新冠病毒核酸检测试剂盒的性能分析与评价

目的：对4种新型冠状病毒核酸检测试剂盒的性能包括敏感性与特异性、最低检出限、精密度和分析特异性进行评价,为实验室选择新冠病毒核酸检测试剂盒提供参考,促进新冠肺炎病毒核酸检测技术的创新改进。方法：以20例新冠病毒确诊阳性和10例确诊阴性的咽拭子作为研究样本,经提取得到RNA后根据4种新冠病毒核酸检测试剂盒说明书对其进行检测,分析扩增结果,评估各试剂盒的差异。结果：检测结果显示4种试剂盒对30例研究样本的阴阳性判定情况不尽相同,其中a试剂盒的敏感性和特异性最高,均达到100%;3例阳性样本稀释到100倍时,c试剂盒只能检出其中的1例,而其他三种试剂盒能检出其中的2例,表明c试剂盒的检测敏感性低于其他三种;4种试剂盒在精密度实验中检测样本得到的Ct值的CV值都较接近,且都<10%;加入了8种常见的高浓度干扰物质的临界阳性样本经4种试剂盒检测,结果依然为弱阳性,说明无干扰;4种试剂盒检测5例含非目标病毒的临床样本,检测结果为阴性,即未发生交叉反应。结论：新冠病毒核酸检测试剂盒对低浓度核酸样本的检测能力有所差异,各试剂盒不同的检出限也造成了总体上的敏感性和特异性的差异,实验室在选择新冠病毒...     

B族链球菌核酸检测试剂盒临床应用对比评价

目的 通过对比评价不同厂家B族链球菌（group B streptococcus,GBS）核酸检测试剂盒的临床和实验室应用情况,为B族链球菌的临床检测提供参考。方法 利用A和B两个厂家的B族链球菌核酸检测试剂盒平行检测964例临床收集样本中的GBS DNA,同时检测一例临床高值样本梯度稀释后的系列样本;另外利用A、B、C三个厂家对GBS培养菌株进行梯度稀释检测,统计并对比检测结果。结果 A、B两个厂家核酸检测试剂盒阳性符合率100.0%,阴性符合率99.3%,总符合率99.7%,Kappa值为0.994,结果一致性高;两个厂家试剂盒同时检测来源于同一例、不同倍数稀释后的临床样本,结果显示A厂家试剂盒检测灵敏度要高于B厂家试剂盒约两个数量级。对GBS培养菌株进行检测对比,结果显示A厂家试剂盒的检测灵敏度高于或相同于B厂家试剂盒,C厂家试剂盒灵敏度次之。结论 A厂家的B族链球菌核酸检测试剂盒应用于临床GBS的检测,具有方便快捷、灵敏度高的特点。     

浙江省血站核酸集中化检测质量评价体系的建立

血液安全一直是世界卫生组织关注的焦点,世界范围内由输血导致感染的人数达到百万人,5%~10%的新发HIV感染是通过输注窗口期血液引起的。HBV、HCV、HIV是经血传播的主要病原体,血清学检测由于窗口期长、免疫逃逸、免疫静默、隐匿性感染等因素造成漏检,血液安全得不到有效保障。核酸扩增检测技术(NAT)有效弥补血清学检测的不足.     

核酸扩增(PCR)荧光定量检测方法在检测细菌性食物中毒病原菌中的应用

在食物中毒事件中细菌性食物中毒传统的检测方法是增菌培养、分离培养、纯培养、形态及血清型鉴定、生化鉴定等，做出初步鉴定需要48h以上，本文采用核酸扩增（PCR）荧光定量检测技术。检测细菌性食物中毒标本中的病原微生物，结果2h内得出检测结论，为进一步救治和提出防范措施争取了时间。现将检测方法介绍如下。     

新型冠状病毒核酸检测分析

在新型冠状病毒具备较高的传染性和隐蔽性,在发作前会经历14天的潜伏期,新型冠状病毒的核酸检测中值得注意的是相当一部分疑似病例都是通过多次检测才确诊了结果,阳性患者的核酸检测阳性率不到五成,检测反馈效率不高,虽然检测仍然是有效途径,但这无疑对于检测的结果、效率和公共卫生安全问题有所影响,需要加以重视和改善。因此,本文对新型冠状病毒核酸检测进行简要分析,主要从假阴性、假阳性的引起因素出发去分析检测问题,并通过一定的实验结果对比来观察核酸检测问题,以期能够为医疗机构尤其是基层医院提供一些核酸检测工作上的帮助。     

为什么确诊新冠肺炎需要核酸检测试剂盒?

