2008至2009年度上海市糖化血红蛋白项目室间质评结果分析

目的分析上海市2008~2009年度糖化血红蛋白（HbA1c）项目室间质评结果,为提高HbA1c检测质量提供依据。方法收集2008~2009年参加上海市临床检验中心HbA1c室间质评医院的4次调查结果,对结果进行统计分析。结果 2008~2009年上海市开展糖化血红蛋白室间质评的医院数从2008年122家增加至2009年178家,合格率从2008年96.7%增加到2009年98.9%。部分的调查结果离散度〉10%[2008年第1次的低压液相（LPLC）1号样本（10.8%）、免疫比浊法2份人新鲜全血样本（15.4%和15.9%）;2008年第2次的免疫比浊法2号样本（10.5%）;2009年第1次的微粒色谱法2号样本（10.3%）;2009年第2次的微粒色谱法2号样本（11.0%）和免疫比浊法1号样本（10.0%）]。经过质控管理,至2009年第2次调查,整体的平均变异系数（CV）有明显改进,人新鲜全血样本各检测系统的结果无明显差异,除微粒色谱法外,符合率达到100%。结论通过加强质控管理,开展HbA1c标准化工作,推动HbA1c检测结果的准确、可比,为临床提供可靠的诊疗依据。     

血红蛋白A2和F的室间质量评价结果分析

目的 评价中国部分珠蛋白生成障碍性贫血筛查实验室检测血红蛋白A2和F的水平和现状。方法 向50家珠蛋白生成障碍性贫血实验室发放两个批号的质控品,收集实验室回报的HbA2和HbF检测值,对回报的结果按照方法分组进行统计分析,并评价其检测水平。结果 49家实验室回报了检测结果,回报率为98%。HbA2的及格率为42.9%～92.3%,HbF的及格率为27.3%～84.6%。通过稳健Z比分数统计,血红蛋白HbA2项目201311批号样本有3家实验室检测结果为不满意,201312批号样本有4家实验室检测结果为不满意,血红蛋白HbF项目201311批号样本有5家实验室检测结果为不满意,201312批号样本有3家实验室检测结果为不满意。结论 我国珠蛋白生成障碍性贫血筛查实验室检测HbA2和HbF质量有待进一步提高。     

HbF变异体对三种不同HbA1c检测系统的干扰评价

目的 评价血红蛋白变异体F(HbF)对三种糖化血红蛋白(HbA1c)检测系统测定结果的干扰。三种方法分别为离子交换高效液相色谱法(IE-HPLC)、硼酸盐亲和层析高效液相色谱法(AC-HPLC)和免疫抑制比浊法。方法 分别用三种检测系统检测血红蛋白结构正常标本及含有胎儿血红蛋白(HbF)的标本,依据美国国家糖化血红蛋白标准化计划(NGSP)的判定标准,对检测结果进行比对分析和偏倚评估。结果 以Primus Ultra2(AC-HPLC)为比较系统,IE-HPLC和免疫比浊法为检测系统,两种检测系统与比较系统检测正常样品HbA1c值的相关性良好。当HbF≤8.75%时,IE-HPLC测定结果与比较系统偏差<6%; HbF为17.50%时,IE-HPLC测定结果与比较系统偏差>6%; 当HbF浓度达35.00%～70.00%时IE-HPLC无法检测出结果。免疫比浊法测定不同HbF浓度HbA1c值与比较系统偏差均<6%,测试几乎不受HbF的干扰。结论 HbF对不同HbA1c检测系统的干扰程度不同,临床实验室在进行HbA1c检测时,应注意血红蛋白变异体的存在,必要时选用替代指标或者合适的方法进行HbA1c测定以防止干扰的发生。     

