摘要目的 对3种应用于自动生化分析仪的颗粒增强免疫透射比浊法(PETIA)检测血Cys C试剂进行分析性能验证,并初步应用于临床.方法 在OlympusAU2700自动生化分析仪上对上海景源、北京利德曼和北京九强公司生产的颗粒增强免疫透射比浊法(PETIA)Cys C试剂(标为A、B、C)进行性能验证,参考CLSI EP6-A、EP15-A、EP7-P方案对3种方法的精密度、线性范围、干扰因素(胆红素、血红蛋白、乳糜)进行评估,并与德灵颗粒增强免疫散射比浊法(PENIA)试剂进行比对.参考CLSI C28-A2方案应用120名表观健康人血清对试剂A、B、C的参考范围进行验证,应用80例门诊患者血清检测结果评估试剂A、B、C与德灵PENIA 法Cys C试剂判定Cys C异常的一致率.结果 试剂A、B、C的批内CV分别为3.08%～3.20%、2.30%～4.15%和1.38%～1.53%,总CV分别为3.29%～3.44%、2.65%～5.18%和1.67%～1.69%,均小于试剂盒声明不精密度;试剂A、B、C的定量测定下限可分别达0.41、0.23、0.07 mg/L,基本满足检测要求;线性范围分别为0.22～7.26 mg/L (r=0.996),0.20～7.72 mg/L (r=0.999),0.20～7.62 mg/L (r=0.997),线性相关均良好.试验浓度内的3种干扰物(胆红素≤684 μmol/L、Hb≤9.7 g/L及乳糜浊度≤6 200 FTU)对试剂C无显著干扰(＜±10%);胆红素≤684 μmol/L、Hb≤6.79 g/L及乳糜浊度≤6 200 FTU时,对试剂B无显著干扰;胆红素≤684 μmol/L、Hb≤4.85 g/L及乳糜浊度≤1 240 FTU时,对试剂A无显著干扰.对患者乳糜血清标本采用高速离心后,试剂A、B、C测定Cys C浓度较离心前的平均百分偏倚分别为-8.31%、1.52%、1.32%.试剂A、B、C结果与德灵PENIA法试剂的回归方程分别为Y=0.787X+0.492(R2=0.976)、Y=1.098X+0.137(R2=0.982)、Y=1.037X+0.249(R2=0.996).试剂A、B、C与德灵PENIA 法Cys C试剂判定Cys C异常的一致率分别为80%(Kappa=0.615,P=0.000)、100%(Kappa=1.000,P=0.000)、91.2%(Kappa=0.824,P=0.000);改用临床初步评估的参考范围后,试剂A判定符合率可提高至98.8%(Kappa=0.974,P=0.000).结论 应用于自动生化分析仪的3种颗粒增强免疫透射比浊法测定Cys C试剂盒,具有较高的精密度和灵敏度,与德灵颗粒增强免疫散射比浊法测定相关良好;可满足临床测试要求.但抗干扰能力存在差异,部分试剂参考范围也有待进一步验证.

abstractsObjective To validate the analytical performance of three Cys C reagents with particle-enhanced turbidimetric immunoassay(PETIA) method used on the automatic biochemistry analyzer for preliminary clinical application.Methods The performance of three Cys C reagents (labeled as A, B, C) with PETIA method from Shanghai Jing Yuan Co., Beijing Leadman Co. and Beijing Jiuqiang Co. on OlympusAU2700 automatic biochemistry analyzer were assessed.According to the standard of CLSI EP6-A, EP15-A and EP7-P, the precision, linearity range, disturbance (bilirubin, hemoglobin, chyle) were assessed, and compared with those of Cys C reagent based on particle-enhanced nephelometric immunoassay(PENIA) from Dade Behring Co.. The reference ranges for Cys C in serum of 120 healthy individual were evaluated.Results The within-run CVs of the three reagents (A, B and C) were 3.08%-3.2%, 2.3%-4.15% and 1.38%-1.53% respectively.The total CV in A, B and C were 3.29%-3.44%, 2.65%-5.18% and 1.67%-1.69% respectively, lower than the stated.Limits of quantitative determination (LOQ) of the three reagents were 0.41, 0.23 and 0.07 mg/L, basically meeting the testing requirement.The linearity range was 0.22-7.26 mg/L(r=0.996), 0.20-7.72 mg/L(r=0.999)and 0.20-7.62 mg/L(r=0.997)in the three reagents, which demonstrated a sound linear correlation. For interference tests, no remarkable interference (<±10%) of reagent C was detected when bilirubin≤684 μmol/L, hemoglobin≤9.7 g/L and Chyle turbidity≤6 200 FTU; and no significant interference of reagent B was found when bilirubin≤684 μmol/L, hemoglobin≤6.79 g/L and Chyle turbidity≤6 200 FTU; when bilirubin≤684 μmol/L, hemoglobin≤4.85 g/L and Chyle turbidity≤1 240 FTU reagent A was not interfered significantly. The comparison afte and before the high-speed centrifugation reveals that the average percentage of bias for reagents A, B, C measured Cys C in chylous serum samples of patients was -8.31%, 1.52%, 1.32%, respectively.In method comparison tests, the regression equations of the three reagents compared with Dade Behring PENIA Cys C reagent were as follows:Y=0.787X+0.492 (R2=0.976), Y=1.098X+0.137 (R2=0.982) and Y=1.037X+0.249 (R2=0.996), respectively. Agreement rates of the high Cys C in reagent A, B, C and Dade Behring Cys C reagent were 80% (Kappa=0.615,P=0.000), 100% (Kappa=1.000,P=0.000), 91.2% (Kappa=0.824,P=0.000); While for reference range of preliminary clinical assessment, diagnosis coincidence rate of reagent A increased to 98.8% (Kappa=0.974,P=0.000). Conclusions When used in automatic biochemical analyzer, the three Cys C reagent with PETIA showed high precision,sensitivity, and sound correlation with Dade Behring PENIA reagents.The three reagents are all able to meet clinical test requirements, nevertheless, anti-interference capability were diffierent and the reference range should be further validated.