ABI Quantistudio Dx实时定量PCR仪HBV-DNA检测的性能验证
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摘要
目的验证ABI Quantistudio Dx实时荧光定量PCR仪HBV-DNA检测系统的性能。方法参考GB/T 22576-2008和CNAS-CL36相关文件,ABI Quantistudio Dx实时荧光定量PCR仪检测患者标本、质控品及卫生部临检中心室间参考物质。从以下几个方面进行性能验证:精密度、正确度、线性范围、检出限等。结果 ABI Quantistudio Dx实时荧光定量PCR仪HBV-DNA检测系统的批内精密度(批内CV)分别为2. 09%(低浓度)、1. 3%(高浓度);批间精密度(批间CV)分别为3. 92%(低浓度)、2. 88%(高浓度);准确度评估以参加能力验证/室间质评的项目来判断,项目PT成绩为100%,通过;系统最低检出限为50 U/ml;线性范围验证结果显示,线性回归方程为Y=1. 0001X,R2=0. 9976。线性范围为1. 00×102～1. 00×108U/ml。结论 ABI Quantistudio Dx实时荧光定量PCR系统检测HBV-DNA性能良好,满足临床检测需要。
Objective To verify the performance of ABI Quantistudio Dx Real-time PCR HBV-DNA detection system.Methods According to CNAS-CL36 and GB/T 22576-2008,the analytical performance,such as precision,accuracy,limit of detection and linearity,was determined by ABI Quantistudio Dx real-time PCR system. Patient specimens,quality products,and reference materials from the National Center for Clinical Laboratories( NCCL) were employed for performance verification. Results The coefficient of variation( CV) of intra-assay of specimens with low concentration and high concentration of ABI Quantistudio Dx real-time PCR system was 2. 09% and 1. 3%,respectively,while the CV of inter-assay of specimens with a low concentration and high concentration was 3. 92% and 2. 88%,respectively. Analysis of reference materials from NCCL showed that test results were highly accurate. The detection limit of this system was 50 IU/ml. The linear regression equation was Y = 1. 0001 X,R2= 0. 9976. The linearity ranged from1. 00 × 102 to 1. 00 × 108 U/ml. Conclusions The performance of ABI Quantistudio Dx Real-time PCR HBV-DNA detection system meets the needs of clinical detection.
Objective To verify the performance of ABI Quantistudio Dx Real-time PCR HBV-DNA detection system.Methods According to CNAS-CL36 and GB/T 22576-2008,the analytical performance,such as precision,accuracy,limit of detection and linearity,was determined by ABI Quantistudio Dx real-time PCR system. Patient specimens,quality products,and reference materials from the National Center for Clinical Laboratories( NCCL) were employed for performance verification. Results The coefficient of variation( CV) of intra-assay of specimens with low concentration and high concentration of ABI Quantistudio Dx real-time PCR system was 2. 09% and 1. 3%,respectively,while the CV of inter-assay of specimens with a low concentration and high concentration was 3. 92% and 2. 88%,respectively. Analysis of reference materials from NCCL showed that test results were highly accurate. The detection limit of this system was 50 IU/ml. The linear regression equation was Y = 1. 0001 X,R2= 0. 9976. The linearity ranged from1. 00 × 102 to 1. 00 × 108 U/ml. Conclusions The performance of ABI Quantistudio Dx Real-time PCR HBV-DNA detection system meets the needs of clinical detection.
引文
[1]邢卉春,庞婷.从2015年中国《慢性乙型肝炎防治指南》看乙肝防治研究动态[J].武警医学,2016,27(5):433-436.
[2]中国国家标准化管理委员会. GB/T22576-2008医学实验室质量和能力的专用要求[S].北京:中国国家标准化管理委员会,2010.
[3]中国合格评定国家认可委员会. CNAS-CL02-A009:医学实验室质量和能力认可准则在分子诊断领域的应用说明[S].北京:中国合格评定国家认可委员会,2018.
[4] Clinical and laboratory standards institute. EP15-A2:User verification of performance for precision and trueness:approved guideline-second edition[S]. Wayne, PA,USA:NCCLS,2005.
[5] Clinical and laboratory standards institute. EP6-A:Evaluation of the linearity of quantitative measurement procedures:a statistical approach,approved guideline[S].Wayne,PA,USA:NCCLS,2003.
[6]辛娜,杨超,白跳艳,等.自建HBV-DNA实时荧光定量PCR检测系统性能验证方法及结果分析[J].山西医科大学学报,2016,47(4):365-368.
