酶法测定二氧化碳的使用评价

目的　对液体双试剂酶终点法检测血清中的二氧化碳 ( CO2 )试剂进行应用评价。方法　通过精密度试验、线性试验、对比试验和反应动力学曲线观察。结果　精密度测定结果批内 CV为 1.2 6%～ 1.61% ,批间 CV为 3 .71%～ 7.5 2 %。线性试验结果相关系数为 0 .9998,线性达 5 0 m mol/ L。与经典电极法对比测定结果相关系数为 0 .92。结论　该法简便、快速准确 ,用血量少 ,适用于自动分析仪 ,有推广应用价值     

ISO15189体系下全自动血凝分析仪的3Q验证

目的以SysmexCS-2500全自动血凝分析仪为例,探究医学实验室质量和能力认可准则体系(ISO15189)下全自动血凝分析仪的3Q验证过程。方法严格按照ISO15189的要求,对SysmexCS-2500全自动血凝分析仪器的凝血酶原时间(PT)、活化部分凝血活酶时间(APTT)、凝血酶时间(TT)、纤维蛋白原(FIB)、D-二聚体五个指标进行批内精密度、日间精密度、准确性、线性范围、检测下限(灵敏度)、携带污染率性能验证。结果批内精密度:PT、APTT、TT、FIB、D-二聚体的正常质控CV值分别为0.34%、0.38%、0.62%、1.04%、3.32%;PT、APTT、FIB、D-二聚体的异常质控CV值分别为0.47%、0.41%、2.56%、9.70%;日间精密度:PT、APTT、TT、FIB、D-二聚体的正常质控CV值分别为2.81%、2.15%、3.72%、3.51%、5.67%;PT、APTT、FIB、D-二聚体异常质控CV值分别为2.21%、2.77%、4.40%、5.37%;准确性:PT、APTT、FIB、TT正常质控水平测试结果分别为12.1、25.6、2.71、19.7;PT与FIB异常质控水平测试结果为19.9、0.89;D-二聚体低值重复三次测试结果分别为0.28、0.29、0.29;D-二聚体高值重复三次测试结果分别为2.20、2.22、2.21;FIB与D-二聚体线性范围验证结果 a值分别为0.9535、1.0162,R2分别为0.9911、0.9975;灵敏度:D-二聚体与FIB的CV值分别为3.48%与2.57%;携带污染率最高值为2.65%。所有验证结果均符合要求,验证合格。结论通过ISO15189体系下的3Q验证,SysmexCS-2500全自动血凝分析仪的主要性能均符合要求,可用于临床凝血检测等工作。     

Access2免疫化学发光仪检测动物血清心肌钙蛋白I的分析性能确证

目的对美国贝克曼库尔特有限公司Access2免疫化学发光仪的主要分析性能进行验证。方法对该仪器检测项目血清心肌钙蛋白I(Cardiac Troponin I,c Tn I)进行性能验证。利用变异系数分别对质控及大鼠、犬和食蟹猴血清心肌钙蛋白I的精密度进行评估,并根据质控精密度数据对质控的靶值范围进行计算。利用相对偏差及靶值范围对准确度进行评价;利用携带污染率对携带污染情况进行评价。利用线性方程对Trop I的线性范围进行评价,同时对Access 2免疫化学发光仪检测动物血清c Tn I的功能灵敏度进行了评价。结果 Access2免疫化学发光仪检测c Tn I低、中、高3个水平质控的批内RSD均<8%,批间RSD均<63.75%,大鼠、犬和食蟹猴血清c Tn I批内RSD均<8%。Bio-Rad 3水平质控的靶值范围分别为:0.363 8~0.473 5、1.898 6~2.819 3、7.950 3~9.367 6 ng/m L。Access2免疫化学发光仪检测c Tn I的RSD<16.32%,且低、中、高3水平质控均在各自的靶值±30%范围内。携带污染率<10%,线性验证高值动物血清按一定比例稀释后将所得理论值与实测值进行回归分析,a值介于0.95~1.05,R>0.975,线性范围为0.016 0~82.843 3 ng/m L,功能灵敏度为0.007 6 ng/m L。结论 Access2免疫化学发光仪检测c Tn I的精密度、准确度、携带污染率及线性范围均在可接受范围内,且本研究获得了动物血清c Tn I检测的线性范围及功能灵敏度。因此,Access2免疫化学发光仪可用于临床前不同动物种属c Tn I的检测。     

