ELISA法检测HBsAg室内质控方法及结果分析

目的 探讨ELISA法测定HBsAg的室内质控方法 .方法 选择一个阳性(0.5mg/ml)质控品和一个阴性质控品,采用L-J的室内质控方法 和对阳性、阴性质控品进行检测,采用12S、13S质控规则和Icutoff(+)、Icutoff(-)质控规则对ELlSA法测定HbsAg进行室内质控.结果 检测0.5ng/ml质控血清20天,得20个s/co值,经统计得20个s/co值的X为1.984,SD为0.551,CV为27.8%.2月份74个阳性(0.5ng/ml)质控结果中,最大的s/co值为3.01,最小值为0.952,按12S、13S质控规则无失控和警告结果,而按Icutoff(+)有一次失控,2月份74个阴性质控品测定结果中最大值为0.653,均小于1,按Icutoff(-)质控规则本月阴性质控品无失控,即无假阳性结果 .3月份96个s/co结果 的时段均值(x)为1.746,时段标准差(SD)为0.5385,时段CV为30.8%,按12S、13S质控规则仅有一次警告,按Icutoff(+)规则也无失控.3月份96个阴性质控品测定结果 中有一次阴性质控品检测的s/co结果 大于1.按Icutoff(-)规则判断本批次为失控.结论 ELISA检测试剂盒生产厂商应标明试剂的灵敏度或检测低限,ELISA法的室内质控品应选择一个灵敏度(或检测低限)浓度处的室内质控品,采用于Icutoff(+)规则用于监控检测结果 中的假阴性;同时选择一个阴性质控品,采用Icutoff(-)用于监控检测结果 的假阳性,不能按统计学方法 来选择什么样浓度的质控品或按经验选择s/co值在2～4相当浓度的质控品,而用L-J室内质控方法 进行ELISA法检测过程的室内质控,本作者认为意义不大,但其可用于评估和监测ELISA法检验过程中的精密度.     

自制人类免疫缺陷病毒抗体等质控血清及室内质控方法建立研究

目的用人类免疫缺陷病毒(HIV)抗体阴性血清梯度稀释HIV抗体酶联免疫试剂盒中阳性对照,制备室内质控血清。建立艾滋病酶联免疫检测的室内质控方法。进行方法可靠性和质控血清稳定性的评价,同时比较两个厂家生产的HIV抗体试剂盒进行自制质控血清室内质控结果以及自制丙型肝炎抗体、乙型肝炎表面抗原血清的室内质控结果的评价。方法将自制的HIV质控品与卫生部HIV质控品按照HIV抗体检测的SOP文件用HIV抗体试剂随患者进行检测,每天测定吸光度,计算S/CO値。连续检测20次计算S/CO值和s,以x±2s为控制限,以x±3s为失控限绘制质量控制框架图。每天随患者标本各加入1份自制质控品和1份卫生部购质控品进行质控将其各自的S/CO描在质控图上,应用多规则质量控制原则分析以发现当天测定批的误差以及误差原因。控制每批次检测的重复性,观察试剂盒间的差异进行两种质控血清质控结果的比对研究。结果自制HIV抗体质控血清方法科学可靠,减少了基质效应,能确保结果的准确性和可靠性。结论稀释阳性对照制备质控品,建立酶联免疫室内质控方法也适用于乙型肝炎表面抗原、丙型肝炎抗体等其他免疫项目的酶联免疫室内质控。不同厂家试剂用自制质控品进行质控结果无差异。     

荧光定量PCR测定HBV DNA室内质控方法的探讨

目的探讨实时荧光定量聚合酶链反应(PCR)测定乙型肝炎病毒核酸(HBV DNA)的室内质控方法。方法统计上海地区PCR实验室HBV DNA常规条件下前20次室内质控数据的不精密度(CV),以测出的均值(x珋±s)绘制质控图,分别采用13s/22s的多规则质控方法和Levey-Jennings单规则质控方法,判断前20次室内质控数据CV分别为≥10%、5%~10%和1%~5%范围内A、B和C 3家实验室的室内质控数据,分别分析其失控检出能力。结果分别采用13s/22s多规则和Levey-Jennings单规则质控方法。当HBV DNA室内质控品低、高两个浓度分别为5×104和5×106IU/mL时,CV为14.96%和12.15%的A实验室均未正确检出失控数据;CV为6.49%和5.00%的B实验室检出2个随机误差引起的失控数据;其CV为4.36%和2.43的C实验室,采用13s/22s多规则质控方法检出3个系统误差引起的失控数据,采用单规则质控方法未检出失控。结论 PCR检测HBV DNA当实验室前20次室内质控数据CV≥10%时,无论采用单规则还是多规则质控方法均不能正确检出失控;实验室应设定自己实验室最低要求的CV,并采用多规则质控方法,以提高系统误差的失控检出率。     

