血清总胆红素常规系统测量结果的正确度评价

血清总胆红素(total bilirubin,T-Bil)在临床实验室测量频率较高。目前,我国临床检验领域不同T-Bil测量方法的室间变异可达150%(30μmol/L)[1],给临床诊断和治疗带来很大困惑。理论上讲T-Bil是化学定义明确的小分子,可溯源至SI单位,溯源途径明确。按ISO17511[2]规定,T-Bil测量结果应溯源至T-Bil参考方法[3]。本研究尝试采用"与T-Bil参考方法比较"的方法评价在自动生化分析仪上建立的两种血清T-Bil常规系统测量结果的正确度,现报道如下。     

免疫荧光干式定量法检测可溶性生长刺激表达基因2蛋白性能验证

目的 对免疫荧光干式定量法检测可溶性生长刺激表达基因2蛋白(sST2)进行临床分析性能验证。方法 依据国家卫生行业标准WS/T492-2016及参考美国临床和实验室标准化协会(CLSI)相关文件要求,对实验室免疫荧光干式定量法检测sST2的精密度、正确度、线性范围、参考区间进行评价。结果 sST2的高低两水平的批内精密度分别为3.48%和1.62%,实验室总精密度分别为4.19%和1.47%,两校准品偏倚分别为-1.95%和-0.40%,精密度与正确度均符合行业标准要求;在分析测量范围内线性良好(R²=0.9998),参考区间验证R=100%验证通过。结论 sST2免疫荧光干式定量法的各项性能指标均符合要求,适于临床样本分析。     

Gentier96全自动医用PCR分析系统检测HBV-DNA及HCV-RNA的性能验证

目的 对实验室新购入设备Gentier96全自动医用PCR分析系统使用HBV-DNA及HCV-RNA定量检测项目进行性能验证。方法 通过检测HBV-DNA及HCV-RNA定量检测项目，对精密度、正确度、线性范围及抗干扰能力进行验证和评价。结果 该系统检测低水平和高水平HBV-DNA的实验室变异系数(CV)分别为4.12%、1.27%。低水平和高水平HCV-RNA的实验室变异系数(CV)分别为4.75%、1.46%。HBV-DNA国家标准物质GBW(E)090137(浓度编号S5)靶值的对数值3.15、GBW(E)090139(浓度编号S2)靶值的对数值6.66。HCV-RNA国家标准物质GBW(E)090140(浓度编号S5)靶值的对数值3.34、GBW(E)090142(浓度编号S3)靶值的对数值5.64,实测值与靶值的差异均在±0.4对数值范围内。该分析系统HBV-DNA定量检测在4.69×10～2.04×10⁹范围内呈线性，线性回归方程为Y=0.9884X+0.1452,R²=0.9994。HCV-RNA定量检测在3.47×10～2.3...     

冰冻人血浆醛固酮标准物质在量值传递中的效果评价

目的 评价冰冻人血浆醛固酮国家二级标准物质在临床实验室血浆醛固酮量值传递中的效果。方法 采用同位素稀释液相色谱串联质谱法(ID-LC/MS/MS)和A、B、C 3种免疫法测量原理的常规系统同时测量2个浓度醛固酮标准物质和20份无肉眼可见溶血、乳糜和黄疸的临床单人份血浆样本。以标准物质各常规系统的测量结果为X轴,标准物质的认定值为Y轴作直线回归并计算临床样本经量值递后的值。以ID-LC/MS/MS法测量临床样本结果为X轴,分别以量值传递前后临床样本在A、B、C常规系统与ID-LC/MS/MS法之间结果的相对偏倚为Y轴,绘制改良Bland-Altman图,评价常规系统量值传递前后测量结果的正确性。通过比较量值传递前后平均偏倚及偏倚范围的变化评价醛固酮标准物质在量值传递中的效果。结果 根据正确性评价标准判断,A、B、C常规系统量值传递前后测量结果均不正确。量值传递前临床样本在A、B、C常规系统与ID-LC/MS/MS法之间结果的平均偏倚分别为39.34%、41.45%和97.71%,偏倚波动范围分别为16.93%～95.71%、-3.11%～101.5%和4.55%～319.85%。量值传递...     

