改良ALT和AST参考方法的建立、评价及应用
|
摘要
1研究意义与目的
检验结果的互认是当前临床实验室质量管理中的研究热点之一。互认的基础是检验结果具有可比性,而检测结果只有直接或间接地溯源到参考方法和或参考物质,才能保证检验结果的准确性和不同实验室检测结果具有可比性。ALT和AST是临床实验室血清酶学测定中非常重要的常规项目,其测定的准确性直接影响到肝脏疾病的诊断、治疗和预后。目前国内临床实验室血清酶测定大多采用非配套检测系统,即大多采用进口仪器和校准品,但试剂为国产试剂,或改变配套检测系统的检验程序,致使校准品丧失溯源性,导致不同检测系统或方法的检验结果存在较大差异,缺乏可比性。而且国产ALT和AST试剂几乎全是不含磷酸吡哆醛的,而IFCC公布的37℃条件下ALT和AST的参考方法中是含有磷酸吡哆醛的。鉴于此,本研究将参照IFCC推荐的参考方法,建立不含磷酸吡哆醛的改良ALT和AST参考方法,并对初步建立的改良ALT和AST参考方法进行方法学性能评价,了解其各项性能指标。同时用建立的改良ALT和AST参考方法对制备的酶校准品进行赋值,然后用赋值的酶校准品校准各实验室开放系统,评价校准前后各系统与参考方法间测定结果的偏倚,同时了解不同实验室间ALT和AST测定结果的可比性以及非新鲜血清样本的基质效应。
2方法
参照IFCC公布的37℃条件下ALT和AST一级参考测量程序相关文件,建立本研究不含磷酸吡哆醛的改良ALT和AST参考方法。根据美国国家实验室标准化协会(CLSI)系列文件(EP5-A2、EP15-A、EP6-A)对建立的改良参考方法的精密度、正确度、线性范围等性能进行评价。使用初步建立的改良ALT和AST参考方法对新鲜血清进行定值,用此定值的新鲜血清作为校准品校准广州地区10家实验室的常规检测系统。用改良参考方法和常规检测系统同时检测7份能力验证盲样、4份市售制备物和5份新鲜血清,比较校准前后测定结果间的偏倚,比较常规方法与参考方法间基于不同基质(人血清与质控血清)的偏倚差异,进行检验结果的可比性和基质效应的评价。
3结果
紫外分光光度计、分析天平、pH计、电子温度计、移液器和容量瓶均检定合格,不确定度符合方法要求。温控器显示为37.2℃时,比色杯内溶液温度最接近37℃;比色杯内溶液180秒后可达到设定温度;移液器加样模式实验显示吸一档打两档,打时靠壁为精密度最佳的加样模式。改良ALT和AST参考方法的批内不精密度均<1%,总不精密度均<2%,符合精密度性能要求;检测日本临床化学会酶参考品结果均在其不确定度允许范围内,cfas各浓度测定结果与靶值基本一致,参加参考实验室网络赋值计划测定结果与均值比较在等效限内。改良ALT和AST参考方法分析测量范围上限分别为335.8U/L和363.6U/L。校准前ALT和AST新鲜血清测定结果与参考方法测定结果之间偏倚大于20%的所有11个测定结果中,有10个测定结果来源于用理论K值计算酶活性的检测系统;校准前新鲜血清组ALT和AST的最大偏倚分别为-29.2%和-25.0%,平均偏倚分别为10.7%和9.3%;使用改良参考方法定值的酶校准品校准后,最大偏倚分别降为15.2%和12.3%,平均偏倚分别降为3.4%和2.1%。ALT结果中1种能力验证盲样对所有检测系统都表现基质效应,AST结果中2种能力验证盲样对所有检测系统无基质效应,其他样品视系统的不同而表现出不同的情况。
4结论
改良ALT和AST参考方法已基本建立。精密度、准确度及线性范围等性能指标均符合方法要求。应用本改良参考方法定值的新鲜血清作为校准品进行校准是实现不同检测系统检验结果一致性和可比性的有效途径,样品的基质效应影响血清ALT和AST测定的准确性和可比性,应充分重视基质效应对临床检验量值溯源的影响
1 Objective
Nowadays,“mutual Recognition”of clinical results is the hot spot of quality management in Clinical Laboratory. The precondition of mutual recognition is comparability of clinical results. Only when the reference method is directly or indirectly traceable, can the accuracy and comparability among different laboratories be assured. ALT and AST are very important routine tests in serum enzymes determination. Diagnosis, treatment and prognosis of liver diseases are subject to the accuracy of the test. At present, domestic clinical laboratory usually adopt non-matching analytical system, i.e., using import instrument and calibrator but domestic reagent, or changing the test procedure test system. These practices destroy the traceability of calibrator. The results of different test system or method are significantly different and incomparable. In addition, domestic reagent of ALT and AST are almost phosphopyridoxal free, while IFCC reference method for ALT and AST at 37℃contains phosphopyridoxal. In consideration of it, this research shall base on the reference method recommended by IFCC, to find out the ameliorative reference method for phosphopyridoxal free ALT and AST, and carry out methodological evaluation on this preliminary ameliorative reference method to study its performance. Meanwhile, a enzyme calibrator was prepared and its value for ALT and AST was assigned with this preliminary ameliorative ALT and AST reference method, and find out the bias of test results of open and limited system among different laboratories as well as different reference methods, so as to measure the comparability and matrix effect of ALT and AST results and among different laboratories.
