四种国产和两种进口酶法肌酐试剂的分析性能及准确度验证

摘要

Objective To assessed the analytical performance of six commercial enzymatic creatinine (Cr) reagents and compared the accuracy among them.Four reagents came from domestic manufactures (Maker Biotechnology,Beijing Strong Biotechnologies,Biosino Bio-technology & Science,BaiDing Biological Engineering,labeled as A,B,C,D respectively) and the other two reagents came from oversea manufactures (Sekisui Medical Technology,Japan and Wako Pure Chemical Industries,Japan,labeled as E,F respectively).Methods Performance verification test.70 serum samples were obtained from chronic kidney disease outpatients and medical center healthy volunteers in Peking Union Medical College Hospital from May 2010.Among 30 patients ureh CKD (15 moles,15 females)aged 18 to 80 years old;40 healthy volunteers (20 males,20 females) aged 20 to 60 years old.To evaluate the within-run imprecision and total imprecision according to the CLSI EP15-A protocol,two level control materials were measured with six reagents in five days,with a frequency of four times a day.To verify the linear ranges according to the EP6-A protocol,a series of samples that mixed by high and low level serum or urine pools with specific ratio were tested.To evaluate the anti-interference ability,we tested the sera added specific concentration of substances (bilirubin,hemoglobin,chyle).To verify the reasonable dilution ratio,diluted sera in different ratio were tested.Correlations and deviation among the detection results of six reagents were evaluated by testing 50 fresh patient samples.Accuracy was evaluated by measuring the NIST SRM 967a and NIST SRM 909b standard substances.Results There were no significant differences between the domestic and imported Cr reagents,with the within-run CV ranging from 0.5％ to 1.2％ and the total CV ranging from 0.5％ to 1.9％,all of which were lower than the allowed biological variation for imprecision (3.0％).Linear ranges of the six Cr reagents were all relatively wide.No significant interference (bias％ ≤ ± 10％)was found in the test of six reagents when conjugated bilirubin ≤ 332 μmol/L,unconjugated bilirubin ≤ 327μmol/L,or chyle turbidity ≤ 1530 FTU; no significant interference was found for the test of C,D,F reagents when Hb≤4.83 g/L; and for A,B,E when Hb ≤3.381,3.864,4.347g/L,respectively.Percent deviation of the measured results of dulited serum samples diluted by 2 to 16 times compared to the results of the original samples ranged from 95.96％ to 108.62％.Fresh patient samples comparison tests exhibited good correlation among the six reagents (r =1.000),although there were some differences (-4.91％ to-4.75％).For NIST 967a,the percent bias ranged from-1.85％ to 2.81％ (level 1,66.5 μmol/L),and from-2.28％ to 2.05％ (level 2,346.2 μmol/L),none of the six reagents exceeded the allowed Cr bias derived from biological variation (4.0％).For NIST 909b,the percent bias ranged from-13.4％ to 1.01％ for level 1 (56.18 μmol/L),while its value ranged from-6.49％ to -1.79％ for level 2 (467.4 μmol/L),and these biases all exceeded the allowable range of target value.Conclusions Assessment of four domestic enzymatic Cr reagents and two imported reagents show high precision,wide linear range,and their dilution accuracy can meet the clinical needs.Moreover,the six regents show good anti-interference capability (bilirubin,hemoglobin,chyle) and high accuracy.No significant differences have been found in the above assessed properties among these reagents.%目的 比较4种市售国产酶法肌酐(Cr)(迈克、九强、中生、柏定,标为A、B、C、D)和2种进口酶法Cr试剂(积水、和光,标为E、F)的分析性能及准确度差异.方法 实验性能验证.从2010年5月就诊于我院肾内科的患者和体检中心查体人群中选择慢性肾脏病患者和表面健康人的血清标本共70例进行性能验证实验.其中CKD患者30例(男15例,女15例),年龄18 ～ 80岁.表面健康人40名(男20名,女20名),年龄20 ～ 60岁.参考CLSI EP15-A方案用6种试剂分别测定2个水平质控血清,每天测4次,连续测5d,计算批内及总不精密度(CV)；参考EP6-A方案,测定不同比例的高、低浓度混合血清和混合尿液标本评价线性范围；部分参考EP7-P方案,用测定含特定浓度干扰物(胆红素、血红蛋白、乳糜)的血清以评价抗干扰能力；验证血清样本的稀释准确性；测定50份新鲜患者血清比较各试剂测定结果的相关性和偏差.计算各试剂测定NIST SRM 967a、NIST SRM 909b标准物质的偏倚以验证各试剂的准确度.结果 6种Cr试剂的批内CV在0.5％ ～1.2％之间,总CV在0.5％～1.9％之间,4种国产试剂与2种进口试剂的不精密度无明显差异,均小于来源于生物变异的允许不精密度(3.0％).4种国产与2种进口酶法Cr验证的线性范围均较宽.当血清中结合胆红素≤332 μmol/L、游离胆红素≤327 μmol/L、乳糜浊度≤1530 FTU时对6种Cr的测定均未产生明显干扰(百分偏差≤±10％)；血红蛋白(Hb)≤4.83 g/L时对试剂C、D、F检测Cr不受干扰；试剂A、B、E分别在Hb 3.381、3.864、4.347 g/L以下不受干扰.血清样本2～16倍稀释后与原倍测定结果的百分偏差在95.96％ ～ 108.62％之间.各试剂测定新鲜血血清样本结果与6种试剂测定均值之间的平均百分偏差在-4.91％ ～4.75％,相关均良好(r均为1.000).各试剂检测NIST 967a水平1(66.5 μmol/L)和水平2(346.2 μmol/L)的百分偏倚分别为-1.85％ ～2.81％和-2.28％～2.05％,均未超出Cr来源于生物变异的允许偏倚(4.0％).检测NIST 909b水平1(56.18 μmol/L)和水平2(467.4 μmol/L)的百分偏倚分别为-13.4％ ～ 1.01％和-6.49％～-1.79％,均超出靶值允许范围.结论 4种国产酶法Cr试剂和2种进口试剂精密度良好,线性范围宽,稀释准确性满足临床需要,抗干扰能力较强,准确度均较高,国产和进口试剂的以上性能未发现明显差异.