基于我国WS/T促甲状腺激素受体抗体检测方法的性能验证

目的 基于我国卫生行业标准(WS/T)对迈瑞高敏磁微粒化学发光法(CLIA)检测TRAb进行性能验证。方法 参照最新WS/T文件,以Roche Cobase 601电化学发光法(ECLIA)作为已验证的检测方法,对迈瑞CLIA的精密度、基于患者血清的正确度、线性、检出限、参考区间进行验证,评价迈瑞与Roche的诊断符合率。结果 迈瑞CLIA检测TRAb的批内CV为1.56%～4.60%,实验室内CV为1.65%～5.19%;与Roche ECLIA的实验室相对差值为-3.6%;在0.02～44.00IU/L范围内线性拟合方程Y=1.040X+0.636,线性系数R²=0.990;24个检出限临界值数据22个(91.7%)≤0.3IU/L;20个健康体检者血清TRAb检测结果仅1个超出参考区间≤1.9IU/L;与Roche ECLIA的诊断总符合率90%,阳性符合率75%,阴性符合率100%,Kappa检验值0.7826(P<0.001)。结论 迈瑞高敏磁微粒CLIA的精密度、基于患者血清的正确度、线性范围、检出限均符合厂家声明,参考区间经验证可转移至我实验室...     

冰冻人血清胱抑素C标准物质的制备

目的 研制人血清基质的胱抑素C标准物质。方法 收集无溶血、黄疸、乳糜的血清样本,去除血清中的不稳定成分,经过浓度配比、过滤、混匀、分装后制成4个浓度的胱抑素C标准物质。在单一实验室内采用10种免疫透射比浊法测量原理的常规方法同时测量系列浓度ERM-DA471/IFCC溶液和各标准物质中胱抑素C浓度,以经过ERM-DA471/IFCC量值传递后的胱抑素C浓度均值作为标准物质的认定值。依据文件分别评估胱抑素C标准物质的均匀性、稳定性及互换性。结果4个水平的胱抑素C标准物质定值结果为0.89±0.10mg/L、1.82±0.13mg/L、3.55±0.28mg/L、5.25±0.29mg/L。制备的标准物质均匀性良好。-70℃以下条件可稳定半年,2～8℃条件可稳定144h,复融后室温条件可稳定8h。常规测量系统中的互换性良好。结论 该标准物质符合国家二级标准物质的要求,现已通过全国标准物质管理委员会的审批,编号为GBW(E)091173-091176,可用于临床实验室胱抑素C的质量评价。     

免疫透射比浊法血清白蛋白检测试剂盒的性能验证和临床应用评价

目的对免疫透射比浊法血清白蛋白检测试剂盒进行性能验证和临床应用评价。方法根据美国临床实验室标准化委员会(CLSI)的标准要求,使用全自动日立生化分析仪7600-020,对该试剂盒的精密度、正确度、检出限、线性范围、干扰实验、参考区间、方法学比对进行验证,并将检测结果和厂家提供的的性能指标进行比较。结果该试剂检测血清白蛋白的批内精密度为1.1%、批间精密度为1.2%,均小于厂家提供的指标;正确度在允许偏倚范围内;检出限为0.152g/L;线性斜率为0.9755,相关系数的平方R2为0.9945,大于0.99符合要求;生物参考区间符合率100%;在厂家说明书标示的干扰物浓度范围内,对检测结果没有明显影响;与溴甲酚绿法做比对实验,相关性回归方程为Y=0.9894X-1.1033,R2为0.9794,在低值结果中两种方法学差异呈非线性分布,无法用校正系数调整。结论免疫透射比浊法血清白蛋白检测试剂盒在7600-020生化分析仪上主要分析性能符合厂家的声明,结果准确可靠,抗干扰能力强,可以开展用于临床检验分析。     