变异血红蛋白和氨基甲酰血红蛋白对糖化血红蛋白测定干扰的评价

目的评价变异血红蛋白和血红蛋白衍生物氨基甲酰血红蛋白对离子交换-高效液相色谱(HPLC)系统和用酶法检测糖化血红蛋白(HbAlc)的影响。方法分别用2种方法同时检测无尿毒症糖尿病患者血红蛋白结构正常的血液样本和含有不同浓度胎儿血红蛋白(HbF)的血液样本及尿毒症患者HbA1c浓度。结果对于血红蛋白结构正常且无尿毒症的糖尿病患者,2种方法的HbA1c检测结果差异无统计学意义(P>0.05)。当HbF<8.75%时,离子交换-HPLC的HbA1c测定结果与预期值无差异;当样本中HbF浓度为8.8%～23.0%时,其HbA1c测定结果要明显低于理论浓度;当HbF浓度达35.0%～70.0%时,离子交换-HPLC无法检测出结果。因尿毒症患者血液中含氨基甲酰血红蛋白,所以离子交换-HPLC的HbA1c检测结果明显高于酶法;而酶法检测HbA1c不受血液中HbF和氨基甲酰血红蛋白的干扰。结论采用离子交换-HPLC检测含HbF和氨基甲酰血红蛋白的样本中的HbA1c时,结果可能受到干扰;而酶法检测HbA1c时几乎不受样本中HbF和氨基甲酰血红蛋白的干扰。     

2014～2017年我国临床实验室HbA2和HbF检测室内质控变异系数分析

目的 了解2014～2017年我国临床实验室血红蛋白A₂(HbA₂)和血红蛋白F(HbF)检测室内质控情况。方法 收集2014～2017年参加卫生部临床检验中心室间质评参评单位回报的室内质控信息,将变异系数与1/3TEa(6.67%)和1/4TEa(5%)标准进行比较,得到满足各标准实验室的比例,进而分析我国HbA₂和HbF检测的室内质控情况。按照实验室使用的仪器不同进行分组,分别计算2017年HbA₂和HbF不同仪器组在两种标准下的变异系数通过率。结果 HbA₂项目84%以上的实验室精密度都能达到1/3TEa的标准,达到1/4TEa标准的实验室比例略有降低(70.83%～84.47%)。HbF项目的结果除2015年外,当月和累积在控变异系数80%以上的实验室能达到1/3TEa标准,达到1/4TEa标准的实验室比例分布在68.42%～85.07%。2017年数据按仪器分组统计中,两个项目sebia capillarys 2仪器组和全自动血红蛋白分析仪伯乐VariantⅡ仪器组在两种标准下的变异系数通过率均达85%以上,精密度较好。结论 目前我国实验室HbA₂和HbF检测的不精密度水平需进一步提高,尤其是HbF,实验室应继续加强室内质量控制,建立严格的室内质控制度,提高检测水平。     

全自动多通道毛细管区带电泳技术在血红蛋白分析中的临床应用

目的探讨全自动多通道毛细管区带电泳技术在血红蛋白分析中的临床应用价值。方法采用法国SebiaCapillarys2电泳系统（Version5．50）及配套试剂,全血样品经系统自动处理后,在9．8kV电压、34℃、pH9．4的缓冲液条件下,在8条并联的毛细管（17．5cm&#215;25μm）内进行血红蛋白电泳,415nm下直接检测各组分的百分比浓度。结果分析183例健康成人血红蛋白电泳结果,确定本实验室参考范围HbA2：2．1％～3．2％。应用HbAFSC质控进行精密度试验,各组分的cV值在（批内：0．21％～1．07％;批间：0．60％43．10％）之间。检测6045例临床需要进行溶血性黄疸诊断和鉴别诊断的患者标本,检出HbA2增高379例;HbF增高203例;HbH带27例;HbE带8例;HbS带4例;HbJ—K带3例;HbBart’s2例;HbD1例。结论SebiaCapillarys2电泳系统能够清晰分辨HbA2、HbF增高及其他异常区带;可准确地将HbC和HbE与HbA2区分;还可完美区分和聚焦HbA与HbS之间的HbF;并可以轻易区分移动速度相近的HbS和HbD2具有自动化程度高、操作简便、快速准确的特点,适合医疗机构临床实验室在血红蛋白分析中常规应用。     