病毒主要由遗传物质和蛋白质组成,病毒的遗传物质就是由一个或多个核酸分子(DNA或RNA)组成的基因组。新型冠状病毒(2019-nCoV)是一种RNA(核糖核酸)病毒,其遗传物质是单链RNA,这是新型冠状病毒最核心、最准确的标志。核酸检测试剂盒是能够直接检测病毒核酸的工具。     

某市基地实验室新型冠状病毒核酸检测系统的性能评价

目的 评价某市核酸检测基地新型冠状病毒核酸检测系统的性能,以保证核酸检测结果的可靠性。方法 采用强阳性、弱阳性性能验证参考品及临床阴性样本对核酸检测系统的符合率、精密度、检测限和特异性进行验证。结果 符合率验证实验中,5份不同浓度阳性参考品检测结果均为阳性,5份阴性样本检测结果均为阴性,总符合率为100%;精密度验证实验中,强阳性和弱阳性参考品的ORF1ab基因批内精密度CV值分别为0.43%和1.31%,批间精密度CV值分别为0.56%和1.40%,强阳性和弱阳性参考品的N基因批内精密度CV值分别为1.19%和1.35%,批间精密度CV值分别为1.38%和1.47%,所有靶标精密度CV值均<5%;检测限验证实验中,弱阳性参考品重复测定20次的检出率均为100%;特异性验证实验[对加入严重急性呼吸综合征（SARS）冠状病毒、中东呼吸综合征（MERS）冠状病毒、冠状病毒229E型、冠状病毒OC43型、冠状病毒HKU1型、冠状病毒NL63型、H1N1流感病毒、乙型流感病毒、鼻病毒混合质粒的5份阴性标本检测]中,新型冠状病毒核酸检测结果仍旧均为阴性。结论 该新型冠状病毒核酸检测系统的各...     

鼠类携带汉坦病毒的核酸检测及核酸序列分析

目的 研究国境口岸地区鼠类自然携带汉坦病毒( Hantavirus,HV)状况,为国境口岸地区鼠类防治及肾综合征出血热疫情控制提供基础数据.方法 利用分子生物学技术,通过对采集于口岸地区的鼠类样品的核酸(RNA)提取、逆转录聚合酶链反应(RT-PCR)、凝胶电泳分析以及核酸序列分析等过程,对口岸地区鼠类携带汉坦病毒的状...     

基层医院多重医院感染的病原菌分布及耐药分析

目的 了解基层医院多重医院感染病原菌的临床分布及对常用抗菌药物的耐药情况,以指导临床的合理用药.方法 对2005年1月～2008年10月住院患者中多重医院感染病例分离到的病原菌,进行菌种分布和耐药性的回顾统计分析.结果 临床所分离的284株病原菌中,革兰阴性菌132株,占46.48%,革兰阳性菌84株,占29.58%,真菌48株,占16.90%;前5位病原菌依次为葡萄球菌属、假单胞菌属、真菌、鲍氏不动杆菌和肺炎克雷伯菌;病原菌耐药显示革兰阳性菌除万古霉素、替考西林、夫西地酸、呋喃妥因较敏感外,其余大多耐药率＞60.00%;革兰阴性菌对美罗培南、亚胺培南、阿米卡星、哌拉西林他唑巴坦的耐药率较低外,对其他常用抗菌药物的耐药率大多＞50.00%.结论 基层医院多重医院感染的病原菌仍以革兰阴性菌为主,病原菌耐药严重;重视病原菌检查及药敏监测,对控制多重医院感染及临床合理应用抗菌药物有指导意义.     

