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.
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2017, Vol. 14 
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