郑州大学学报（医学版）2017，Vol.52Issue(5)：541-544,4.DOI:10.13705/j.issn.1671-6825.2017.05.006
肠道病毒71型荧光定量PCR方法的比较与评价
Comparison and evaluation of different fluorescent quantitative PCR methods for enterovirus 71
摘要
Abstract
Aim:To determine copy number of the enterovirus 71( EV71) in clinical specimens by quantitative analysis , and compare 3 commercial TaqMan fluorescence quantitative PCR methods systematically and comprehensively .Methods:VP1 sequence of EV71 was inserted into cloning vector pMAL-c2X.The purified plasmid was used as the standard sample for quantitative detection of 105 clinical specimens collected from the special monitoring system of HFMD in Henan Province . Positive rate and CT value among the 3 commercial TaqMan fluorescence quantitative PCR methods from bioPerfectus Technol-ogies(method A), KINGHAWK(method B), and Beijing ABT(method C) were compared, and consistency was analyzed . Results:The recombinant plasmid pMAL-c2X was constructed successfully .Standard curve equation was Y=-3.35 X+47.49.For 105 specimens, positive rate of EV71 measured by method A, B, and C was 41.90%(44/105), 42.86%(45/105), and 41.90%(44/105), respectively, and the difference was not significant (P=0.987).Among positive specimens, copy number by method C(14.17 ±3.19) was less than method A(24.34 ±3.92) and B(25.08 ±3.94), and the difference was significant(P<0.05).For 3 methods, the limit of detection was 102, 10, and 102 copies; sensitivity was 93.62%, 95.74%, and 93.62%;specificity was 100.00%, 75.00%, and 100.00%.The result of 3 methods showed poor consisten-cy.Conclusion:EV71 fluorescence quantitative PCR kit for commercialization is suitable for the qualitative detection of clini -cal samples;however , it needs further study for the quantitative detection of EV 71.关键词
肠道病毒71型/荧光定量PCR/TaqMan探针Key words
enterovirus 71/fluorescent quantitative PCR/TaqMan probe分类
医药卫生引用本文复制引用
赵伊珂,卫海燕,黄学勇,程宁宁,潘静静,王若琳,许汴利,郭万申..肠道病毒71型荧光定量PCR方法的比较与评价[J].郑州大学学报（医学版）,2017,52(5):541-544,4.基金项目
国家自然科学基金资助项目 81573204 （）
河南省杰出青年资助项目 164100510008 （）
河南省科技厅项目 122102310268 （）