定量检测BCR-ABL(P210)转录本水平室内质控体系的完善及多中心推广应用北大核心CSCD

目的建立完善的BCR-ABL(P210)监测室内质控体系, 确保检测结果的长期稳定性和室间可比性, 并在3家医院进行推广应用。方法山东大学齐鲁医院血液病研究室(H1)利用实时定量PCR(RQ-PCR)检测质控物的BCR-ABL(P210)转录本水平, 完善质控体系的可靠性和稳定性;赴其他3家医院(H2~H4)现场进行测定前质量控制检查, 同时邮寄给各家医院25套质控物进行检测;采用Levey-Jennings质控图结合Westgard多规则方法, 对各家医院的标准曲线斜率、截距及检测结果进行统计学的质量判断, 作出质控评价。结果①成功建立并完善了测定前质控检查、测定中质控统计判断、测定后质控评价的BCR-ABL(P210)转录本水平监测的室内质控体系。②4家医院标准曲线的斜率、截距均在控。③多中心质控物判断结果:H1医院中、低浓度质控物各出现1次"12s"警告, 判断为在控;H2医院3种浓度质控物各出现1次"12s"警告, 判断为"22s"失控;H3医院高浓度质控物违反1次"13s"规则, 低浓度质控物出现1次"12s"警告, 判断为"13s"失控;H4医院各质控物均在控。④质控评价及纠正:2家医院在控, 另2家医院各出现1次失控, 查找原因进行纠正后未再出现失控。⑤多中心质控物原始数值比较:4家医院之间高浓度质控物结果差异有统计学意义, 中、低浓度质控物结果差异无统计学意义。⑥多中心质控物国际标准值(IS值)比较:H2、H3医院的IS数值显著高于H1医院, H2医院显著高于H3医院。结论建立了一套完善稳定的BCR-ABL(P210)转录本室内质控体系, 有效地保证临床检测结果的稳定性和室间可比性, 并成功完成了多中心的推广应用。     

解脲支原体DNA检测室内质控物的制备及质控方案的设计

目的制备解脲支原体(UU)DNA荧光定量PCR室内质控物,建立室内质量控制体系,对其临床应用进行初步评价。方法分别留取Ct值为24~25(阳性)和32~33(弱阳性)标本和检测结果为阴性的标本,待留取到15mL时充分混匀,按每管150μL分装作为室内质控物。前20次检测采用"即刻法"判断检测结果是否在质控范围内,20次以后绘制Levey-Jennings图,确定靶值、标准差(s)和变异系数(CV),采用Westdard多规则质控方法对自制室内质控物检测结果进行判断。在Unity Real Time(URT)系统中使用质控规则配置操作导出UU-DNA的功效函数图(OPSPecs图),根据OPSPecs图设置质控规则。结果131次实验中,前20次采用"即刻法"判断室内质控物;20次以后采用Levey-Jennings图判断,质控品稳定,质控规则合理。结论用临床分泌物混合液制备UU-DNA检测项目的室内质控物品,制备方法简单,测定结果稳定,可作为实验室检测UU-DNA项目的质控品;依据OPSPecs图设置分子生物学检测项目的室内质控规则简便、实用。     

无锡地区实验室2010年室内质控情况回顾性分析

目的了解无锡市中心血站实验室室内质控效率及实验室整体检测水平的发展趋势。方法对2010年无锡市中心血站实验室血液检测ELISA试验的室内质控数据进行回顾性分析。结果 2010年本站实验室ELISA检测项目共有5 793个室内质控结果,其中65次(1.12%)属于"警告",20次(0.35%)属于"失控";失控中随机误差7次(35%),系统误差13次(65%)。随机误差和系统误差分别占质控总数的比例为0.12%和0.22%。结论从总体上说明本站实验室室内质控效率处于室内质控的可接受范围内。     

