临床免疫检测中标准品质量变异的回顾性分析和思考

随着基础医学的发展和高新技术的应用.各类自动化免疫分析仪器相继应用于临床免疫检测.它们基本是原装仪器、配套试剂、专用校准品和设定的操作程序等的组合.形成了一个全封闭的检测系统[1],是保证检测结果准确的一个可靠、有效的基础.这其中,标准品的准确性是保证整个检测系统准确有效的基础,标准品的量值通过定标过程而传递给受检标本.     

临床免疫检测中标准品质量变异的回顾性分析和思考

随着基础医学的发展和高新技术的应用．各类自动化免疫分析仪器相继应用于临床免疫检测．它们基本是原装仪器、配套试剂、专用校准品和设定的操作程序等的组合。形成了一个全封闭的检测系统．是保证检测结果准确的一个可靠、有效的基础。这其中。标准品的准确性是保证整个检测系统准确有效的基础．标准品的量值通过定标过程而传递给受检标本。在临床检验实践中．由于储存运输和使用不当等原因使校准品降解、浓缩或活性、亲和性等方面的变异不是偶然的,是一种经常性现象,只是程度不同而已．正是由于标准品的质量变异而导致量值传递的误差．影响了检测结果的准确性。某些程度严重或不同浓度梯度的标准品间非比例性关系的变化等质量变异因为不能通过定标而被排除．真正影响检测质量的是那些虽发生了质量变异但仍能通过定标的标准品。     

UF-50检尿红细胞结果干扰因素分析

目的探讨草酸钙、细菌酵母在UF-50型尿沉渣全自动检测仪分析中对红细胞检测的干扰。方法采用UF-50型尿沉渣全自动检测仪分析法和镜检法分别分析了2320例住院部患者晨尿。结果2320例中UF-50型沉渣全自动检测仪检出尿红细胞异常264例，镜检法检出217例，与镜检法比较，47例假阳性标本中UF-50检测草酸钙结晶占64％，类酵母菌占14．9％，大量杂菌占12．8％，精子占4．26％，其他药物结晶及杂质占4．26％。结论在UF-50型尿沉渣全自动检测仪分析中，草酸钙、非晶形盐、菌尿、酵母菌对红细胞检测的结果干扰很大，应引起重视。     

一般尿液检测中的干扰因素

尿液检测在泌尿系统疾病及一些全身性疾病的诊治中具有很重要的意义,因而尿液分析是各医院必做的常规项目。随着科技的不断发展,自动化仪器的广泛使用,赋予了尿液分析许多新的内容,但在当前尿液一般检测工作中,有诸多值得检验人员重视并提请医师注意的问题。如尿液检验11项（蛋白、pH、葡萄糖、尿胆原、胆红素、隐血、酮体、亚硝酸盐、密度、维生素C、白细胞）为干化学试纸法,有诸多影响尿液检测的干扰因素。     

氯离子选择电极的干扰因素分析

我们在使用ISE法测定血清钾、钠、氯离子浓度过程中对影响氯电极的干扰因素，进行了如下探讨。     

异嗜性抗体干扰甲状腺激素检测结果的临床病例分析

目的分析和处理异嗜性抗体(HA)干扰甲状腺激素检测结果。方法选取体检健康者血清标本作为对照标本,与病例血清标本同做免疫球蛋白G(IgG)亚型、免疫球蛋白A(IgA)、免疫球蛋白M(IgM)、C-反应蛋白、类风湿因子、抗链球菌溶血素0、转铁蛋白、总蛋白、清蛋白、球蛋白、白球比的测定;采用聚乙二醇(PEG)沉降法对二者标本处理后,再次测定二者的甲状腺激素水平,并计算回收率。结果排除IgG、链霉亲和素、类风湿因子、血清蛋白质等因素对甲状腺激素检测造成的影响;对照标本三碘甲状原氨酸(T3)、甲状腺素(T4)、游离三碘甲状原氨酸(FT3)、游离甲状腺素(FT4)、促甲状腺激素(TSH),PEG沉降回收率均>60%,说明没有大分子蛋白的干扰;病例标本T3、T4、FT3、FT4的PEG沉降回收率均<40%,说明存在大分子蛋白的干扰,TSH的PEG沉降回收率为58.7%,可能存在蛋白的干扰。结论该患者甲状腺激素各指标检测受到HA的干扰,当发现患者临床表现和实验室数据不一致时,在排除其他干扰因素的情况下,可采用PEG沉降法以有效解决此类干扰作用。     