NM-BAPTA法钙检测试剂盒的性能评价

目的 对NM-BAPTA法钙检测试剂盒进行性能评价.方法 根据美国临床与实验室协会(CLSI)指南文件EP15-A2、EP9-A2、EP6-A、C28-A2,验证日立7180生化分析仪上NM-BAPTA法钙检测试剂盒的精密度、准确度、线性范围、生物参考区间.结果 高低2个浓度的血钙质控品批内精密度标准差(SD)分别为0.03 mmol/L、0.02 mmol/L,批间精密度标准差分别为0.08 mmol/L、0.06 mmol/L;准确度验证其血钙结果与参比系统cobas 8000血钙结果呈良好的相关性,r2=0.954 8,相对偏差＜12.5％;线性范围验证斜率为0.971,r2=0.999,线性范围为1.00～4.70 mmol/L,线性理想;参考区间验证符合率100％.结论 NM-BAPTA法钙检测试剂盒在日立7180生化分析仪上的分析性能符合质量目标要求,可应用于临床检测.     

日立7080型自动生化分析仪临床应用评价

目的 对日立7080型自动生化分析仪进行应用评价。方法 采用日立公司提供的ROCHE试剂,ROCHE定值质控血清批号:151342010,病人血清,正常质控血清和病理质控血清,连续测试10次,对该仪器进行精密度和准确度测试。结果 仪器精密度试验,病人血清测试的CV值在0.08%～1.80%之间,正常质控血清CV值在0.04%～2.20%之间,病理质控血清CV值在0.04%～0.80%之间。仪器准确度试验,ISE准确度系数均在99%以上,肾功能项目的准确度系数均在97%以上,肝功能项目准确度系数均为95%以上,结论 该议器主要指标评价结果符合设计性能,适用各级医院检验科及实验室生化分析[1]。     

Ferene-s络合法检测血清铁的方法学评价

目的:探讨对Ferene-s络合法检测血清铁进行方法学评价。方法:用Ferene-s络合Fe2 生成红蓝色复合物测其终点吸光度的变化,观察其显色稳定性、灵敏度、线性、精密度、准确度、溶血干扰。结果:显色物最大吸收波长566nm,显色后5～60分钟内稳定,灵敏度1.875μmol/L,线性范围0～120μmol/L,批内CV值1.57%,批间2.40%,回收率103.5%,标本中血红蛋白含量>1.5g/L对测定有影响。结论:该方法显色稳定、灵敏度高、线性好范围宽、精密度好、准确度高、血清用量少,具有操作简单、快速等特点,适用于临床检验室采用。     

Access 2免疫化学发光仪检测动物血清心肌钙蛋白I的分析性能确证

目的 对美国贝克曼库尔特有限公司Access2免疫化学发光仪的主要分析性能进行验证。方法 对该仪器检测项目血清心肌钙蛋白I(Cardiac Troponin I, cTnI)进行性能验证。利用变异系数分别对质控及大鼠、犬和食蟹猴血清心肌钙蛋白I的精密度进行评估, 并根据质控精密度数据对质控的靶值范围进行计算。利用相对偏差及靶值范围对准确度进行评价;利用携带污染率对携带污染情况进行评价。利用线性方程对TropI的线性范围进行评价, 同时对Access 2免疫化学发光仪检测动物血清cTnI的功能灵敏度进行了评价。结果 Access2免疫化学发光仪检测cTnI低、中、高3个水平质控的批内RSD均<8%, 批间RSD均<63.75%, 大鼠、犬和食蟹猴血清cTnI批内RSD均<8%。Bio-Rad 3水平质控的靶值范围分别为:0.363 8～0.473 5、1.898 6～2.819 3、7.950 3～9.367 6 ng/mL。Access2免疫化学发光仪检测cTnI的RSD<16.32%, 且低、中、高3水平质控均在各自的靶值±30%范围内。携带污染率<10%, 线性验证高值动物血清按一定比例稀释后将所得理论值与实测值进行回归分析, a值介于0.95～1.05, R> 0.975, 线性范围为0.016 0～82.843 3 ng/mL, 功能灵敏度为0.007 6 ng/mL。结论 Access2免疫化学发光仪检测cTnI的精密度、准确度、携带污染率及线性范围均在可接受范围内, 且本研究获得了动物血清cTnI检测的线性范围及功能灵敏度。因此, Access2免疫化学发光仪可用于临床前不同动物种属cTnI的检测。     