CMV-DNA荧光定量PCR检测室内质控物的制备及初步应用

目的制备巨细胞病毒DNA(CMV-DNA)荧光定量PCR检测的室内质控物,并对其临床应用进行初步评价。方法收集一批CMV-DNA阳性尿液标本混合后作为室内质控物,前20次检测采用"即刻法"判断结果是否在质控范围内,20次后采用Levey-Jennings质控图,确定其靶值、标准差和变异系数,对室内质控结果进行判断。结果 107次试验中,前20次根据"即刻法",室内质控结果在控;20次后采用Levey-Jennings质控图,结果在控;60次检测结果的均值为5.41,标准差为0.33,变异系数为6.15%。结论利用混合尿液标本制备CMV-DNA室内质控物,制备方便,测定结果稳定,可以用作本实验室检测CMV-DNA的室内质控物。     

血站室内质控即刻法在ELISA检测中的应用

目前，血站对献血者进行乙型肝炎表面抗原(HBsAg)，丙型肝炎病毒抗体(抗-HCV)及艾滋病病毒抗体(抗-HIV)的筛选，主要采用酶联免疫吸附试验(ELISA)。要保证检测结果的可靠性，保障供血质景，就必须对ELISA检测开展室内质控。我站应用即刻法对ELISA检测进行室内质控，并绘制了……     

化学发光免疫技术对乙型肝炎两对半定量检测的临床意义

目的应用化学发光免疫技术对乙型肝炎（下称乙肝）两对半定量检测的意义进行探讨。方法应用Chemflex化学发光免疫技术定量检测乙肝两对半。结果补充血清乙肝病毒DNA（HBV-DNA）荧光定量聚合酶链反应测定可以直接反映病毒复制状态,具有临床应用价值,但有不能完全反映病毒复制静息肝细胞内HBV病毒状态的缺点。结论乙肝病毒表面抗体的定量测定能够对抗体是否真正具有“中和”HBV的免疫力作出正确评价,对乙肝的预防起到监督作用。     

血液病毒核酸筛查系统室内质控方法的建立及评价

目的建立并评价血液病毒核酸检测系统的室内质控方法。方法将浓度为200 IU/m L的HBV DNA、2 000 IU/m L的HCV RNA及2 000 IU/m L的HIV RNA质控标准品用已知阴性献血者血浆标本作不同倍数稀释,直至接近检测下限,确定合适质控品浓度后与献血者标本一同进行核酸提取、扩增,连续检测20次,用即刻法分析所得核酸扩增循环阈值(Ct值)结果,计算Ct值的均值(x珋)、标准差(s)和变异常数(CV),以x珋±2s为警告限,超过x珋±3s为失控限,以此分别建立核酸筛查室内质控图框架并分析室内质控结果。绘制Levey-Jennings质控图,统计180次室内质控结果,结合Westgard多规则质控方法,评价该室内质控模式的可行性和有效性。结果以HBV DNA为例,室内质控品浓度与Ct值的相关系数r=-0.920,在0.05水平达到高度相关。选择浓度100 IU/m L的HBV DNA室内质控品,连续测定20次的Ct值x珋为31.76,s为1.10,CV为3.46%。连续检测180次室内质控品Ct值的x珋为32.02,s为1.13,CV为3.53%。结论 Ct值可以作为室内质控的依据。本实验室选择的弱阳性质控品能增强当次实验结果的可靠性,可以排除因人员更换等实验条件的差异而导致的误差,可用于日常监控核酸提取扩增检测的有效性。     

Determination of creatine kinase isoenzyme MB activity in serum using immunological inhibition of creatine kinase M subunit activity. Activity kinetics and diagnostic significance in myocardial infarction.

This is a new method for the determination of creatine kinase isoenzyme MB activity in serum. The method uses direct activity measurement of creatine kinase B subunit activity after blocking of CK-M subunit activity by inhibiting antibodies. The test takes no longer than 15 min. The method yields an intra-serial C.V. of 2.0-12.9%, and a C.V. from day to day of 5.5%. The detection limit is 3.4 U/l creatine kinase MB. In the 95 cases with proven myocardial infarction several types of creatine kinase MB activity kinetics could be determined. The percentage of creatine kinase MB of peak CK-total is 6-25%, with a mean of 11.1%. The amount of creatine kinase MB with respect to total CK activity after reinfarction is higher than the amount after initial infarction.     