2. Methods
This ameliorative reference method for phosphopyridoxal free ALT and AST is based on the IFCC primary reference measurement procedure of ALT and AST at 37℃. The precision, accuracy, linearity of the method is measured as per CLSI protocols (EP5-A2, EP15-A, EP6-A). The ameliorative reference method is used to determine fresh serum, which will be the calibrator for the routine analytical systems in the 10 laboratories in Guangzhou. The activity of external quality assessment materials、products produced by factories、fresh sera were measured by the routine analytical systems and the ameliorative reference procedure. The comparison is to find out bias before and after calibration between routine method and ameliorative reference methods, bias between routine method and ameliorative reference method for different medium (human serum and quality control serum). Comparability and matrix effect evaluated this way.
3. Result
Ultraviolet spectrophotometer, analytic balance, pH meter, electronic thermometer, pipettes and volumetirc flask have been verified and its uncertainty fit to method’s requirements. When the temperature controller showed 37.2℃, the temperature in cuvette was equal to 37.0℃. The temperature in cuvette reaches the set temperature after 180 seconds. In transferpettor sample input experiment show switch to dimension 2 when the absorbance measurement indicate 1 and inputting sample by side will generate the best precision. The imprecision within run of the ameliorative reference method is less than 1%, total imprecision less than 2%. Both satisfy precision requirement.The results of JSCC ERM which determine by the ameliorative reference method were within uncertainty tolerance. The results of cfas conformed to target. Bias between involved laboratories’network evaluation result and average is within equivalent area. The upper limit of the measurement range of ALT and AST was 335.8U/L and 363.6U/L respectively. Among the 11 tests, of which the bias of ALT and AST fresh serum before calibration and reference method is over 20%, 10 were enzyme activity computed with theoretical K value. The maximum bias of ALT and AST of fresh serum before calibration was -29.2% and -25.0% respectively, average at 10.7% and 9.3%. Calibrated with enzyme calibrator of the ameliorative reference method, the maximum bias decreases to 15.2% and 12.3%, average at 3.4% and 2.1%. In ALT test result, One external quality assessment materials showed matrix effect on all the routine analytical systems. In AST test result, two external quality assessment materials showed no matrix effect on all the routine analytical systems. The routine analytical systems dependent matrix effects of other materials were observed on other materials.
4. Conclusion
Ameliorative reference method has been established basically. Precision, accuracy and linear range and other performance all satisfy the requirements. It’s an effective way to achieve traceability in different testing systems by using the determined fresh serum as the calibrator. Matrix effect will affect the accuracy and comparability of ALT and AST test. It’s important to attach importance to the influence of matrix effect on clinical result traceability.
