|
|
|
| Performance evaluation of magnetic bead nucleic acid extraction in real-time fluorescence quantitative PCR detection system for plasma EBV-DNA |
| ZHENG Yuhong1 ZHONG Fengjin1 CHEN Yansong1 LIN Yingying1 XIAO Zhenzhou1 PAN Jianji2 LIN Shaojun2 GUO Qiaojuan2 CHEN Yan1▲ |
1.Department of Clinical Laboratory, Fujian Cancer Hospital & Fujian Medical University Cancer Hospital, Fujian Province, Fuzhou 350014, China;
2.Department of Radiotherapy, Fujian Cancer Hospital & Fujian Medical University Cancer Hospital, Fujian Province, Fuzhou 350014, China |
|
|
|
Abstract Objective To evaluate the application performance of magnetic bead nucleic acid extraction method in real-time fluorescence quantitative PCR detection for plasma Epstein-Barr virus (EBV) DNA. Methods Automated magnetic bead method was used to extract nucleic acid, and combined real-time fluorescence quantitative PCR assays for quantitative EBV-DNA. The detection performance of the method was evaluated by using the third-party reference materials. In addition, 100 patients with NPC were selected, and the plasma EBV-DNA was quantitatively detected by magnetic beads method and boiling method. The correlation between the two methods was analyzed. Results In the automated magnetic bead method-based real-time fluorescence quantitative PCR detection system for EBV-DNA, the repeatability precision of the low, medium and high were all <3/5 TEa (TEa = target ± 0.4 lg), and the intermediate precision were all < 4/5 Tea. In the range of 5.4×101 - 5.4×105 U/mL, there had good linear relationship. The minimum detection limit was about 3.334×101 U/mL. And the concentration of Hb interference in plasma was lower than 8.667 g/L, the results of magnetic bead test were not significantly affected. The detection results between magnetic bead method and boiling method were linearly related, and the qualitative coincidence rate was 83%. Conclusion The detection of plasma EBV-DNA by automated magnetic bead nucleic acid extraction method can achieve good detection performance.
|
|
|
|
|
|
| Cite this article: |
|
ZHENG Yuhong1 ZHONG Fengjin1 CHEN Yansong1 LIN Yingying1 XIAO Zhenzhou1 PAN Jianji2 LIN Shaojun2 GUO Qiaojuan2 CHEN Yan1▲. Performance evaluation of magnetic bead nucleic acid extraction in real-time fluorescence quantitative PCR detection system for plasma EBV-DNA[J]. 中国医药导报, 2017, 14(24): 13-16,24.
|
|
|
|
| URL: |
| http://www.yiyaodaobao.com.cn/EN/ OR http://www.yiyaodaobao.com.cn/EN/Y2017/V14/I24/13 |
[1] Le QT，Zhang Q，Cao H，et al. An international collaboration to harmonize the quantitative plasma Epstein-Barr virus DNA assay for future biomarker-guided trials in nasopharyngeal carcinoma [J]. Clin Cancer Res，2013，19（8）：2208-2215.
[2] 中国合格评定国家认可委员会.CNAS-CL36：2012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]. 2015.
[3] NCCLS. CLSI EP7-A2：Interference Testing in Clinical Chemistry； Approved Guideline-Second Edition [S]. 2005.
[4] NCCLS. CLSI EP17-A2：Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures；Approved Guideline-Second Edition [S]. 2012.
[5] NCCLS. CLSI EP6-A：Evaluation of the Linearity of Quantitative Measurement Procedures：A Statistical Approach；Approved Guideline [S]. 2003.
[6] Allen TC. Quality：walk the walk [J]. Arch Pathol Lab Med，2011，135（11）：1384-1386.
[7] Allen TC，Hammond ME，Robboy SJ. Quality and the college of american pathologists [J]. Arch Pathol Lab Med，2011， 135（11）：1441.
[8] Jennings L，Van DV，Gulley ML. Recommended principles and practices for validating clinical molecular pathology tests [J]. Arch Pathol Lab Med，2009，133（5）：743-755.
[9] Guzel O，Guner EI. ISO 15189 accreditation：Requirements for quality and competence of medical laboratories，experience of a laboratory Ⅰ [J]. Clin Biochem，2009，42（4-5）：274-278.
[10] Yanikkaya-Demirel G. ISO 15189 accreditation：Requirements for quality and competence of medical laboratories，experience of a laboratory Ⅱ [J]. Clin Biochem，2009，42（4-5）：279-283.
[11] 胡荣盛，薛静俊，朱振坤，等.自动核酸提取实时荧光定量PCR检测全血及血浆中EBV DNA载量的性能评价[J].现代实用医学，2013，25（8）：940-943.
[12] Thatcher SA. DNA/RNA preparation for molecular detection [J]. Clin Chem，2015，61（1）：89-99.
[13] 刘佳，徐军，王雪飞，等.三种HBV DNA提取方法对荧光定量PCR检测结果影响的比较[J].中华检验医学杂志，2008，31（7）：780-783.
[14] 陆佳飞，周科隆，王缦.磁珠法快速提取乙型肝炎病毒DNA方法的建立及其应用研究[J].中华检验医学杂志，2012，35（9）：843-850.
[15] Hayden RT，Yan X，Wick MT，et al. Factors contributing to variability of quantitative viral PCR results in proficiency testing samples：a multivariate analysis [J]. J Clin Microbiol，2012，50（2）：337-345.
[16] 李金明.临床实验室分子诊断的标准化[J].中华检验医学杂志，2006，29（6）：483-486. |
|
|
|