郑州市新型冠状病毒核酸检测室间质量评价分析

目的 利用室间质量评价（EQA）评估2020年郑州市实验室新型冠状病毒核酸检测项目的检测能力。方法 对EQA样本采用实时荧光RT-PCR法进行验证,要求参评实验室领取样本2天内完成核酸检测及结果回报,对回报结果按说明书判读条件进行EQA分析。结果 EQA样本可进行实验室间的检测能力评价。31家参评实验室有效回报率为100.00%,29家实验室回报结果完全正确,合格率为93.54%;2家第三方医学检验所成绩为不合格。疾控机构合格率100.00%高于第三方医学检验所89.47%。1个样本检测合格率为93.54%,其他4个样本检测合格率为100.00%。2家不合格检验所整改后成绩合格。结论 郑州市参评实验室新型冠状病毒核酸检测质量整体较好,但个别实验室检测能力尚需提高。     

核酸检测及相关技术在食源性致病菌快速检测中的研究

传统的细菌分离、培养与鉴定由于需时较长,难以适应食源性疾病预防控制的需要。近年来,伴随着核酸检测技术的迅猛发展,各种能够快速检测食源性致病菌的方法相继诞生。本文就聚合酶链式反应及其衍生技术、核酸恒温扩增技术、寡核苷酸微阵列技术、免疫磁性细胞分离技术及DNA生物传感器技术在沙门菌、金黄色葡萄球菌、肠出血性大肠埃希菌等食源性致病菌快速检测中的应用研究进行综述。     

Application of cusum techniques to the routine monitoring of analytical performance in clinical laboratories

Quality control data from a radioimmunoassay of progesterone are analysed. The measurements are found to have a lognormal distribution. A likelihood analysis of the change-point is described. Some rival continuous inspection schemes are compared and it is pointed out that more efficient schemes than Shewhart's can be implemented in clinical laboratories with the aid of a computer.     

急性重度苯酚中毒致心脏骤停1例报告

报道1例急性重度苯酚中毒致心脏骤停,经及时清洗患肢、心肺复苏、连续性静脉-静脉血液透析(CVVHD)联合血液灌流(HP)等对症支持治疗后恢复良好的病例,为该类毒物中毒临床急救提供参考。     

Quality of nuchal transluccency measurements: an exploratory study into their performance and evaluation

OBJECTIVE: To investigate the quality of nuchal translucency measurements that were done as part of a randomised study into screening for Down's syndrome. DESIGN: Exploratory. METHOD: In the period May 2001-April 2003, 396 nuchal translucency measurements were done by 55 sonographers in 13 different echography centres. The sonographers were questioned in writing regarding their training and experience, the procedure and technique used, and the quality assurance. A filled-in questionnaire was received from 42 sono0 images at random from the 2710measurements (70%o) that had been made in 6 centres. These were evaluatedby 3 experts on the basis of the 5 criteria that are used by the Fetal Medicine Foundation (FMF). Each measurement was given 2 points for every criterion that was fulfilled, after which the average of the 3 scores was calculated. RESULTS: Of the 42 respondents, 88% had taken a nuchal translucency measurement course and 45% were certified by the FMF. Calipers were placed incorrectly by 50% of the respondents; 62% of the respondents mentioned the absence of an intern quality check. Concerning the quality-assessment of the nuchal translucency measurements, the assessors unanimously considered i3%o of the 30 images to be suitable for prenatal screen4ngand 43% to be unsuitable; their opinions as to the remaining images differed. The average quality mark was 4.7. CONCLUSION: Many nuchal translucency measurements did not fulfil the quality criteria that are imposed in order to make the measurement reliable as a screening test. There was a lack of quality securance in the echography centres.     

The influence of matrix type, diurnal rhythm and sample collection and processing on the measurement of plasma β-amyloid isoforms using the INNO-BIA plasma Aβ forms multiplex assay

Objective  The aim of the study was to determine the extent to which plasma matrix types, diurnal rhythm and sample collection and processing procedures contribute to overall variability of measurements with the INNO-BIA plasma Aβ forms assay. Methods  Plasma samples from healthy volunteers were collected at BARC-CRI. Analyte concentrations from various plasma matrix types (EDTA, heparin, fluoride) were compared to serum after collection of blood in commercial plastic and glass tubes. Sample processing variables including time and temperature before and after centrifugation, centrifugal force and plasma dilution factor were also investigated. Diurnal variability in plasma Aβ isoforms was determined in 29 healthy volunteers by analysis of EDTA plasma specimens serially collected over 24 hours and stored frozen following oral administration of a placebo treatment. All plasma samples from a given individual and experiment were analyzed in a single analytical run. Results  Highest Aβ levels were obtained using EDTA-plasma samples (in contrast to serum, heparin, citrate, or fluoride). Addition of aprotinin to EDTA plasma had no effect on Aβ peptide recovery. The elapsed time and temperature exposure, before and after sample processing affects the recovery of Aβ isoforms. Analyte recovery was not significantly affected by the presence of platelets in plasma samples. At the subject level, analysis of serially collected EDTA plasma specimens from healthy volunteers revealed no evidence of diurnal variation in any of the Aβ isoforms investigated and results from samples collected on a monthly basis showed only very limited intra-individual variation. Conclusions  Optimal recovery of Aβ peptides was obtained from blood drawn into EDTA tubes and processed within 4 h. Plasma that was refrigerated after separation and analysed within 4 h gave comparable results to samples immediately processed and frozen at −70 °C.     