检验结果的互认是当前临床实验室质量管理中的研究热点之一。互认的基础是检验结果具有可比性,而检测结果只有直接或间接地溯源到参考方法和或参考物质,才能保证检验结果的准确性和不同实验室检测结果具有可比性。ALT和AST是临床实验室血清酶学测定中非常重要的常规项目,其测定的准确性直接影响到肝脏疾病的诊断、治疗和预后。目前国内临床实验室血清酶测定大多采用非配套检测系统,即大多采用进口仪器和校准品,但试剂为国产试剂,或改变配套检测系统的检验程序,致使校准品丧失溯源性,导致不同检测系统或方法的检验结果存在较大差异,缺乏可比性。而且国产ALT和AST试剂几乎全是不含磷酸吡哆醛的,而IFCC公布的37℃条件下ALT和AST的参考方法中是含有磷酸吡哆醛的。鉴于此,本研究将参照IFCC推荐的参考方法,建立不含磷酸吡哆醛的改良ALT和AST参考方法,并对初步建立的改良ALT和AST参考方法进行方法学性能评价,了解其各项性能指标。同时用建立的改良ALT和AST参考方法对制备的酶校准品进行赋值,然后用赋值的酶校准品校准各实验室开放系统,评价校准前后各系统与参考方法间测定结果的偏倚,同时了解不同实验室间ALT和AST测定结果的可比性以及非新鲜血清样本的基质效应。
2方法
参照IFCC公布的37℃条件下ALT和AST一级参考测量程序相关文件,建立本研究不含磷酸吡哆醛的改良ALT和AST参考方法。根据美国国家实验室标准化协会(CLSI)系列文件(EP5-A2、EP15-A、EP6-A)对建立的改良参考方法的精密度、正确度、线性范围等性能进行评价。使用初步建立的改良ALT和AST参考方法对新鲜血清进行定值,用此定值的新鲜血清作为校准品校准广州地区10家实验室的常规检测系统。用改良参考方法和常规检测系统同时检测7份能力验证盲样、4份市售制备物和5份新鲜血清,比较校准前后测定结果间的偏倚,比较常规方法与参考方法间基于不同基质(人血清与质控血清)的偏倚差异,进行检验结果的可比性和基质效应的评价。
3结果
紫外分光光度计、分析天平、pH计、电子温度计、移液器和容量瓶均检定合格,不确定度符合方法要求。温控器显示为37.2℃时,比色杯内溶液温度最接近37℃;比色杯内溶液180秒后可达到设定温度;移液器加样模式实验显示吸一档打两档,打时靠壁为精密度最佳的加样模式。改良ALT和AST参考方法的批内不精密度均<1%,总不精密度均<2%,符合精密度性能要求;检测日本临床化学会酶参考品结果均在其不确定度允许范围内,cfas各浓度测定结果与靶值基本一致,参加参考实验室网络赋值计划测定结果与均值比较在等效限内。改良ALT和AST参考方法分析测量范围上限分别为335.8U/L和363.6U/L。校准前ALT和AST新鲜血清测定结果与参考方法测定结果之间偏倚大于20%的所有11个测定结果中,有10个测定结果来源于用理论K值计算酶活性的检测系统;校准前新鲜血清组ALT和AST的最大偏倚分别为-29.2%和-25.0%,平均偏倚分别为10.7%和9.3%;使用改良参考方法定值的酶校准品校准后,最大偏倚分别降为15.2%和12.3%,平均偏倚分别降为3.4%和2.1%。ALT结果中1种能力验证盲样对所有检测系统都表现基质效应,AST结果中2种能力验证盲样对所有检测系统无基质效应,其他样品视系统的不同而表现出不同的情况。
4结论
改良ALT和AST参考方法已基本建立。精密度、准确度及线性范围等性能指标均符合方法要求。应用本改良参考方法定值的新鲜血清作为校准品进行校准是实现不同检测系统检验结果一致性和可比性的有效途径,样品的基质效应影响血清ALT和AST测定的准确性和可比性,应充分重视基质效应对临床检验量值溯源的影响
1 Objective
Nowadays,“mutual Recognition”of clinical results is the hot spot of quality management in Clinical Laboratory. The precondition of mutual recognition is comparability of clinical results. Only when the reference method is directly or indirectly traceable, can the accuracy and comparability among different laboratories be assured. ALT and AST are very important routine tests in serum enzymes determination. Diagnosis, treatment and prognosis of liver diseases are subject to the accuracy of the test. At present, domestic clinical laboratory usually adopt non-matching analytical system, i.e., using import instrument and calibrator but domestic reagent, or changing the test procedure test system. These practices destroy the traceability of calibrator. The results of different test system or method are significantly different and incomparable. In addition, domestic reagent of ALT and AST are almost phosphopyridoxal free, while IFCC reference method for ALT and AST at 37℃contains phosphopyridoxal. In consideration of it, this research shall base on the reference method recommended by IFCC, to find out the ameliorative reference method for phosphopyridoxal free ALT and AST, and carry out methodological evaluation on this preliminary ameliorative reference method to study its performance. Meanwhile, a enzyme calibrator was prepared and its value for ALT and AST was assigned with this preliminary ameliorative ALT and AST reference method, and find out the bias of test results of open and limited system among different laboratories as well as different reference methods, so as to measure the comparability and matrix effect of ALT and AST results and among different laboratories.