免疫透射比浊法β2-微球蛋白试剂盒的性能评价

目的 了解免疫透射比浊法β2-微球蛋白试剂盒的性能指标。方法 测定该试剂盒的检测下限、精密度、准确度、线性范围。结果 免疫透射比浊法β2-微球蛋白试剂盒的检测下限为0．09mg／L，批内CV小于4％，批间CV小于8％，准确度为99．29％，线性范围为0．2～18mg／L。结论 免疫透射比浊法β2-微球蛋白试剂盒可以满足临床检测的需要。     

紫外分光光度法检测血管紧张素转移酶的方法学评价

目的:评价北京九强公司生产的血管紧张素转移酶(angiotensin-converting enzyme,ACE)检测试剂盒的方法学,分析试剂盒的线性,偏差,携带污染.方法:依据临床和实验室标准协会颁布的<定量临床检验方法的初步评价,批准指南第二版(EP10-A2)>提供的方法,按特定的顺序连续测定低、中、高浓度的质控品5天,从而对该试剂盒检测方法进行初步评价.结果:当ACE低、中、高浓度的质控血清的靶值分别为49、89.5、130 U/L时,按特定的顺序连续测定5天,没有离群值.线性回归方程为y=0.8791x+8.4476,相关系数为0.99637;绝对偏差分别是0.64、1.26、-9.6 U/L;总不精密度(CV%)分别为4.39%、2.26%、1.39%.结论:ACE检测试剂盒检测方法的线性、总变异和抗交叉污染能力均达到EP10-A2文件的标准,符合临床应用要求,高值的偏差超出可接受范围.     

两种方法检测血清甲胎蛋白(AFP)的比较

目的对电化学发光免疫法与微粒子酶联免疫法检测血清甲胎蛋白作对比分析.方法随机抽取150份临床送检的标本,用电化学发光免疫法与微粒子酶联免疫法测试甲胎蛋白的含量,对测试数据进行相关性试验、线性试验、精密度试验、回收试验.结果两法无显著性差异(P＞0.05),但电化学发光免疫法的线性范围较宽(1-980ng/ml),重复性较好(批内CV值3.4%,批间CV值3.5%),回收率较高(97.2%).结论电化学发光免疫法检测血清甲胎蛋白优于微粒子酶联免疫法检测血清甲胎蛋白     

A simple method for quantifying fentanyl,sufentanil, or morphine in discard syringes from anesthesia procedures

Fentanyl, sufentanil, and morphine are commonly used in the conduct of anesthesia. Medical staff working with these drugs are at high risk of addiction. To detect and prevent diversion, a method was developed to quantify these drugs in discard syringes using the BioRad REMEDi HS Drug Profiling System. For fentanyl, the lowest concentration detected is 0.1 microg/mL, and the assay is linear to 5.0 microg/mL; the within-run coefficient of variation (CV) is 0.9% (n = 5), and between-run CV is 2.5% (n = 20). For sufentanil, the lowest concentration detected is 0.5 microg/mL, and the assay is linear to 11.0 microg/mL; the within-run CV is 2.0% (n = 5), and the between-run CV is 2.4% (n = 20). For morphine, the lowest concentration detected is 0.5 microg/mL, and the assay is linear to 10.0 microg/mL; the within-run CV is 11.6% (n = 5), and between-run CV is 11.3% (n = 20). Other drugs commonly used in the operating room were checked for cross-reactivity on the REMEDi HS; none cross-reacted. The REMEDi HS can be used for rapid, accurate quantification of fentanyl, sufentanil, and morphine in discard syringes from anesthesia procedures or related medical applications.     

Roche Modular PPI全自动生化分析仪测定Glu的性能评价

目的评价罗氏Modular PPI全自动生化分析仪测定Glu的基本工作性能。方法对同一批号正常水平、病理水平质控血清在同一天内随标本平行测定20次,计算批内不精密鹿每天各测定1次,测定20天,计算批间不精密度。根据卫生部2010年室间质评的数据及批内、批间不精密度,计算出扩展不准确度。通过对高浓度样品按一定比例稀释,测定样品中Glu的含量,验证厂家的线性范围。结果正常水平、病理水平质控血清批内不精密度分别为0．49％、0．24％;批间不精密度分别为0．58％、0．27％;不准确度分别为±3．1％、±2．2％。线性范围为0～32mmol／L。结论通过对不精密度、不准确度和线性范围实验观察,认为罗氏Modular PPI全自动生化分析仪Glu测定的性能可以满足临床化学实验室的要求。     