Waters Xevo TQD串联质谱仪筛查新生儿遗传代谢病的性能验证

目的对Waters公司XevoTQD串联质谱仪及其配套试剂PerkinElmer公司NeoBaseTMNon-derivatizedMSMS kit在测定新生儿多种遗传代谢病筛查的指标-血氨基酸和肉碱的能力进行性能验证。方法参考《中华人民共和国卫生行业标准》WS/T 420-2013中临床实验室对商品定量试剂盒分析性能的验证,依据CLIA88要求,对精密度和正确度两项主要性能进行验证。结果 (1)XevoTQD检测9种氨基酸的批内和批间精密度分别为2.69%-5.31%和3.96%-10.28%,检测13种肉碱的批内和批间精密度分别为2.59%～7.84%和4.03%～11.51%。(2)XevoTQD检测多种氨基酸和肉碱的正确度均符合要求;结论 Waters Xevo TQD检测氨基酸及肉碱的性能通过验证,满足实验室要求。     

全自动生化分析仪检测肌酸激酶同工酶质量分析性能验证

目的验证全自动生化分析仪定量检测系统检测肌酸激酶同工酶质量(CKMB mass)的分析性能,确保检验结果的准确。方法参照CLSI系列相关文件、国家相关卫生行业标准、并结合实际情况,对定量检测系统的精密度、正确度、线性范围、临床可报告范围、生物参考区间等分析性能进行验证、评价,将实验室验证结果与质量允许范围进行比较;并对CKMB/CK活性升高或倒置现象进行验证。结果贝克曼AU5400全自动生化分析仪测定CKMB mass浓度在14.185、36.467 ng/mL时,实验室批内不精密度(CVr)分别为2.778%、2.172%,总不精密度(CVl)分别为4.507%、2.255%;与罗氏E602全自动电化学发光分析仪测定CKMB mass进行患者样本比对,回归方程Y=1.0277 X-0.5778,R2=0.9846,相关性良好,5、10、15、90 ng/mL浓度水平处的预期偏倚分别为-8.8%、-3.0%、-1.1%、2.1%,可接受;线性范围为1.865~186.34 ng/mL;临床可报告范围为1.865~1490.72 ng/mL;生物参考区间0~5 ng/mL适用于本实验室;能够很好的纠正实验室CKMB/CK活性假性升高或倒置的现象。结论贝克曼AU5400全自动生化分析仪测定CKMB mass的精密度、正确度、线性范围、临床可报告范围、参考区间等性能良好,能够规范检测、确保检测质量,满足临床需求。     

Lp- PLA2酶法测定试剂盒的临床应用评价

目的 验证北京九强金斯尔脂蛋白相关磷脂酶A2(Lp-PLA2)酶法测定试剂盒的性能指标，评价其是否符合临床需求。方法 根据WS/T492-2016《临床检测定量测定项目精密度与正确度性能验证》、CNAS-GL037:2019《临床化学定量检验程序性能验证指南》及WST402-2012《临床实验室检验项目参考区间的制定》等文件要求，对北京九强金斯尔Lp-PLA2酶法测定试剂盒的正确度、精密度、线性范围、可报告范围和生物参考区间等性能进行验证，并与上海润鸿Lp-PLA2测定试剂盒进行方法学比对。结果 正确度验证中，金斯尔Lp-PLA2酶法测定试剂盒测定厂家工作标准品偏差均小于8%;检测配套高低两水平质控品，每个水平检测的重复性和实验室内精密度的变异系数(CV)均在±2%范围内；在线性区间评价试验中，得到线性回归方程Y=1.0054X-3.3466,R²=0.9998。验证线性区间为58.53～1 181.07U/L,最大可稀释倍数为64倍，临床可报告上限水平为75 588.48U/L。厂家提供的生物参考区间男性：230～728U/L,女性：194～640U/L(18～...     

国产胰岛素放射免疫检测系统的现状与分析

目的调查研究目前国内胰岛素(INS)放射免疫检测系统的现状。方法首先收集门诊及住院患者血清标本140份,选用E170电化学发光免疫分析仪和四种国产INS放射免疫诊断试剂盒(国产a、b、c、d)对上述血清进行检测,根据E170检测的血清INS水平特点,把检测数据分为四组;其次,人工配制低、高浓度INS混合血清2份,分别用国产放免诊断试剂盒重复检测20次并计算批内变异系数(CV%);另外又收集E170检测血清INS≤10μIU/mL血清68份,用国产a试剂盒检测,然后对二者结果进行回归分析,所得回归方程用来校正国产a水平1组INS数据,再将校正后数据与E170数据做配对t检验以及相关性分析。结果 (1)对四种国产放射免疫诊断试剂盒INS检测结果进行方差分析,四个水平组的F值分别是46.7、13.8、9.6、9.2,P值均小于0.001。(2)四种国产试剂低和高浓度混合血清批内CV%均控制在10%以内。(3)与E170检测结果(5.4±2.5)μIU/mL相比,校正后国产a试剂检测水平1组血清INS含量为(5.6±1.6)μIU/mL,二者相比无统计学差异(t=0.49,P0.05),校正前后相关系数r均为0.49。结论国产胰岛素放射免疫检测系统间缺乏可比性和溯源性,需要建立一个完善的INS参考系统来提高检测系统的溯源性,以保证检测结果的准确度。     