2. Methods
This ameliorative reference method for phosphopyridoxal free ALT and AST is based on the IFCC primary reference measurement procedure of ALT and AST at 37℃. The precision, accuracy, linearity of the method is measured as per CLSI protocols (EP5-A2, EP15-A, EP6-A). The ameliorative reference method is used to determine fresh serum, which will be the calibrator for the routine analytical systems in the 10 laboratories in Guangzhou. The activity of external quality assessment materials、products produced by factories、fresh sera were measured by the routine analytical systems and the ameliorative reference procedure. The comparison is to find out bias before and after calibration between routine method and ameliorative reference methods, bias between routine method and ameliorative reference method for different medium (human serum and quality control serum). Comparability and matrix effect evaluated this way.
3. Result
Ultraviolet spectrophotometer, analytic balance, pH meter, electronic thermometer, pipettes and volumetirc flask have been verified and its uncertainty fit to method’s requirements. When the temperature controller showed 37.2℃, the temperature in cuvette was equal to 37.0℃. The temperature in cuvette reaches the set temperature after 180 seconds. In transferpettor sample input experiment show switch to dimension 2 when the absorbance measurement indicate 1 and inputting sample by side will generate the best precision. The imprecision within run of the ameliorative reference method is less than 1%, total imprecision less than 2%. Both satisfy precision requirement.The results of JSCC ERM which determine by the ameliorative reference method were within uncertainty tolerance. The results of cfas conformed to target. Bias between involved laboratories’network evaluation result and average is within equivalent area. The upper limit of the measurement range of ALT and AST was 335.8U/L and 363.6U/L respectively. Among the 11 tests, of which the bias of ALT and AST fresh serum before calibration and reference method is over 20%, 10 were enzyme activity computed with theoretical K value. The maximum bias of ALT and AST of fresh serum before calibration was -29.2% and -25.0% respectively, average at 10.7% and 9.3%. Calibrated with enzyme calibrator of the ameliorative reference method, the maximum bias decreases to 15.2% and 12.3%, average at 3.4% and 2.1%. In ALT test result, One external quality assessment materials showed matrix effect on all the routine analytical systems. In AST test result, two external quality assessment materials showed no matrix effect on all the routine analytical systems. The routine analytical systems dependent matrix effects of other materials were observed on other materials.
4. Conclusion
Ameliorative reference method has been established basically. Precision, accuracy and linear range and other performance all satisfy the requirements. It’s an effective way to achieve traceability in different testing systems by using the determined fresh serum as the calibrator. Matrix effect will affect the accuracy and comparability of ALT and AST test. It’s important to attach importance to the influence of matrix effect on clinical result traceability.
引文
[1]Panteghinia M,Forestb J C.Standardization in laboratory medicine:New challenges[J].Clinica Chimica Acta,2005,355(1):1.12.
[2]Thiempont L M,Vytfanghe K V,Leenheer A P,et a1.Reference measurement system in clinical chemistry[J].Clinca Chimica Acta,2002,323(1):73-87.
[3]Klaus Lorentz.IFCC primary reference procedures for the measuremeat of catalytic concentrations of enzymes-the new concept[J].Clin Chem Lab Med,2002,40(6):549.
[4]Siekmann L,Bonora R,Ceriotti F,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃.Part 1.The concept of reference procedures for the measurement of catalytic activity concentrations of enzymes[J].Clin Chem Lab Med,2002,40(7):631-634.