Dissociable correlates of two classes of retrieval processing in prefrontal cortex

Although substantial evidence suggests that the prefrontal cortex (PFC) implements processes that are critical for accurate episodic memory judgments, the specific roles of different PFC subregions remain unclear. Here, we used event-related functional magnetic resonance imaging to distinguish between prefrontal activity related to operations that (1) influence processing of retrieval cues based on current task demands, or (2) are involved in monitoring the outputs of retrieval. Fourteen participants studied auditory words spoken by a male or female speaker and completed memory tests in which the stimuli were unstudied foil words and studied words spoken by either the same speaker at study, or the alternate speaker. On "general" test trials, participants were to determine whether each word was studied, regardless of the voice of the speaker, whereas on "specific" test trials, participants were to additionally distinguish between studied words that were spoken in the same voice or a different voice at study. Thus, on specific test trials, participants were explicitly required to attend to voice information in order to evaluate each test item. Anterior (right BA 10), dorsolateral prefrontal (right BA 46), and inferior frontal (bilateral BA 47/12) regions were more active during specific than during general trials. Activation in anterior and dorsolateral PFC was enhanced during specific test trials even in response to unstudied items, suggesting that activation in these regions was related to the differential processing of retrieval cues in the two tasks. In contrast, differences between specific and general test trials in inferior frontal regions (bilateral BA 47/12) were seen only for studied items, suggesting a role for these regions in post-retrieval monitoring processes. Results from this study are consistent with the idea that different PFC subregions implement distinct, but complementary processes that collectively support accurate episodic memory judgments.     

俯卧位通气对高海拔地区肺复张治疗无效急性呼吸窘迫综合征患者氧合的影响

目的 探讨俯卧位通气对高海拔地区肺复张术(RM)治疗无效急性呼吸窘迫综合征(ARDS)患者的治疗作用.方法 从海拔2260m的地区医院筛选RM治疗无效的41例ARDS患者[平均氧合指数( PaO2/FiO2)较RM前升高＜20％视为RM无效],依不同病因分为肺内源性ARDS组(ARDSp组)和肺外源性ARDS组(ARDSexp组),每组再按信封法随机分为俯卧位组和仰卧位组,即ARDSp俯卧位组(11例)、ARDSp仰卧位组(9例)、ARDSexp俯卧位组(10例)、ARDSexp仰卧位组(11例).在通气前及通气1、2、3、4h监测动脉血氧分压( PaO2)、PaO2/FiO2、静态顺应性(Cst)、气道阻力(Raw)的变化.结果 通气lh时,ARDSexp俯卧位组PaO2/FiO2( mm Hg,l mm Hg=0.133 kPa)即较通气前显著升高(157.4±40.6比129.3±48.7,P＜0.05),并随通气时间延长呈持续增高趋势,4h达峰值(219.1 ±41.1);且ARDSexp俯卧位组通气3h内PaO2/FiO2较其他3组显著增高,另3组间则差异无统计学意义.ARDSp俯卧位组、ARDSexp俯卧位组通气4h时PaO2/FiO2均较相应仰卧位组显著增高(208.8±39.7比127.4±47.1,219.1±41.1比124.9±50.8,均P＜0.05).4组通气前后Cst无显著改变,各组间差异也无统计学意义.ARDSp俯卧位组通气4h时Raw(cmH2O·L-1·s-1)较通气前显著降低(6.8±1.7比10.7±1.8,P＜0.05),且明显低于其他3组;其他3组各时间点Raw组内及组间比较差异均无统计学意义.结论 俯卧位通气作为ARDS机械通气重要策略之一,可以改善RM无效高原ARDS患者的氧合,为抢救患者赢得宝贵的时间.     