[7] Li M,Chen L,Liu L M,et al. Performance verification and comparison of Tian Long automatic hypersensitive hepatitis B virus DNA quantification system with roche CAP/CTM system[J]. World J Gastroenterol,2017,23(37):6845-6853.
[8]王娇,钱军,石英,等. Mx3000pTM实时荧光定量PCR仪HBV-DNA检测系统的性能验证[J].中国医药导报,2018,15(15):65-68.
[9]俞刚,陈瑜,张佳,等.罗氏Light Cycler 480实时荧光定量PCR系统检测HBV-DNA性能验证[J].标记免疫分析与临床,2016,23(10):1215-1225.
[10]巢薇. ABI 7300实时荧光定量PCR仪的性能评估[J].国际检验医学杂志,2015,36(4):494-495.
[11]胥明勇,朱华强. ABI 7500 HBV-DNA检测系统性能评价[J].实验与检验医学,2017,35(4):489-491.
[12]赵芹,辛青松,李峥嵘,等. ABI Vii A 7 Taqman HBV-DNA检测系统性能验证[J].国际检验医学杂志,2015,36(5):669-671.
[13]杨佳佳,张彦懿,刘华伟,等. HBV-DNA实时荧光定量PCR检测试剂盒性能验证[J].国际检验医学杂志,2018,39(2):213-217.
[14]高峰,高慧华,杨学文.脂血因素对荧光定量PCR检测乙肝病毒DNA的影响及解决措施[J].临床检验杂志,2007,25(5):359-360.
[15] Killeen A A,Long T,Souers R,et al. Verifying performance characteristics of quantitative analyticalsystems:calibration verification,linearity,and analytical measurement range[J]. Arch Pathol Lab Med,2014,138(9):1173-1181.
[2]中国国家标准化管理委员会. GB/T22576-2008医学实验室质量和能力的专用要求[S].北京:中国国家标准化管理委员会,2010.
[3]中国合格评定国家认可委员会. CNAS-CL02-A009:医学实验室质量和能力认可准则在分子诊断领域的应用说明[S].北京:中国合格评定国家认可委员会,2018.
[4] Clinical and laboratory standards institute. EP15-A2:User verification of performance for precision and trueness:approved guideline-second edition[S]. Wayne, PA,USA:NCCLS,2005.
[5] Clinical and laboratory standards institute. EP6-A:Evaluation of the linearity of quantitative measurement procedures:a statistical approach,approved guideline[S].Wayne,PA,USA:NCCLS,2003.
[6]辛娜,杨超,白跳艳,等.自建HBV-DNA实时荧光定量PCR检测系统性能验证方法及结果分析[J].山西医科大学学报,2016,47(4):365-368.
[7] Li M,Chen L,Liu L M,et al. Performance verification and comparison of Tian Long automatic hypersensitive hepatitis B virus DNA quantification system with roche CAP/CTM system[J]. World J Gastroenterol,2017,23(37):6845-6853.
[8]王娇,钱军,石英,等. Mx3000pTM实时荧光定量PCR仪HBV-DNA检测系统的性能验证[J].中国医药导报,2018,15(15):65-68.
[9]俞刚,陈瑜,张佳,等.罗氏Light Cycler 480实时荧光定量PCR系统检测HBV-DNA性能验证[J].标记免疫分析与临床,2016,23(10):1215-1225.
[10]巢薇. ABI 7300实时荧光定量PCR仪的性能评估[J].国际检验医学杂志,2015,36(4):494-495.
[11]胥明勇,朱华强. ABI 7500 HBV-DNA检测系统性能评价[J].实验与检验医学,2017,35(4):489-491.
[12]赵芹,辛青松,李峥嵘,等. ABI Vii A 7 Taqman HBV-DNA检测系统性能验证[J].国际检验医学杂志,2015,36(5):669-671.
[13]杨佳佳,张彦懿,刘华伟,等. HBV-DNA实时荧光定量PCR检测试剂盒性能验证[J].国际检验医学杂志,2018,39(2):213-217.
[14]高峰,高慧华,杨学文.脂血因素对荧光定量PCR检测乙肝病毒DNA的影响及解决措施[J].临床检验杂志,2007,25(5):359-360.
[15] Killeen A A,Long T,Souers R,et al. Verifying performance characteristics of quantitative analyticalsystems:calibration verification,linearity,and analytical measurement range[J]. Arch Pathol Lab Med,2014,138(9):1173-1181.