HBV-DNA阳性患者血清CG水平测定的临床价值探讨

目的:探讨HBV-DNA阳性患者血清甘胆酸(CG)水平测定的临床意义。方法:以PCR法测定的196例HBV-DNA阳性患者作为感染组,120例健康体检者作为对照组,检测两组空腹血清CG和ALT、T-Bil的水平(CG用RIA,ALT及T-Bil用生化法)。结果:血清CG、ALT水平与对照组比较均显著升高(P<0.01),T-Bil无显著性差异(P>0.05),CG的阳性检出率高达76.0%(149/196)。结论:血清CG测定对HBV-DNA阳性患者具有重要的临床意义,优于常规肝功能检查。     

Effect of menopause on low-density lipoprotein oxidation: is oestrogen an important determinant?

Objectives: Significantly increased risk for developing cardiovascular disease in post-menopausal women is linked with the fall of oestrogen. Although supraphysiological levels of oestrogen may inhibit oxygen free radical mediated low-density lipoprotein (LDL) oxidation, the effect of physiological level of oestrogen on LDL oxidation is unknown. Methods: The present study compared oxidizability of LDL in healthy pre- and post-menopausal women by using a commonly employed copper ion-dependent method. Results: Pre-menopausal women (n=20, mean age 27) had significantly higher serum oestradiol level (576±109 pmol/l) in comparison to post-menopausal women (n=23, mean age 51, oestradiol 64±18 pmol/l, P<0.001). The oxidation of LDL in two groups was not different by measuring either the lag phase of conjugated dienes formation (54±12 vs. 55±14 min, P>0.05) or the generation of thiobarbituric acid reactive substances over 4 h of oxidation. The major lipid soluble antioxidant in LDL, vitamin E (determined as -tocopherol) is similar in two groups (2.34±0.48 vs. 2.40±0.56 nmol/mg LDL, pre- and post-menopausal subjects, respectively, P>0.05). Linear regression analysis found a weak but significant correlation between LDL vitamin E level and oxidizability of LDL in both groups but did not show effect of serum oestradiol levels. Conclusion: The results suggest that physiological levels of oestrogen may not be able to affect in vitro LDL oxidation.     

面部特征及软组织测量方法的研究进展

面部特征在体质人类学、法医学及美容整形研究中是一项重要内容,国内外专家做了深入的研究.软组织测量技术在颅骨面貌复原中发挥着重要作用,要尽可能地恢复原貌,就必须掌握面部各个关键位点软组织的厚度;常用的测量方法有:照片测量法,方法简便实用、成本低,但不够准确;X线测量法,测量结果可靠为大多数学者所接受,但使用不方便;而三维...     

A Valuable Antigen Detection Method for Diagnosis of Acute Hepatitis E

Hepatitis E virus (HEV) is a serious public health problem. The commonly used tests that are specific for current HEV infection diagnosis include the detection of anti-HEV IgM and HEV RNA. Here, we report an improved enzyme-linked immunosorbent assay (ELISA) method for HEV antigen detection with a linear range equivalent to 6.3 × 10³ to 9.2 × 10⁵ RNA copies per ml. The monoclonal antibody (MAb) 12F12, a high-ability MAb that binds HEV virus, was selected as the capture antibody from a panel of 95 MAbs. The positive period of HEV antigenemia in infected monkeys using this test was, on average, 3 weeks longer than previously reported and covered the majority of the acute phase. The positive detection rates of IgM, RNA, and new antigen from the first serum samples collected from 16 confirmed acute hepatitis E patients were 81% (13/16), 81% (13/16), and 100% (16/16), respectively. In three patients, the initial serum specimens that tested negative for IgM, despite the presence of symptoms of acute hepatitis and elevated alanine aminotransferase (ALT) levels, were positive for HEV antigen and HEV RNA. In contrast, the serum samples of the three RNA-negative patients were antigen positive (and IgM positive), possibly due to the degradation of HEV nucleic acids. Our results suggest that this new antigen detection method has acceptable concordance with RNA detection and could serve as an important tool for diagnosing acute hepatitis E.     

