔬菜165份,农药检出率和超标率分别为72.12％和23.64％.豆类、根茎类、叶菜类、瓜果菜类和食用菌类蔬菜检测农药残留超标率分别为8.00％、23.53％、36.00％、5.26％和27.03％,差异有统计学意义(x2=11.41,P＜0.01).不同种类农药超标率差异有统计学意义(x2 =62.42,P＜0.01).夏季和冬季蔬菜农药超标率差异无统计学意义(x2=3.44,P＞0.05).结论 绵阳市叶菜类蔬菜中农药残留情况较为严重,应采取有效措施,加强蔬菜的监督管理,从源头禁止高毒农药的使用,加大蔬菜种植、销售环节的监测,确保市民食用蔬菜的安全.