全国糖化血红蛋白A1c室内不精密度调查

目的调查糖化血红蛋白(HbA1c)室内质控不精密度现状,了解实验室满足允许不精密度质量规范水平的情况,并提出改进措施。方法用基于网络的电子化室间质评上报系统收集2014年参加全国HbA1c室间质评的1 250家实验室的室内质控结果,包括2014年2月和累积的室内质控在控数据的变异系数(CV)等相关信息;用《糖化血红蛋白实验室检测指南》中对于HbA1c精密度的要求[室内CV3%,以2%为宜],美国国家临床生化学院(NACB)发布的室内CV2%,以及根据生物学变异导出的最佳、适当、最低允许不精密度质量规范6种评价标准,计算HbA1c两个批号的室内质控CV和按照仪器系统分组后CV的通过率。结果分别有907家(批号1)和459家(批号2)实验室回报了室内质控数据,根据"指南"中适宜和NACB不精密度要求,满足评价标准的实验室所占比例,批号l为46.8%(当月)、37.0%(累积);批号2为58.0%(当月)、47.1%(累积)。不同仪器系统中生物学变异质量规范能满足的实验室所占比例更低,批号2中Roche组、Primus HPLC组、惠中MQ-2000/2000PT组和Hitachi系列组在适当和最佳标准的实验室满足比例为0%。结论大多数实验室HbA1c的不精密度水平很不理想。     

BIO-RAD D-10血红蛋白检测仪在地中海贫血筛查中的应用

目的:评价BIO-RADD-10血红蛋白(Hb)检测仪在地中海贫血筛查中的应用。方法:对BIO-RADD-10血红蛋白检测仪进行性能评价,并测定我院门诊及住院病人HbA2及HbF含量,同时采用法国Sebia全自动电泳仪进行血红蛋白琼脂糖电泳,比较两种方法对HbA2及HbF含量的检测结果。结果:BIORADD-10血红蛋白检测仪检测HbA2及HbF批内精密度为4.8%、2.77%和1.42%、1.7%,批间精密度为4.91%、3.97%和2.87%、2.73%。共检测了1026例临床标本,并通过全自动琼脂糖凝胶电泳进行定量扫描,得出BIO-RADD-10测定灵敏度为HbF88.3%、HbA297.7%,特异性为HbF96.7%、HbA2为95.6%,阳性预测值为HbF97.4%、HbA296.9%,阴性预测值为HbF85.4%、HbA296.6%。但如果有其他异常血红蛋白条带或血红蛋白H、Bart’s,D-10血红蛋白检测仪不能识别,只能分辨出未知峰。结论:BIO-RADD-10血红蛋白检测仪能够分辨出HbA2及HbF异常增高者,为β地中海贫血的初筛提供快速的诊断依据。     

上海糖化血红蛋白一致性计划回顾分析

目的回顾2010年至2018年参加上海糖化血红蛋白(HbA1c)一致性计划的实验室比对结果,分析各实验室HbA1c检测质量的演变,为进一步提高全国糖化血红蛋白检测能力提供依据。方法回顾性分析。收集2010年至2018年所有参加上海糖化血红蛋白一致性比对计划的实验室比对结果。分析各实验室所在医院分布、检测方法分布的演变。对上海糖化血红蛋白一致性比对计划中各参加实验室历年的判断标准、季度及年度通过率、偏倚、变异系数(CV)进行回顾性分析,并与国家卫健委临检中心、上海市临检中心以及美国病理家协会(CAP)开展的室间质量评价计划结果进行比较。收集2019年第一季度的比对数据,计算各实验室比对结果与靶值的偏倚、变异系数和Sigma(σ)值,并绘制Sigma结合生物医学变异参数评估模型图。结果参加实验室从上海地区9家增长至全国192家,年平均增长率为76.6%;各季度比对判断标准从允许偏差百分比为±8%提高至±6%,各参加实验室通过率从39.1%提高至近90%;各仪器方法组内的最大CV从14.3%降低至4.8%。2019年第一季度,比对结果满足6σ的实验室均近60%,95%以上的实验室满足生物学变异参数模型中的合格及以上标准。结论上海糖化血红蛋白一致性计划使各参加实验室的HbA1c检测结果一致性明显得到了提高。     