[5]Schumann g,Bonora R,Ceriotti F,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃.Part 2.Reference procedures for the measurement of catalytic activity concentrations of creatine kinase[J].Clin Chem Lab Med,2002,40(7):635-642.
[6]Schumann g,Bonora R,Ceriotti F,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃. Part 3.Reference procedures for the measurement of catalytic activity concentrations of lactate dehydrogenase[J].Clin Chem Lab Med,2002, 40(7):643-648.
[7]Schumann g,Bonora R,Ceriotti F,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃.Part 4.Reference procedures for the measurement of catalytic activity concentrations of alanine aminotransferase[J].Clin Chem Lab Med,2002, 40(7):718-724.
[8]Schumann g,Bonora R,Ceriotti F,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃.Part5.Reference procedures for the measurement of catalytic activity concentrations of aspartate aminotransferase[J].Clin Chem Lab Med,2002,40(7):725-733.
[9]Schumann g,Bonora R,Ceriotti F,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃.Part 6.Reference procedures for the measurement of catalytic activity concentrations of gamma-glutamyltransferase[J].Clin Chem Lab Med,2002,40(7):734-738.
[10] Zhou SL, Gordon RE, BradburyM, et al. Ethanol up-regulates fatty acid up take and plasma membrane expression and export of mitochondrial aspartate aminotransferase in HepG2 cells[J]. Hepatology,1998,27:1064-1074.
[11]梁淑慧,陈小红,肖木洲,等.肝硬化患者ASTALT比值的改变及其临床意义的探讨[J].广西医学,2003,25(12):2383-2384.
[12]张秀明,庄俊华,徐宁,等.不同检测系统4种心肌酶测定结果的比对与临床可接受性评价[J].临床检验杂志,2005,23(6):404-407.
[13]张秀明,庄俊华,徐宁,等.不同检测系统血清酶测定结果的偏倚评估与可比性研究[J].中华检验医学杂志,2006,29(4):346-349.
[14]张秀明,李炜煊,郑松柏,等.不同检测系统17项常规生化结果的比对和偏倚评估[J].检验医学,2007,22(2):164-168.
[15] NCCLS. EP5-A2 Evaluation of precision performance of quantative measurement methods;approved guideline-second edition.Wayne,PA:NCCLS,2004.
[16] NCCLS. EP15-A User demonstration of performance for precision and accuracy;approved guideline. Wayne,PA:NCCLS,2001.
[17] NCCLS. EP6-A Evaluation of the linearity of quantitative measurement procedures:a statistical approach; approved guideline. Wayne,PA:NCCLS,2003.
[18]毕波,吕元.定量检测方法学性能验证的系统设计[J].中华检验医学杂志2007,30(2): 143-145.
[19] Stockl D,Franzini C,Kratochvila J,et al.Analytical specifications of reference methods compilation and critical discussion[J]. Clin Chem ClinBiochem,1996,34:319-337.
[20] Thienpont L, Franzini C,Kratochvila J,et al.Analytical specifications of reference methods and operating specifications for networks of reference laboratories[J]. Clin Chem Clin Biochem,1995,33:3949-957.
[21]Panteghinia M,Ceriotti F,Schumann G, et al.Establishing a reference system in clinical enzymology[J].Clin Chem Lab Med,2001;39:795-800.
[22]Panteghini M, Pagani F. AACC creatine kinase MB (CKMB) standardization material used as manufacturer’s working calibrator is unable to harmonize CK-MB results between two commercial immunoassays[J]. Clin Chem. 2004; 50:1711-2.
[23]Medical laboratories-Particular requirements for quality and competence[S].International Standard ISO 15189 2002:3.15
[24]In vitro diagnostic medical devices-Measurement of quantities in biological samples-Metrological traceability of values assigned to calibrators and control materials [S]. International Standard ISO 17511 2003,4.2.2-4.2.4.
[25]Muller MM: Reference materials: general aspects and IFCC straegy [J]. Clin Biochem,1998;31:433-436.
[26]Dybkae.The tofluous road to the adoption of katal for the expression of catalytic Activity by the general conference on weights and measures [J].Clinical Chemistry,2002,48(3):586-590.