BA-ELISA检测谷胱甘胱转移酶对肝癌的诊断意义

应用抗总的谷胱甘肽转移酶(EC 2.5.1.18 GSTs)的抗体,建立了生物素-亲和素-酶联免疫吸附法(BA-ELISA),并用以检测肿瘤病人血清中总GSTs浓度.本法非常敏感,其最低检出浓度可达0.05ng/ml,在0.05～6.0ng/ml的范围内,血清中总GSTs浓度和其相应之光密度值呈很好的直线关系.批内和批间的变异系数分别为8.2％和12.0％.肝癌、肝炎、消化道肿瘤、良性疾病患者血清总GSTs浓度分别为3.29±2.64,0.83±0.71,0.57±0.47和0.58±0.51ng/ml(mean±SD),而正常对照组仅为0.33±0.29ng/ml.肝癌病人血清总GSTs浓度显著高于其他疾病患者和正常人,总GSTs可望成为肝癌诊断的一个肿瘤标志。     

Clinical significance of determination of serum RNase activities in patients with eosinophilia--II. Measurements using polyuridylic acid and polycytidylic acid as substrates

The activities of RNase (RNase-U and RNase-C) were determined in the serum and leukocytes of 277 patients with 14 cases of various kinds of eosinophilia (not less than 10(3)/microliters), 28 cases of chronic myelocytic leukemia (CML), using polyuridylic acid and polycytidylic acid as synthetic substrates according to the method of Raddi et al. Serum RNase-U activity, serum RNase-C activity and the activity ratio (U/C x 10(-3)) were 55 +/- 14 U, 1,280 +/- 235 U and 44 +/- 11 (mean +/- SD), 196 +/- 137, 1,992 +/- 1,134 U and 97 +/- 38, and 110 +/- 50 U, 1,854 +/- 625 U and 65 +/- 13 for normal subjects, eosinophilia and CML (untreated), respectively. U/C ratio in eosinophilia and CML (untreated) showed a highly significant positive correlation (p less than 0.001) with peripheral eosinophil count; the activity of serum RNase-U per cells in the supernatant of eosinophil homogenate rose significantly (p less than 0.001) compared with that of lymphocytes or granulocytes. Besides, serum and eosinophil RNase-U had a similar optimal pH. These results suggested that serum RNase-U in eosinophilia originated mostly from eosinophils and its rise was correlated strongly with the increase in eosinophils.     

An avidin-biotin ELISA for the measurement of mitochondrial aspartate aminotransferase in human serum

We describe a new double-antibody ELISA system using avidin-biotin amplication for the mass measurement of mitochondrial aspartate aminotransferase (m-AST) in human serum. The assay is very sensitive and as little as 0.5 ng/ml of m-AST may be detected. The method is linear up to 100 micrograms/l. The within-day and day-to-day coefficients of variation were found to be 8.9% and 11.5% respectively for a low m-AST concentration (2.7 micrograms/l), and 5.9% and 8.0% respectively for a high level of m-AST (30 micrograms/l). The assay requires 100 microliters of serum and can be completed within 5 h. The ELISA procedure and a classical immunoprecipitation technique measuring the catalytic activity of the isoenzyme were applied simultaneously to the sera of 189 subjects. The protein levels determined by ELISA correlated poorly (r = 0.66) with the catalytic activity of m-AST.     

国产甲磺酸培氟沙星体内抗菌活性试验

口服和静脉给予国产甲磺酸培氟沙星对大肠杆菌-12、克氏肺炎杆菌-64和绿脓杆菌-14感染小鼠均有保护作用,其中口服给药的PD_(50)分别为1.08,5.87和8.10mg/kg;静脉给药的PD_(50)分别为0.54,3.10和7.02mg/kg。