三种检测系统测定糖化血红蛋白相关性分析及偏倚评估

目的比较三种检测系统测定糖化血红蛋白（HbA1c）结果的差异并评价其偏倚度。方法分别使用Bio-Rad D-10糖化血红蛋白分析仪应用高压液相层析法（HPLC）、Roche Cobas全自动生化分析仪以免疫法、Afinion AS100分析仪以亲和色谱法测定HbA1c,对64例血液样本作比对测定,评估各检测系统之间测定结果的相关性和偏倚;应用高低两个水平质控品分别测定三种检测系统的不精密度。结果三种不同检测系统测定糖化血红蛋白的结果有良好的相关性（P〈0.01）,HPLC和亲和色谱法的总不精密度〈2.0%,免疫法总不精密度接近4.0%,符合临床测定的性能要求。以D-10 HPLC为参考检测系统,Afinion亲和色谱法和Roche免疫法测定结果的相对偏倚分别为0.17%和0.13%。结论 D-10 HPLC法、Afinion亲和色谱法和Roche免疫法测定糖化血红蛋白的准确度、精密度和偏倚符合临床要求,Roche免疫法测定HbA1c结果变异系数较大,应以HPLC法作为参考检测系统定期作比对分析。     

Evaluation of noninvasive methods for the estimation of haemoglobin content in red blood cell concentrates

Objectives: Red blood cell concentrates (RCCs) are the major blood component transfused to patients. There is a great variability in patient response, depending on both the patient's blood volume and haemoglobin content in the RCC. Standardisation of transfusion practice is needed to improve the prediction of patient outcome. Aim: We hypothesise that labelling of RCCs with haemoglobin content will add possibilities for the standardisation of transfusion practice. Methods: Data from multiple international transfusion services regarding haemoglobin content and weight or volume of RCC were collected and analysed. Results: We demonstrate a strong and highly significant correlation between haemoglobin content with both weight and volume of the RCCs. A linear regression model was used to assess these relationships, and it demonstrates how haemoglobin content can be estimated for different cell production processes. Conclusions: We recommend the use of weight or volume of the RCCs as the basis of estimating haemoglobin in the RCC and postulate that this can be used in future studies to explore the effects of a haemoglobin dose‐based transfusion system. As the weight – and sometimes the volume – of the blood bag is easily accessible, in contrast to direct haemoglobin measurements from each individual unit, this method is feasible and simple.     

高效液相色谱法检测氨甲酰血红蛋白

目的建立一种测定氨甲酰血红蛋白的高效液相色谱法。方法将血红蛋白经酸水解后，释放出氨甲酸缬氨酸，并进一步转化成缬氨酸乙内酰脲后进行定量分析。结果线性范围为０～８０ｍｇ／Ｌ，最低检测量为１４μｇＣＶ／ｇＨｂ平均回收率为７４５７％，批内变异系数（ＣＶ）为８５％，批间ＣＶ为１０９％。并对３０例正常人进行测定。结论该方法快速、灵敏、特异、实用，适于临床常规和研究应用。     

The 'transfusion trigger' in three large West of Scotland hospitals: medical staff questionnaire survey

Efficient use of blood is desirable due to increased demand for blood, shortfall in donations, and the risk of mismatch transfusion accidents and transfusion-transmitted infections. We surveyed transfusion practice in three hospitals. Low haemoglobin was a feature in 91% of transfusion requests and is the commonest 'trigger' for transfusion. Forty-eight per cent of transfusions were initiated with the aim of restoring haemoglobin to 10 g dL(-1). Seventy-three per cent of transfusions were given when the patient's haemoglobin was between 7 and 10 g dL(-1). These figures suggest liberal blood use when compared with international practice. Major healthcare resource and patient safety implications result from this. Evidence-based guidelines issued at both local and national levels would be helpful.     

促红细胞生成素在治疗恶性血液病贫血中应用

目的:探讨促红细胞生成素(EPO)在治疗恶性血液病患者贫血中的临床应用。方法:随机将36例恶性血液病伴有贫血患者分为2组各18例进行对照研究,治疗组18例给予益比奥1万单位皮下注射,每周3次。对照组不用EPO,其他同治疗组。结果:治疗组18例患者治疗前后血红蛋白(Hb)、红细胞压积(HCT)分别为(65.7±9.8)g/L,(0.21±0.03)和(101.2±15.3)g/L,(0.32±0.02)。治疗组治疗4周后Hb浓度较对照组(72.1±12.5)g/L上升明显,P<0.01。结论:应用EPO治疗恶性血液病贫血,不仅能提高Hb水平,而且可以减少输血,副反应少,值得应用于临床。     