[27]Bowers JR, Bergmeyer HU, Morder M, et al. Approved recommendations on IFCC methods for the measurements of catalytic concentrations of enzymes. part 1. General considerations concerning the blood serum or plasma of man. Clin Chim Acta, 1979, 98:163F-174F.
[28]Shaw LM, Stromme JH, London JL, et al. IFCC method forγ-glutamyltrans- ferase[J]. Clin Chem Clin Biochem, 1983, 21:633-646.
[29]Bergmeyer HU, Horder M, Rej R. IFCC methods for alanine aminotrans- ferase[J].Clin Chem Clin Biochem, 1986, 24:481-495.
[30]Bergmeyer HU, Horder M, Rej R. IFCC methods for aspartate aminotrans- ferase[J]. Clin Chem Clin Biochem, 1986, 24:497-510.
[31]杨振华.中华医学会检验学会酶活性浓度测定的推荐方法,测定人血清(血浆)中酶活性浓度方法总则(草案) [J].中华医学检验杂志,1994,2:121-123.
[32]Schumann g,Aoki R,Ferrero CA,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃.Part 2.Reference procedures for the measurement of catalytic activity concentrations of a-amylase[J].Clin Chem Lab Med,2006,44(9):1146-1155.
[33]陈文祥,申子瑜,杨振华,等.临床生化检验参考方法及其主要分析原理[J].中华检验医学杂志,2007,30(3):245-248.
[34] Siekmann L,Bonora R,Durtis CA,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃.Part 1.Certification of reference materials for the determination of enzymatic activity concentrations of gamma-glutamyltransferase, lactate dehydrogenase, alanine aminotransferase and creatine kinase accord[J].Clin Chem Lab Med,2002,40(7):739-745.
[35]Canalias F, CamprubíS, Sánchez M, et a1.Metrological traceability of values for catalytic concentration of enzymes assigned to a calibration material[J]. Clin Chem Lab Med. 2006;44(3):333-339.
[36]Georges F,Francoise I B,Djamila M,et a1.A reference material for traceability of aspartate aminotransferase(AST)results[J].Clin Chem Lab Med,2005,43(5):549-553.
[37]Siekman L,Doumas BT,Thienpon L,et al. Reference materials and reference measurement systems in laboratory medicine. Network of reference laboratories[J]. Eur J Clin Chem Clin Biochem.1995,33:1013-1017.
[38]汪静,张传宝,居漪,等.国际临床化学联合会酶学测定参考方法实验室网络的初步建立[J].中华检验医学杂志,2008,31(3):258-263.
[39]王涛,齐丽丽,刘海燕,等.六种临床诊断酶学测定参考方法的应用及国际比对结果分析[J].中华检验医学杂志,2008,31(3):264-269.
[40]赵海舰,张天娇,张传宝,等.血清尿素测定基质效应研究[J].中华检验医学杂志,2008,31(3):270-275.
[41]王丽,权翠侠,牛璐璐,等.新鲜血清赋值传递提高不同生化检测系统检验结果的可比性[J].临床检验杂志,2008,26(4):294-296.
[42] Rodríguez-Cabaleiro D,Van Uytfanghe K, Stove V,et al. Pilot study for the standardization of insulin immunoassays with isotope dilution liquid chromatography/tandem mass spectrometry[J]. Clin Chem. 2007;53(8): 1462-1469.
[43]彭黎明,王伟鑫,聂李平,等.用参考方法验证多台血液分析仪的校准结果[J].临床检验杂志,2007,25(6):463-465.
[44]董军,李红霞,满永,等.血清总胆固醇、总甘油、游离甘油和甘油三脂标准物质的研制[J].中华检验医学杂志,2008,31(3):276-279.
[45]李淑葵,何振坤,徐国宾,等.血清γ-谷氨酰转移酶测定参考方法的建立及在酶校准品定值中的应用[J].中华医学检验杂志,2007,30(6):679-682.
[2]Thiempont L M,Vytfanghe K V,Leenheer A P,et a1.Reference measurement system in clinical chemistry[J].Clinca Chimica Acta,2002,323(1):73-87.
[3]Klaus Lorentz.IFCC primary reference procedures for the measuremeat of catalytic concentrations of enzymes-the new concept[J].Clin Chem Lab Med,2002,40(6):549.