葡萄糖-6-磷酸脱氢酶缺乏症基因诊断新技术研究

目的：研究分析用于葡萄糖-6-磷酸脱氢酶（ glucose-6phosphate dehydrogenase ,G6PD）缺乏症基因诊断的多重SNaPshot基因诊断技术这一新技术的检出效果,以便于有效地进行新生儿筛查,和疾病的预防。方法选取2014年7月至2015年5月既往有G6PD生育史,或孕前筛查、新生儿筛查发现的该病患者和G6PD缺乏症高风险胎儿156例为研究对象,并将研究对象随机分为观察组和对照组,每组78例。对照组采用传统的基因检测技术G6PD／6-磷酸葡萄糖酸脱氢酶（6-phosphogluconate dehydrogenase ,6PGD）比值法,观察组则采用新的基因诊断技术-多重SNaP-shot基因诊断,分别检测对G6PD和编码该酶的基因诊断,以基因测序结果为“金标准”对比2组的检出结果。结果观察组待检者检出57例患有由基因突变导致的G6 PD缺乏症,总异常率为73?．08%（57／78）,包括错义突变的A95 G型、G392T型、G1376T型、G1388A 型分别为5例、3例、17例、27例,其发生率分别为6．41%、3．85%、21．79%、34．62%,同义突变5例,发生率为6．41%,男、女异常率不同,女性均为杂合子;G6PD／6PGD比值法测定的78例待检者血样中总共有52例比值结果<1．0,总阳性率为66．67%（52／78）,其中男性、女性的阳性例数分别为30、22;观察组采用的SNaPshot基因诊断技术,测出的突变类型及每种类型的突变数均与测序结果完全一致,而对照组采用的G6PD／6PGD比值法测定酶活性,只能测定错义突变,且存在漏诊,准确率为96．3%,无法检出同义突变。结论SNaPshot基因诊断技术可以有效地检出是那种基因突变导致G6PD缺乏,准确率高,有较高的临床推广应用价值。     

AN IMPROVED KINETIC METHOD FOR MEASUREMENT OF URINARY ALANINE AMINOPEPTIDASE (AAP) ACTIVITY AS A NEPHROTOXIC MARKER

This study examines the effect of various factors that influence alanine aminopeptidase (EC 3.4.11.2; AAP) activity in untreated urine samples, using L-alanine- p -nitroanilide as substrate. An improved kinetic method for assay of AAP activity without pretreatment of urine samples was obtained. The results show that the final concentration of 6 mmol/ L L-alanine- p -nitroanilide and 325 mmol/ L tris(hydroxymethyl)aminomethane (pH 7.90 at 30 C) and the sample/ final reaction volume ratio of 1/ 25 to be optimum. The within-run coefficient of variation (CV) was 1.40%; the between-run CV averaged 3.11%. Reference values were established for AAP activity of untimed urine samples obtained from 810 healthy persons. The normal reference interval (mean 1 SD) was 0.81-38.64 (12.76 6.88) U/ g of creatinine. Freezing the untreated urine sample resulted in a considerable loss of AAP activity. However, when urinary AAP activity is measured by the present method, there is no need for urine and reagent blanks; assay of the enzyme in urine is possible using one reagent solution. Therefore, the present method is suitable for use with various automated analyzers. Finally, the present method requires only 30muL of untreated urine samples, so it is suitable for evaluating of nephrotoxicity in human (especially neonates and infants) and small animal models such as rats.     

Measurement of cyclosporine in plasma of cardiac allograft recipients by fluorescence polarization immunoassay

The Abbott TDx fluorescence polarization immunoassay (FPIA) procedure for measuring cyclosporine A (CsA) was evaluated and compared with the Sandoz polyclonal radioimmunoassay (CsA RIA kit) method. This drug assay was evaluated for precision, calibration, stability, and accuracy. Within-run precision studies utilizing 25 replicate analyses of the three control preparations (containing CsA in the 60-800 ng/ml range) resulted in coefficients of variation (CV) ranging from 1.0 to 9.1%. The CVs of between-run precision determined by assaying the same control drug levels for five consecutive working days ranged from 3.9 to 4.6%. Calibration curve stability was assessed by examining the drift in control values over a 2-week period. Maximum plasma ranged from 82.6 to 108.2%. Four hundred plasma samples were obtained from 30 heart-transplant patients during the first 6 months of CsA therapy and each sample was analyzed simultaneously by TDx and RIA. Linear regression analysis of the results obtained for each patient (x = RIA, y = FPIA) revealed the following mean values: r = 0.87, (CV = 13.7%), slope = 1.47 (CV = 39.2%). Moreover, the concentration of CsA was determined in 35 patient samples both by TDx and high-performance liquid chromatography (HPLC). FPIA results up to 12 times higher than HPLC results have been noted.     