Haemoglobin-based erythrocyte transfusion substitutes

Concerns about the infectious and immunosuppressive risks of allogeneic blood products persist, and the increased disproportion of blood donation and consumption has reinforced the search for alternative erythrocyte transfusion strategies in recent years. With the absence of problems such as nephro-toxicity, increased colloid osmotic pressure and sudden renal clearance, modern haemoglobin based oxygen carriers (HBOC) have shown their effectiveness and tolerability in numerous animal and several clinical studies. HBOC can be infused without prior cross-matching and are now available as stable formulations with long shelf-life. Most clinical studies have been performed with human cross-linked haemoglobin (DCLHb) but all trials were stopped two years ago because of an increased mortality in two clinical trials in patients who received DCLHb after stroke and multiple injury shock. However, experimental trials in animals are in progress with DCLHb and recombinant human haemoglobin. In contrast, Phase III studies with polymerised bovine haemoglobin (HBOC-201) are finished or currently under evaluation showing that infusion of HBOC-201 can avoid or reduce allogeneic blood transfusion needs in specific peri-operative settings. As a consequence, HBOC-2001 was actually approved for treatment of peri-operative anaemia in elective adult surgical patients in South Africa. Other human or bovine haemoglobin solutions are currently being investigated in different clinical studies in cardiac surgery patients, sepsis and tumour patients. More recent investigations have shown that HBOC are not only simple erythrocyte transfusion substitutes but highly effective oxygen donators in terms of tissue oxygenation. HBOC open the door for a new therapeutic strategy: plasmatic oxygen delivery with physiological concentrations of inspired oxygen. In specific situations (e.g., ischaemia or arterial stenosis) HBOC have advantages over red blood cells because they can reach post-stenotic or poorly perfused tissues with the plasma stream, where erythrocytes are not able to pass. In addition to significant plasmatic oxygen transport, HBOC also enhance tissue oxygenation because of the facilitated oxygen release by HBOC and from remaining erythrocytes. Further studies will show, if the outcome of patients with impaired perfusion (e.g., stroke or myocardial infarction) can be improved by prophylactic or therapeutic application of HBOC. Whenever these formulations are globally launched, they will find differential indications as potent oxygen-delivering drugs in addition to the globally recognised goal of red cell substitutes in cases of bleeding.     

Tissue oxygenation,strength and lactate response to different blood flow restrictive pressures

This study aimed to determine whether changes in initial restrictive pressures (IRP, tightness of the cuff before inflation with air) affect tissue oxygenation, lactate production and leg strength before, during and after knee extension exercises. The cuff was positioned on the right thigh, and the IRP of either 40–45 or 60–65 mmHg were applied randomly prior to inflating the cuff to the final restrictive pressure (the pressure reached after inflating the cuff with air). Subjects performed four sets (30, 15, 15 and 15 reps) of isotonic knee extensions with 1‐min rest between sets. Tissue oxygenation and blood lactate levels were assessed prior to, during and after exercise, and leg strength was assessed pre‐ and postexercise. There were significant condition by time interactions (P<0·01) and main effects for both condition (P<0·01) and time (P<0·01) for tissue oxygenation, deoxyhaemoglobin, total haemoglobin. Significant main effects were detected for both condition (P<0·01) and time (P<0·01) for leg strength values. There was only a significant time main effect for lactate concentrations. This study is the first to show that a higher IRP had a significant impact on percent tissue oxygenation, leg strength and deoxygenated haemoglobin accumulation during exercise.     

A MALDI-TOF mass spectrometry-based haemoglobin chain quantification method for rapid screen of thalassaemia

BackgroundThalassaemia is one of the most common inherited monogenic diseases worldwide with a heavy global health burden. Considering its high prevalence in low and middle-income countries, a cheap, accurate and high-throughput screening test of thalassaemia prior to a more expensive confirmatory diagnostic test is urgently needed.MethodsIn this study, we constructed a machine learning model based on MALDI-TOF mass spectrometry quantification of haemoglobin chains in blood, and for the first time, evaluated its diagnostic efficacy in 674 thalassaemia (including both asymptomatic carriers and symptomatic patients) and control samples collected in three hospitals. Parameters related to haemoglobin imbalance (α-globin, β-globin, γ-globin, α/β and α-β) were used for feature selection before classification model construction with 8 machine learning methods in cohort 1 and further model efficiency validation in cohort 2.ResultsThe logistic regression model with 5 haemoglobin peak features achieved good classification performance in validation cohort 2 (AUC 0.99, 95% CI 0.98–1, sensitivity 98.7%, specificity 95.5%). Furthermore, the logistic regression model with 6 haemoglobin peak features was also constructed to specifically identify β-thalassaemia (AUC 0.94, 95% CI 0.91–0.97, sensitivity 96.5%, specificity 87.8% in validation cohort 2).ConclusionsFor the first time, we constructed an inexpensive, accurate and high-throughput classification model based on MALDI-TOF mass spectrometry quantification of haemoglobin chains and demonstrated its great potential in rapid screening of thalassaemia in large populations.

