|本期目录/Table of Contents|

[1]郭 杰,曲 沛,李韦杰,等.丙型肝炎病毒核酸定量检测性能验证及评价[J].传染病信息,2019,05:394-398.
 GUO Jie,QU Pei,LI Wei-jie,et al.Performance verification and evaluation for hepatitis C virus nucleic acid quantitative detection[J].Infectious Disease Information,2019,05:394-398.
点击复制

丙型肝炎病毒核酸定量检测性能验证及评价(PDF)

《传染病信息》[ISSN:1007-8134/CN:11-3886/R]

期数:
2019年05期
页码:
394-398
栏目:
论著
出版日期:
2019-11-13

文章信息/Info

Title:
Performance verification and evaluation for hepatitis C virus nucleic acid quantitative detection
文章编号:
1007-8134(2019)05-0394-05
作者:
郭 杰曲 沛李韦杰周 宇郭晶晶焦炳欣王雅杰
 100015,首都医科大学附属北京地坛医院检验科(郭杰、曲沛、李韦杰、周宇、郭晶晶、焦炳欣、王雅杰)
Author(s):
GUO Jie QU Pei LI Wei-jie ZHOU Yu GUO Jing-jing JIAO Bing-xin WANG Ya-jie*
Department of Clinical Laboratory, Beijing Ditan Hospital, Capital Medical University, 100015, China
关键词:
HCV RNA定量检测实时荧光定量PCR性能验证
Keywords:
HCV RNA quantitative assay real-time fluorescence quantitative PCR performance verification
分类号:
R248.4
DOI:
10.3969/j.issn.1007-8134.2019.05.003
文献标识码:
A
摘要:
目的验证丙型肝炎病毒核酸定量检测试剂检出限发生改变时,能否达到厂家制定的分析指标。方法参照《临床实验室对商品定量试剂盒》 WS/T 420-2013的性能验证方案,采用 HCV RNA标准物质和首都医科大学附属北京地坛医院收集的不同浓度的临床标本,对丙型肝炎病毒核酸定量检测试剂的正确度、精密度、线性、检测限和抗干扰能力等参数进行方法学性能验证和评价。结果在正确度验证中,回归方程为 y=0.9881x-0.0972,R=0.998> 0.95,检测值与参考值高度相关。在精密度验证中,高浓度和低浓度标本的批内精密度 CV值(1.86%,2.64%)及批间精密度 CV值(1.44%,2.36%)均≤ 5%,符合要求。在线性验证中,在 2. 50E+2~ 5.00E+7 IU/ml分析测量线性范围良好。在检测限验证中,重复检测浓度 50 IU/ml样本 30次,其中 27次检出阳性,阳性率为 90%(27/30),符合临床要求。在抗干扰能力验证中,加入干扰物质混合后的血清与混合前血清定值比较,绝对偏差均< ±0.5 lg,检测结果符合临床需求。结论当试剂盒的检出限发生改变时,实验室应重点对该修改项的分析性能进行充分评估,以判断结果能否达到厂家制定的分析指标。同时依据卫生行业标准对该试剂其余性能指标进行验证。
Abstract:
ObjectiveTo validate whether the performance of hepatitis C virus nucleic acid quantitative assay kit matches the analytical performance promised by the manufacturer at changed detection limits. MethodsAccording to the performance validation program based on the WS/T420-2013 protocol of the Clinical Laboratory for Commercial Quantification Kit, the verification procedure was performed by using HCV RNA standards for reference and clinical specimens of different concentrations that were collected fromBeijing Ditan Hospital affiliated to Capital Medical University. The performance of hepatitis C virus nucleic acid quantitative assay kit was methodologically verified and evaluated through detecting the accuracy, precision, linearity, detection limit and anti-interferenceability of reagents. ResultsIn the accuracy verification, the regression equation was y=0.9881x-0.0972, R=0.998> 0.95, and the detected value was highly correlated with the reference value. In the precision verification, the intra-assay precision %CV value (1.86%, 2.64%) and the inter-assay precision %CV value (1.44%, 2.36%) of both high-concentration and low-concentration specimens were all ≤ 5%, which met the requirements. In the linearity verification, the linear range of 2.50E+2 to 5.00E+7 IU/ml was valid. In thedetection limit verification, samples at a concentration of 50 IU/ml were repeatedly measured 30 times, and 27 of the test results were positive, with a positive rate of 90% (27/30), which met the clinical requirements. In the anti-interference ability verification, the testvalues of serum samples with or without interfering substances were compared, and the absolute deviation of tests wasless than ±0.5 lg, which met the clinical requirements. ConclusionsWhen detection limits of an assay kit are revised, the laboratory should focus on a comprehensive evaluation of the analytical performance of the revised items, to judge whether the results match theanalytical performance claimed by the manufacturer. At the same time, all other performance indicators of the reagents are validatedaccording to the health industry standards.

参考文献/References

[1] Choo QL, Kuo G, Weiner AJ, et al. Isolation of a CDNA clone derived from a blood-bome non-A, non-B viral hepatitis viralgenome[J]. Science, 1989, 244(4902):359-362.
[2] Armstrong GL, Wasley A, Simard EP, et al. The prevalence ofhepatitis C virus infection in the United States, 1999 through 2002
[J]. Ann Intern Med, 144(10):705-714.
[3] Rustgi VK. The epidemiolgy of hepatitis C infection in the United States[J]. J Gastroentero, 42(7):513-521.
[4] Lauer GM, Walker BD. Hepatitis C virus infection[J]. N Engl J Med, 2001, 345(1):41-52.
[5] Cobb B, Pockros PJ, Vilchez RA, et al. HCV RNA viral load assessments in the era of direct-acting antivirals[J]. Am J Gastroenterol, 2013, 108(4):471-475.
[6] 中华人民共和国国家卫生和计划生育委员会 . 临床实验室对商品定量试剂盒分析性能的验证(WS-T 420-2013)[S]. 北京:中国标准出版社,2013.
[7] Clinical and Laboratory Standards Institute. EP9-A2 methodcomparison and bias estimation using patient samples; approvedguideline[S]. Wayne, PA: NCCLS, 2002.
[8] 国家食品药品监督管理局 . YY/T 1182-2010 核酸扩增检测用试剂(盒)[S]. 北京:中国标准出版社,2003.
[9] Clinical and Laboratory Standards Institute. EP6-A evaluation ofthe linearity of quantitative measurement procedures: a statisticalapproach: approved guideline[S]. Wayne, PA: NCCLS, 2003.
[10] 中华人民共和国国家质量监督检验检疫总局,中国国家标准化管理委员会 . ISO 18113-2:2009 体外诊断医疗器械制造商提供的信息(标示)第 2部分:专业用体外诊断试剂[S].北京:中国标准出版社,2014.

备注/Memo

备注/Memo:
[基金项目]首都医科大学附属北京地坛医院院内科研基金“育 苗计划”项目(DTYM201803)
[作者单位]100015,首都医科大学附属北京地坛医院检验科 (郭杰、曲沛、李韦杰、周宇、郭晶晶、焦炳欣、王雅杰)
[通信作者]王雅杰,E-mail:wangyajie@ccmu.edu.cn
*Correspondingauthor,E-mail:wangyajie@ccmu.edu.cn
更新日期/Last Update: 2019-11-13