应用NCCLS EP5-A方案评价唾液酸试剂盒的精密度

目的 评价测定唾液酸(SA)试剂盒的精密度.方法 利用NCCLS EP5-A方案评价唾液酸的精密度.结果 正常和异常血清的批内变异系数分别为0.9%和1.9%.批间变异系数分别为1.2%和1·7%,总变异系数为1.6%和2.6%.结论 SA试剂盒精密度良好,准确可靠且操作简便.     

Development and validation of an LC-MS-MS method for the determination of terfenadine in human plasma

A sensitive LC-MS-MS method capable of quantifying terfenadine at levels down to 100 pg ml⁻¹ in human plasma is reported. The method was validated over a linear range from 0.1 to 5.0 ng ml⁻¹ using a liquid-liquid extraction with a deuterium-labelled internal standard. The between-run precision and accuracy of the calibration standards were 2.6–6.0% RSD and −2.0 to +2.2% relative error (RE). The between-run and within-run precision and accuracy of quality control samples (0.3, 1.5 and 3.5 ng ml⁻¹) were 1.0–5.9% RSD and +1.7 to +6.3% RE. This method was applied to the analysis of human plasma samples.     

Use of the EMITtox serum tricyclic antidepressant assay for the analysis of urine samples

The EMITtox serum tricyclic antidepressant (TCA) assay is intended for use with serum or plasma. Currently, there are no commercially available immunoassay kits available for the detection of TCAs in urine. The proposed method utilizes the EMITtox serum TCA assay for the direct analysis of urine samples. The minimum detectable concentration of nortriptyline in urine is 25 ng/mL. The within-run CV was determined to be 0.6% and the between-run CV was 2.6%. The TCA assay cross-reacts with phenothiazines and antihistamines. The proposed methodology should be applicable to other EMIT serum assays to allow their use with urine.     

Evaluation of a new phencyclidine enzyme immunoassay for the detection of phencyclidine in urine with confirmation by high-performance liquid chromatography-tandem mass spectrometry

We present an evaluation of a new phencyclidine (PCP) enzyme immunoassay (PCPI) for detection in urine. The PCPI was evaluated by testing 523 urine specimens. Controls containing 0 ng/mL of PCP and -25% (negative control) and +25% (positive control) of the 25 ng/mL cutoff calibrator were analyzed with each batch. All urines were analyzed by high-performance liquid chromatography- tandem mass spectrometry (HPLC-MS-MS) for PCP. Of the 523 specimens tested, 218 yielded positive results by the PCP assay. HPLC-MS-MS confirmed the presence of at least 25 ng/mL in 214 specimens, indicating four false-positive results containing 11, 13, 16, and 20 ng/mL PCP. Three specimens yielded a negative result; however, PCP concentrations were within 20% of the 25 ng/mL cutoff value. The overall agreement of PCPI and HPLC-MS-MS results was 98.7%. The sensitivity of the PCPI was 0.982 and the specificity 0.987. Testing at 100 ng/mL of other drugs or their metabolites demonstrated no cross-reactivity. The within-run precision was CV = 2% (n = 12); the between-run precision was CV = < 6% (n = 4). The assay was found linear from -50% to 150% (12.5-37.5 ng/mL) of the cutoff concentration. The Lin-Zhi PCPI provides a precise, reliable method for the routine detection of phencyclidine in urine specimens.     

Method development and validation of almotriptan in human plasma by HPLC tandem mass spectrometry: application to pharmacokinetic study

A simple, sensitive and selective method has been developed for quantification of Almotriptan (AL) in human plasma using Almotriptan-d(6) (ALD6) as an internal standard. Almotriptan and Almotriptan-d(6) were detected with proton adducts at m/z 336.1→201.1 and 342.2→207.2 in multiple reaction monitoring (MRM) positive mode, respectively. The method was linear over a concentration range of 0.5-150.0 ng/mL. The limit of detection (LOD) and limit of quantification (LOQ) for Almotriptan were 0.2 pg/mL and 0.5 ng/mL, respectively. Liquid-liquid extraction was used followed by MS/MS (ion spray). The method was shown to be precise with an average within-run and between-run variation of 0.68 to 2.78% and 0.57 to 0.86%, respectively. The average within-run and between-run accuracy of the method throughout its linear range was 98.94 to 102.64% and 99.43 to 101.44%, respectively. The mean recovery of drug and internal standard from human plasma was 92.12 ± 4.32% and 89.62 ± 6.32%. It can be applied for clinical and pharmacokinetic studies.