振幅整合脑电图干扰因素分析与护理对策

振幅整合脑电图（aEEG）是一种在病房床旁进行的脑功能监测手段,可为新生儿脑功能状态（包括脑损伤和脑发育情况）的诊断及预后判断提供依据,操作方便、图形直观、容易分析,尤其适合于重症监护病房新生儿脑功能监测。但操作过程中的一些干扰因素会导致图形变异,影响结果判断。因此,排除干扰因素对结果判断的影响非常重要。现将我院新生儿重症监护病房aEEG监测的干扰因素与对策总结如下。     

利用反应曲线识别尿蛋白定量检测中的样品干扰

尿蛋白定量检测对肾脏疾病的诊断治疗有着重要意义。检测尿蛋白定量的方法很多，不论哪种检验方法都有可能受到干扰，受到干扰的检测结果不是对标本客观真实的反映，必然会影响到疾病的诊断治疗。为确保检验结果准确，如何识别和排除干扰是检验工作中的重要内容之一。本文就我科利用反应曲线识别和排除24小时尿蛋白定量检测中的样品干扰进行报道。     

Analytical interferences and analytical quality

In today's health care system the prevalence of medical errors is high as stated by the report of the Institute of Medicine. A varying error rate of < 10% in clinical medical laboratories has been reported in the literature. Most of these errors occur in the pre-analytical phase. Only a small number of errors will be seen in the analytical phase. This overview will deal with the analytical interferences and will offer ways to improve the analytical quality. Some special areas of the analytical process like calibration, quality control, reference interval, drug interference, statistical analysis and volume displacement will be covered. With some examples from the literature and own investigations the impact of errors in these steps of the analytical process will be better understood and the examples will help reducing the number of analytical errors and interferences. This finally provides better patient safety.     

Causal attribution,coping strategies,and learned helplessness

This study assesses the intervening role of coping strategies in mediating the effects of causal attribution for failure, expectancy of control, and off-task cognitions on performance. In study 1, a preliminary investigation, subjects were asked to recall a failure and then to report its major cause and how they coped with it. In study 2, subjects reported their habitual attributional style for negative events and were exposed to failure. Then their expectancy of control, off-task cognitions, coping strategies, and performance were assessed. Results showed that problem-focused coping was associated with less stable/global attribution for failure and with higher expectancy of control. Emotion-focused coping and distancing coping were associated with more internal/global/stable attribution for failure and more frequent off-task cognitions. In addition, distancing coping was found to have a direct effect on performance and to mediate performance effects of causal attribution. The results were discussed in terms of the Lazarus and Folkman stress-coping model.     

20例再生障碍性贫血的细胞因子和粘附分子的表达分析

目的 评价造血微环境中细胞因子对再生障碍性贫血（AA）发病的影响及其临床意义。方法 对20例AA患者及8名正常人用酶联免疫法检测骨髓和外周血中粒细胞集落刺激因子（G－CSF）、肿瘤坏死因子－α（TNF－α）水平；用免疫荧光法检测骨髓和外周血单个核细胞血小板内皮细胞粘附分子（CD31）、淋巴细胞归巢受体（CD44）抗原表达水平。结果 AA组骨髓及外周血G－CSF、TNF－α水平均明显高于正常对照组（P＜0.01)；而CD31、CD44抗原水平却明显低于正常对照组（P＜0.01)。相关分析发现：AA组骨髓及外周血中，G－CSF与外周血白细胞数有髓中粒系比例呈负相关；TNF－α与外周血白细胞数、血小板、血红蛋白及骨髓中粒系比例、红系比例、巨核细胞数均呈负相关；CD31^ 、CD44^ 细胞数与外周血白细胞数、血小板、血红蛋白以及骨髓中粒系比例、红系比例和巨核细胞数呈正相关。结论 造血微环境异常及细胞因子网络失调可能在AA发病中起一定作用。     

Differential interferences of hemoglobin and hemolysis on insulin assay with the Abbott Architect®-Ci8200 immunoassay

Objectives

This paper evaluates the effect of hemoglobin (Hb) and hemolysis on insulin measurements with the Architect®-Ci8200 analyzer.