乙型肝炎病毒的亚克隆及其应用

目的 应用酶切pBP₁₃₂₂HBV,回收HBVDNA,将其克隆到高拷贝载体,获得亚克隆株pUC₁₉HBV,再酶切、HBVDNA标记探针,核酸杂交检测临床血清标本72份,与定量PCR,定性PCR方法对比.方法 由此获得了亚克隆株pUC₁₉HBV,用地高辛标记的HBVDNA探针杂交、定量PCR、定性PCR方法同时检测临床血清标本72份.结果 阳性标本份数分别为40、51、49份,阳性率分别为55.6%、70.8%、68.1%.阳性符合率分别为88.9%、84.6%、80.8%.结论 3种HBV核酸检测方法的比较,以定量PCR法最为敏感,地高辛标记HBVDNA探针检法为特异.     

Genotoxicity evaluation of effluents from textile industries of the region Fez-Boulmane, Morocco: a case study

In order to investigate the biological hazard of effluents from textile industries of Fez-Boulmane region in Morocco, mutagenicity and phytotoxicity tests were performed on different biological systems. Moreover, the efficiency of a Sequencing Batch Reactor (SBR) system, working by activated sludge on a laboratory scale, was estimated by comparing the ecotoxicity results observed before and after wastewater treatment. Evaluation of the genotoxic potential was investigated by means of classic mutagenicity tests on D7 strain of Saccharomyces cerevisiae and by phytotoxicity tests on Allium sativum L., Vicia faba L. and Lactuca sativa L., estimating micronuclei presence, mitotic index and cytogenetic anomalies. The results obtained by testing untreated wastewater demonstrated major genotoxicity effects in S. cerevisiae and various levels of phytotoxicity in the three plant systems, while after SBR treatment no more ecotoxicological consequences were observed. These data confirm the effectiveness of the SBR system in removing toxic substances from textile wastewaters in Fez-Boulmane region.     

Arsenic species excretion in a group of persons in northern Germany--contribution to the evaluation of reference values

Reference values for four arsenic species (inorganic As(III); inorganic As(V); dimethylarsinic acid DMA; monomethylarsonic acid MMA) in urine were evaluated for 101 male persons in northern Germany (46.9 ± 10.5 y) applying anion exchange chromatographic species separation with on-line hydride-technique atomic absorption spectrometry (between-days imprecision 6.8-10.1 %; 11.0-50.0 μg/l; n = 30; detection limits d. l. 1; 10; 2; 2 μg/l). DMA was found in 88.1 % of the persons (x ± s = 7.21 ± 9.64 μg/g creatinine; median 4.02 μg/g; 95 % < 22.5 μg/g) followed by As(III) (11.3 %; median < d. l.; 95 % < 0.54 μg/g) whereas no MMA and As(V) could be found. Seafood consumption within the last 2 days led to higher DMA levels compared to having seafood more than 6 days ago (n = 43 vs. n = 42; 10.04 ± 11.58 vs. 3.47 ± 3.55 μg/g; p < 0.01).     

综合分析方法在妇幼保健机构效率和绩效评估中的应用

目的:应用综合分析方法对广东省地级市妇幼保健机构的运营效率和综合绩效进行评估,对综合分析方法的适用性和绩效评价体系进行验证。方法:采用数据包络分析方法对21所地级市妇幼保健院的投入、产出指标进行分析,评价其效率。采用TOPSIS法与RSR法相结合对21所地级市妇幼保健院进行综合绩效评价和分档。结果:数据包络法分析显示,14所的效率值为1,DEA有效;7所的效率值小于1,DEA无效。TOPSIS法结合RSR法分档,6所机构被评为上档,9所被评为中档,6所被评为下档。结论:数据包络分析方法特别适合多输入、多输出的评价系统,适用于保健机构投入、产出效率的分析。采用TOPSIS法评价结果与实际情况较为符合,评价指标体系较为合理。