[4]Siekmann L,Bonora R,Ceriotti F,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃.Part 1.The concept of reference procedures for the measurement of catalytic activity concentrations of enzymes[J].Clin Chem Lab Med,2002,40(7):631-634.
[5]Schumann g,Bonora R,Ceriotti F,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃.Part 2.Reference procedures for the measurement of catalytic activity concentrations of creatine kinase[J].Clin Chem Lab Med,2002,40(7):635-642.
[6]Schumann g,Bonora R,Ceriotti F,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃. Part 3.Reference procedures for the measurement of catalytic activity concentrations of lactate dehydrogenase[J].Clin Chem Lab Med,2002, 40(7):643-648.
[7]Schumann g,Bonora R,Ceriotti F,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃.Part 4.Reference procedures for the measurement of catalytic activity concentrations of alanine aminotransferase[J].Clin Chem Lab Med,2002, 40(7):718-724.
[8]Schumann g,Bonora R,Ceriotti F,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃.Part5.Reference procedures for the measurement of catalytic activity concentrations of aspartate aminotransferase[J].Clin Chem Lab Med,2002,40(7):725-733.
[9]Schumann g,Bonora R,Ceriotti F,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃.Part 6.Reference procedures for the measurement of catalytic activity concentrations of gamma-glutamyltransferase[J].Clin Chem Lab Med,2002,40(7):734-738.
[10] Zhou SL, Gordon RE, BradburyM, et al. Ethanol up-regulates fatty acid up take and plasma membrane expression and export of mitochondrial aspartate aminotransferase in HepG2 cells[J]. Hepatology,1998,27:1064-1074.
[11]梁淑慧,陈小红,肖木洲,等.肝硬化患者ASTALT比值的改变及其临床意义的探讨[J].广西医学,2003,25(12):2383-2384.
[12]张秀明,庄俊华,徐宁,等.不同检测系统4种心肌酶测定结果的比对与临床可接受性评价[J].临床检验杂志,2005,23(6):404-407.
[13]张秀明,庄俊华,徐宁,等.不同检测系统血清酶测定结果的偏倚评估与可比性研究[J].中华检验医学杂志,2006,29(4):346-349.
[14]张秀明,李炜煊,郑松柏,等.不同检测系统17项常规生化结果的比对和偏倚评估[J].检验医学,2007,22(2):164-168.
[15] NCCLS. EP5-A2 Evaluation of precision performance of quantative measurement methods;approved guideline-second edition.Wayne,PA:NCCLS,2004.
[16] NCCLS. EP15-A User demonstration of performance for precision and accuracy;approved guideline. Wayne,PA:NCCLS,2001.
[17] NCCLS. EP6-A Evaluation of the linearity of quantitative measurement procedures:a statistical approach; approved guideline. Wayne,PA:NCCLS,2003.
[18]毕波,吕元.定量检测方法学性能验证的系统设计[J].中华检验医学杂志2007,30(2): 143-145.
[19] Stockl D,Franzini C,Kratochvila J,et al.Analytical specifications of reference methods compilation and critical discussion[J]. Clin Chem ClinBiochem,1996,34:319-337.
[20] Thienpont L, Franzini C,Kratochvila J,et al.Analytical specifications of reference methods and operating specifications for networks of reference laboratories[J]. Clin Chem Clin Biochem,1995,33:3949-957.
[21]Panteghinia M,Ceriotti F,Schumann G, et al.Establishing a reference system in clinical enzymology[J].Clin Chem Lab Med,2001;39:795-800.
[22]Panteghini M, Pagani F. AACC creatine kinase MB (CKMB) standardization material used as manufacturer’s working calibrator is unable to harmonize CK-MB results between two commercial immunoassays[J]. Clin Chem. 2004; 50:1711-2.
[23]Medical laboratories-Particular requirements for quality and competence[S].International Standard ISO 15189 2002:3.15
[24]In vitro diagnostic medical devices-Measurement of quantities in biological samples-Metrological traceability of values assigned to calibrators and control materials [S]. International Standard ISO 17511 2003,4.2.2-4.2.4.
[25]Muller MM: Reference materials: general aspects and IFCC straegy [J]. Clin Biochem,1998;31:433-436.