Design and methods

Insulin concentrations were measured using the Architect®-Ci8200. Interference studies were performed by spiking serum pools of defined insulin concentrations with increasing concentrations of either free Hb or hemolysate. A change of more than 10% was taken as evidence of significant interference.

Results

A significant negative bias in insulin results was observed only in samples spiked with hemolysate but not with free Hb. The bias was proportional to the degree of hemolysis and to the time elapsed before insulin assay. This interference was decreased when samples were kept at + 4 °C.

Conclusions

For all insulin requests, hemolysis must be systematically checked before biological interpretation of insulin results.     

Clinically insignificant negative interferences of spironolactone, potassium canrenoate, and their common metabolite canrenone in new dimension vista Loci digoxin immunoassay

Spironolactone, a potassium-sparing diuretic metabolized to canrenone is often used with digoxin to treat various conditions including congestive heart failure. Potassium canrenoate is a similar drug, which is also metabolized to canrenone. Due to reported both positive and negative interference of spironolactone, potassium canrenoate, and their common metabolite canrenone with digoxin immunoassays, we investigated potential interference of these compounds with the new homogenous sequential chemiluminescent assay for digoxin based on the luminescent oxygen channeling technology (LOCI digoxin) for application on the Dimension and Vista platform. When aliquots of a drug-free serum pool were supplemented with various amounts of spironolactone, potassium canrenoate, or canrenone and apparent digoxin values were measured using Dimension Vista LOCI digoxin assay, we observed no detected value except when aliquots were supplemented with very high amounts of potassium canrenoate or canrenone. However, we observed that apparent digoxin concentrations were very low. When aliquots of a serum digoxin pool (prepared by pooling specimens from patients receiving digoxin), were further supplemented with various amounts of spironolactone, potassium canrenoate, or canrenone and serum digoxin concentrations were remeasured using the LOCIdigoxin assay, only statistically significant falsely lower digoxin values (negative interference) were observed in specimens containing very high amounts of canrenone or potassium canrenoate. However, such small bias may not have any clinical significance. We conclude that new Dimension Vista LOCI digoxin assay is virtually free from interferences of spironolactone, potassium canrenoate, and their common metabolite canrenone.     

survivin基因RNA干涉对宫颈癌裸鼠移植瘤生长及凋亡和顺铂化疗敏感性的影响

目的观察survivin基因RNAi对宫颈癌裸鼠移植瘤生长、凋亡和化疗敏感性的影响。方法随机选择雌性BALB/C-nu/nu裸小鼠24只,细胞接种法建立4组人宫颈癌裸鼠皮下移植瘤模型,每天观察裸鼠一般状况及肿瘤生长情况,通过绘制肿瘤生长曲线并计算肿瘤生长抑制率,观察survivin基因RNAi对人宫颈癌裸鼠皮下移植瘤生长的影响;通过免疫组化SP法检测各组移植瘤组织中survivin蛋白表达情况,TUNEL染色观察survivin基因RNAi对人宫颈癌裸鼠皮下移植瘤凋亡的影响;当肿瘤体积达0.2cm3时给予顺铂化疗以观察survivin基因RNAi对人宫颈癌裸鼠皮下移植瘤化疗敏感性的影响。结果成功建立4组人宫颈癌裸鼠皮下移植瘤模型,接种HeLa-s2组裸鼠肿瘤体积在每个检测点均明显小于接种HeLa组;观察结束时,接种HeLa-s2组裸鼠瘤重明显小于接种HeLa组,分别为：（0.369±0.043）g和（1.150±0.136）g（P〈0.05）;接种HeLa-s2组裸鼠肿瘤生长抑制率为67.9%。免疫组化结果显示：接种HeLa-s2组裸鼠survivin蛋白表达显著下降;TUNEL染色结果显示：接种HeLa-s2组裸鼠细胞凋亡明显增多,凋亡指数（AI）值达（22.73±1.37）%。顺铂化疗后不同检测点接种HeLa-s2组裸鼠肿瘤体积明显小于接种HeLa组,肿瘤生长明显受抑;观察结束后,接种HeLa-s2组裸鼠瘤重明显小于接种HeLa组,分别为：（0.323±0.058）g和（1.347±0.173）g（P〈0.05）;接种HeLa-s2组裸鼠肿瘤细胞凋亡明显增多,与接种HeLa组AI比较,接种HeLa-S2组AI明显升高,分别为：（37.38±1.01）%和（5.19±0.61）%（P〈0.05）。结论 survivin基因RNAi可通过下调移植瘤组织survivin蛋白表达抑制移植瘤生长并促进其凋亡,并通过增加顺铂化疗诱导的细胞凋亡,增强顺铂化疗对移植瘤的生长抑制,进而提高移植瘤对顺铂化疗的敏感性。     