Key messages

• Thalassaemia is one of the most common inherited monogenic diseases worldwide with a heavy global health burden.
• We constructed a machine learning model based on MALDI-TOF mass spectrometry quantification of haemoglobin chains to screen for thalassaemia.

     

Changes in Arterial Oxygen Tension Correlate with Changes in End-expiratory Carbon Monoxide Level

Objective Carbon monoxide (CO) and oxygen compete for haemoglobin binding sites. While the effects of increased inspiratory oxygen fractions on exhaled carbon monoxide concentrations have been studied previously, the relationships between intravascular oxygen tension, blood carboxyhaemoglobin levels and expiratory CO concentrations remain unclear. We therefore studied the effects of increases in arterial oxygen tension as crucial determinant for the displacement of carbon monoxide from its haemoglobin bond during lung passage. Methods Measurements of end-expiratory CO concentrations (eCO), arterial oxygen tensions and carboxyhaemoglobin concentrations were performed in 19 patients while breathing air and oxygen. Results With increasing PaO₂ (from 11.5 ± 1.9 to 35.2 ± 10.3 kPa) end-expiratory CO concentrations increased from 8.6 ± 4.9 to 16.7 ± 9.4 ppm, p < 0.001, with a mean increase of 0.36 ppm CO per kPa increase in PaO₂ (ΔeCO [ppm] = 0.36 *␣ΔPaO₂ [kPa]). Increases of arterial oxygen tension correlated with increases of end-expiratory CO concentration (r² = 0.33). Arterial carboxyhaemoglobin concentrations decreased from 1.06 ± 0.37 during air breathing to 0.92 ± 0.35 % after 5 minutes of oxygen inhalation (p < 0.001). Conclusions Oxygen-induced increases in exhaled CO correlate with increases in arterial oxygen tensions. Furthermore, oxygen inhalation reduces carboxyhaemoglobin levels, supporting the concept of accelerated CO elimination by oxygen via the lungs. Patrick Schober, M.D., Melanie Kalmanowicz, Lothar A Schwarte, M.D., PhD, Joerg Weimann, Professor, M.D. and Stephan A Loer, Professor, M.D., PhD, M.Sc. Changes in arterial oxygen tension correlate with changes in end-expiratory carbon monoxide level.     

Influence of D- and L-glucose on erythrocytes and blood viscosity

BACKGROUND: Elevated blood glucose levels are associated with substantial morbidity and mortality. The pathomechanism behind it is not well understood. The aim of the present study was to investigate the effect of glucose on blood rheology. MATERIALS AND METHODS: Blood from healthy volunteers was incubated with various concentrations of D- and L-glucose for 1 h at 37 degrees C. Whole blood viscosity at haematocrit 45% was measured at high and low shear rate (94.5 and 0.1 s(-1)). Erythrocyte shape and volume were assessed. Haemoglobin solutions were incubated with D-glucose for up to 96 h and the viscosity was measured. RESULTS: D-glucose dissolved in H2O and diluted with isotonic NaCl, added to whole blood (additional D-glucose concentrations 0-80 mM), led to a red cell swelling and an increase in blood viscosity at low shear rate (0.1 s(-1)). This process was reversible upon removal of D-glucose. L-glucose, which is not transported into the red cell by the D-glucose-specific transport protein GLUT-1, had no effect. When D-glucose was dissolved and diluted in autologous plasma, haematocrit and viscosity remained unaffected, but L-glucose decreased both values. Incubation of a haemoglobin solution with D-glucose at 37 degrees C led to a time-dependent increase in glycosylated haemoglobin (HbA1C) up to 8%, but left the viscosity unchanged. CONCLUSION: Blood glucose tested in a wide range of concentrations did not affect blood viscosity and morphological or biophysical properties of erythrocytes.