[26]Dybkae.The tofluous road to the adoption of katal for the expression of catalytic Activity by the general conference on weights and measures [J].Clinical Chemistry,2002,48(3):586-590.
[27]Bowers JR, Bergmeyer HU, Morder M, et al. Approved recommendations on IFCC methods for the measurements of catalytic concentrations of enzymes. part 1. General considerations concerning the blood serum or plasma of man. Clin Chim Acta, 1979, 98:163F-174F.
[28]Shaw LM, Stromme JH, London JL, et al. IFCC method forγ-glutamyltrans- ferase[J]. Clin Chem Clin Biochem, 1983, 21:633-646.
[29]Bergmeyer HU, Horder M, Rej R. IFCC methods for alanine aminotrans- ferase[J].Clin Chem Clin Biochem, 1986, 24:481-495.
[30]Bergmeyer HU, Horder M, Rej R. IFCC methods for aspartate aminotrans- ferase[J]. Clin Chem Clin Biochem, 1986, 24:497-510.
[31]杨振华.中华医学会检验学会酶活性浓度测定的推荐方法,测定人血清(血浆)中酶活性浓度方法总则(草案) [J].中华医学检验杂志,1994,2:121-123.
[32]Schumann g,Aoki R,Ferrero CA,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃.Part 2.Reference procedures for the measurement of catalytic activity concentrations of a-amylase[J].Clin Chem Lab Med,2006,44(9):1146-1155.
[33]陈文祥,申子瑜,杨振华,等.临床生化检验参考方法及其主要分析原理[J].中华检验医学杂志,2007,30(3):245-248.
[34] Siekmann L,Bonora R,Durtis CA,et a1.IFCC primary reference procedures for the measurement of catalytic activity concentrations of enzymes at 37℃.Part 1.Certification of reference materials for the determination of enzymatic activity concentrations of gamma-glutamyltransferase, lactate dehydrogenase, alanine aminotransferase and creatine kinase accord[J].Clin Chem Lab Med,2002,40(7):739-745.
[35]Canalias F, CamprubíS, Sánchez M, et a1.Metrological traceability of values for catalytic concentration of enzymes assigned to a calibration material[J]. Clin Chem Lab Med. 2006;44(3):333-339.
[36]Georges F,Francoise I B,Djamila M,et a1.A reference material for traceability of aspartate aminotransferase(AST)results[J].Clin Chem Lab Med,2005,43(5):549-553.
[37]Siekman L,Doumas BT,Thienpon L,et al. Reference materials and reference measurement systems in laboratory medicine. Network of reference laboratories[J]. Eur J Clin Chem Clin Biochem.1995,33:1013-1017.
[38]汪静,张传宝,居漪,等.国际临床化学联合会酶学测定参考方法实验室网络的初步建立[J].中华检验医学杂志,2008,31(3):258-263.
[39]王涛,齐丽丽,刘海燕,等.六种临床诊断酶学测定参考方法的应用及国际比对结果分析[J].中华检验医学杂志,2008,31(3):264-269.
[40]赵海舰,张天娇,张传宝,等.血清尿素测定基质效应研究[J].中华检验医学杂志,2008,31(3):270-275.
[41]王丽,权翠侠,牛璐璐,等.新鲜血清赋值传递提高不同生化检测系统检验结果的可比性[J].临床检验杂志,2008,26(4):294-296.
[42] Rodríguez-Cabaleiro D,Van Uytfanghe K, Stove V,et al. Pilot study for the standardization of insulin immunoassays with isotope dilution liquid chromatography/tandem mass spectrometry[J]. Clin Chem. 2007;53(8): 1462-1469.
[43]彭黎明,王伟鑫,聂李平,等.用参考方法验证多台血液分析仪的校准结果[J].临床检验杂志,2007,25(6):463-465.
[44]董军,李红霞,满永,等.血清总胆固醇、总甘油、游离甘油和甘油三脂标准物质的研制[J].中华检验医学杂志,2008,31(3):276-279.
[45]李淑葵,何振坤,徐国宾,等.血清γ-谷氨酰转移酶测定参考方法的建立及在酶校准品定值中的应用[J].中华医学检验杂志,2007,30(6):679-682.