Digital imaging among medical facilities

The Department of Veterans Affairs (VA) in the USA operates a network of 172 medical centres which all utilize a hospital information system (HIS) which has been developed and is currently maintained by the VA. During the past several years, an image management and communication module has been developed, installed and clinically utilized at the Washington DC and Maryland VA Medical Centres. This image management and communication system, referred to as the decentralized hospital computer program (DHCP) imaging system, is fully integrated with a commercial picture archiving and communication system (PACS). The system is utilized to capture, archive, and display all images generated within the hospital including radiology, nuclear medicine, pathology, endoscopy, bronchoscopy, and dermatology, intraoperative photographs, ECG data, and a limited number of paper documents. The ultimate goal of the project is to have all patient text and image data available at any clinical workstation to any authorized user anywhere within the network of medical centres. Clinical requirements for an imaging workstation include ease of use, rapid and reliable access to the complete set of patient information, and images which are of acceptable quality to meet the requirements of the user and the subspecialty. Patient confidentiality and data security must be safeguarded at all times. Integration of the images with the remainder of the patient's database was found to be critical to the success of the project. The experience at the Washington and Maryland facilities suggests that an imaging system that is successfully integrated with a hospital information system can provide substantial clinical and economic benefits both within and among medical centres. Clinical acceptance and utilization of the system has been excellent, particularly in diagnostic radiology where DHCP Imaging has been interfaced to a commercial PAC system. Based upon this initial experience, the VA has begun to deploy the system throughout its large network of medical centres.     

Myocardial elastography at both high temporal and spatial resolution for the detection of infarcts

Myocardial elastography is a novel method for noninvasively assessing regional myocardial function, with the advantages of high spatial and temporal resolution and high signal-to-noise ratio (SNR). In this paper, in-vivo experiments were performed in anesthetized normal and infarcted mice (one day after left anterior descending coronary artery [LAD] ligation) using a high-resolution (30 MHz) ultrasound system (Vevo 770, VisualSonics Inc., Toronto, ON, Canada). Radiofrequency (RF) signals of the left ventricle (LV) in longitudinal (long-axis) view and the associated electrocardiogram (ECG) were simultaneously acquired. Using a retrospective ECG gating technique, 2-D full field-of-view RF frames were acquired at an extremely high frame rate (8 kHz) that resulted in high-quality incremental displacement and strain estimation of the myocardium. The incremental results were further accumulated to obtain the cumulative displacements and strains. Two-dimensional and M-mode displacement images and strain images (elastograms), as well as displacement and strain profiles as a function of time, were compared between normal and infarcted mice. Incremental results clearly depicted cardiac events including LV contraction, LV relaxation and isovolumetric phases in both normal and infarcted mice, and also evidently indicated reduced motion and deformation in the infarcted myocardium. The elastograms indicated that the infarcted regions underwent thinning during systole rather than thickening, as in the normal case. The cumulative elastograms were found to have higher elastographic SNR (SNR(e)) than the incremental elastograms (e.g., 10.6 vs. 4.7 in a normal myocardium, and 6.0 vs. 2.4 in an infarcted myocardium). Finally, preliminary statistical results from nine normal (m = 9) and seven infarcted (n = 7) mice indicated the capability of the cumulative strain in differentiating infracted from normal myocardia. In conclusion, myocardial elastography could provide regional strain information at simultaneously high temporal (>/=0.125 ms) and spatial ( approximately 55 microm) resolution as well as high precision ( approximately 0.05 microm displacement). This technique was thus capable of accurately characterizing normal myocardial function throughout an entire cardiac cycle, at the same high resolution, and detecting and localizing myocardial infarction in vivo.     

Clinical Pharmacokinetics of Morphine

Morphine, the most widely used mu-opioid analgesic for acute and chronic pain, is the standard against which new analgesics are measured. A thorough understanding of the pharmacokinetics of morphine is required in order to safely and effectively use this analgesic in a wide variety of patients with different levels of organ function. A MEDLINE search was conducted to identify literature published between 1966 and January 2002 relevant to the pharmacokinetics of morphine. These publications were reviewed and the literature summarized regarding unique and clinically important elements of morphine disposition relative to its parenteral administration (including intravenous, intramuscular, subcutaneous, epidural and intrathecal administration), absorption profile (immediate release, controlled release, and sublingual/buccal, and rectal administration), distribution, and its metabolism/ excretion. Special populations, including infants, elderly, and those with renal/liver failure, have a unique morphine pharmacokinetic profile that must be taken into account in order to maximize analgesic efficacy and reduce the risk of adverse events.