两种国产荧光定量PCR试剂与Roche定量试剂的比较  

Comparison of two domestic fluorescent quantitative PCR reagents and Roche quantitative reagent

作　　者：杨鑫城 单幼兰[1] YANG Xincheng;SHAN Youlan(Department of Infectious Disease,the Second Affiliated Hospital of Chongqing Medical University,Chongqing 400010,China)

机构地区：[1]重庆医科大学附属第二医院感染病科,重庆400010

出　　处：《国际检验医学杂志》2023年第1期29-33,共5页International Journal of Laboratory Medicine

基　　金：重庆市自然科学基金面上项目(cstc2021jcyj-msxmX0059)。

摘　　要：目的探讨两种国产荧光定量聚合酶链反应(RT-qPCR)试剂以及Roche定量试剂检测乙型肝炎病毒核酸(HBV-DNA)结果之间的相关性,分析各方法之间的差异性,比较两种国产试剂的临床应用性能,选择更适合该实验室的试剂。方法收集用Roche定量试剂检测过的71例乙型肝炎患者血清,分别采用A、B两种试剂进行平行检测,然后对各检测结果进行比较,分析3种试剂之间的相关性和差异性;并对A、B试剂的最低检测限、最低定量限、线性范围、重复性精密度、正确度等性能指标进行验证。结果A、B试剂的检出率均高于Roche试剂,B试剂的检出率最高,且两种国产试剂定量结果与Roche试剂比较,差异无统计学意义(P>0.05)。A试剂与Roche试剂的符合率高于B试剂;A、B试剂的定量结果与Roche试剂均有良好的相关性,相关系数分别为0.896、0.908,差异均有统计学意义(P<0.05);A试剂的最低检测限为20 IU/mL,最低定量限为100 IU/mL,线性方程为Y=-0.996 X+8.925(R^(2)=0.999),在1.0×10^(2)~1.0×10^(8) IU/mL范围内线性良好,高浓度值变异系数为2.99%,低浓度值变异系数为4.70%,与Roche试剂的偏倚绝对值为6.69%;B试剂的最低检测限为20 IU/mL,最低定量限为20 IU/mL,线性方程为Y=-0.985 X+8.575(R^(2)=0.999),在2.0×101~1.0×10^(8) IU/mL范围内线性良好,高浓度值变异系数为0.73%,低浓度值变异系数为3.64%,与Roche试剂的偏倚绝对值为6.08%。结论B试剂核酸提取率较高,灵敏度和准确度也较高,重复性更好,且具有更宽的线性范围,但是自动化程度相对较低;总体来说,近年来一些国产HBV-DNA定量试剂性能逐渐优化,可以满足乙型肝炎患者的抗病毒和治疗监测的基本要求,其定量结果与Roche定量试剂有良好的相关性。通过比较,B试剂更适合该实验室。Objective To explore the correlation between two domestic fluorescent quantitative Polymerase Chain Reaction(RT-qPCR)reagents and Roche reagent in the detection of hepatitis B virus nucleic acid(HBV-DNA),analyze the differences between each method,compare the clinical application performance of the two domestic reagents,and select the reagent that is more suitable for the laboratory.Methods The serum of 71 patients with hepatitis B detected by Roche reagent were collected,A and B reagents were used for parallel detection,then the results were compared,and the correlation and difference between the three reagents were analyzed.The performance indexes of A and B reagent,such as the limit of detection,the limit of quantification,the linear range,the repeatability precision,accuracy,etc.were verified.Results The detection rates of reagent A and B were higher than that of Roche reagent,and the detection rate of reagent B was the highest,and there was no significant difference between the quantitative results two dommestic reagents with Roche reagent(P>0.05).The coincidence rate between reagent A and Roche reagent was higher than that of reagent B.The quantitative results of reagent A and B were well correlated with Roche reagent,and the correlation coefficients were 0.896 and 0.908,respectively,and differences were statistically significant(P<0.05).The limit of detection of reagent A was 20 IU/mL,and the limit of quantification of reagent A was 100 IU/mL,the linear equation was Y=-0.996 X+8.925(R^(2)=0.999).The linearity was good in the linarity range 1.0×10^(2)—1.0×10^(8) IU/mL.The coefficient of variation of high concentration value was 2.99%,the coefficient of variation of low concentration value was 4.70%,and the absolute value of bias with Roche reagent was 6.69%.The limit of detection of reagent B was 20 IU/mL,and the limit of quantification of reagent B was 20 IU/mL,the linear equation was Y=-0.985 X+8.57(R^(2)=0.999).The linearity was good in the linarity range 2.0×101—1.0×10^(8) IU/mL.The coefficient

关 键 词：荧光定量聚合酶链反应 Roche定量试剂 乙型肝炎病毒核酸 

分 类 号：R512.62[医药卫生—内科学]