免疫透射比浊法常见干扰因素的识别与应对策略

免疫透射比浊法检测结果的准确性常受到一些干扰因素的影响,如样本性状、内源性干扰、钩状效应和携带污染等,这些干扰因素可使检测结果假性升高或降低,影响临床诊断和治疗监测等.文章对免疫透射比浊法常见的干扰因素及机制进行综述,以帮助检验人员识别和确认可能存在的干扰,及时纠正检测结果,尽可能减少不良后果的发生.     

免疫监测研究与应用中值得关注的几个问题

免疫监测是免疫检验中的重要组成部分,包含从细胞到生物分子等多种指标.在此重点讨论3个问题:首先,T淋巴细胞是所有免疫应答的中心环节,对其进行检测,对了解免疫相关疾病非常重要.在临床检验中应根据实验室条件(如流式细胞仪)和检测目的 (如抗原特异性T淋巴细胞数量或功能)选择不同方法.其次,自身抗体是诊断自身免疫性疾病的重要指标;有些自身抗体甚至可预测疾病的发生,越来越多研究表明,肿瘤患者中存在多种自身抗体,这些自身抗体有可能对肿瘤诊断、治疗肿瘤靶分子的筛选提供帮助.另外,检测细胞因子是了解疾病免疫学机制的重要途径;细胞因子检测有助于判断疾病活动程度,但不具有器官或组织特异性;应加强细胞因子检测的质量控制,以提高其在临床检验中的应用质量.     

LC-MS/MS quantitation of ribavirin in serum and identification of endogenous isobaric interferences

BackgroundRibavirin is a nucleoside analog used in treatment of chronic hepatitis C. It is associated with severe, dose-dependent toxicities, including hemolytic anemia. To facilitate therapeutic drug monitoring, a liquid chromatography–tandem mass spectrometry method was validated for quantitation of ribavirin in serum.MethodsAfter protein precipitation, ribavirin is quantitated using a ¹³C₅-ribavirin internal standard, on a Hypercarb analytical column designed for retention of polar analytes.ResultsThe analytical method shows excellent precision, sensitivity, and specificity. In vitro drug stability was also assessed. Interestingly, endogenous isobaric compounds were noted in both human and bovine serum; these could be chromatographically separated from the ribavirin peak. Addition of exogenous uridine and cytosine increases the size of the isobaric peaks, suggesting that these compounds are the source of the endogenous interference.ConclusionsThis method uses mass spectrometric transitions that have been used in other published methods, but also separates ribavirin from isobaric peaks that were not described. These peaks were determined to be endogenous nucleosides. Laboratories quantitating ribavirin in biological matrices should be aware of the potential for isobaric interferences, and take steps to chromatographically separate them from the ribavirin peak for accurate quantitation.     

1例罕见B（A）血型鉴定及其临床安全输血策略初探

目的 鉴定罕见B（A）血型并探讨其临床安全输血策略。方法 利用全自动血型分析仪检测ABO血型,发现正反定型不符的一位患者,利用血型血清学和分子生物学方法对该患者及其家系中9位成员进行血型鉴定和ABO血型基因测序,分析该家族血型的遗传特点;并对该血型的患者进行临床输血探讨,确保临床输血安全。结果 患者为B（A）血型,与B101基因序列比对,均为nt640A>G,基因型鉴定为B(A)04/O01,B(A)型血清与同型红细胞以及O型、B型红细胞不凝集,B(A)型红细胞与同型以及AB型血清不凝集。结论 血型血清学方法可以对正反定型不符的B（A）血型做出初步判断,分子生物学技术可以进一步准确判断该血型;临床输血应采取自体输血、同型输注及配合相容性输血。