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不同核酸提取方法在EB病毒和人巨细胞病毒检测中的效果

Effect of Different Nucleic Acid Extraction Methods in Detection of Epstein-Barr Virus and Human Cytomegalovirus
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摘要 目的:评价三种不同核酸提取方法在EB病毒(Epstein-Barr virus,EBV)和人巨细胞病毒(human cytomegalovirus,HCMV)核酸检测中的性能和临床应用价值。方法:采用手工柱提法、手工磁珠法及全自动磁珠法平行提取核酸,通过实时荧光定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qRT-PCR)测定EBV和HCMV的DNA,评估检测精密度、线性范围、最低检出限;选择2023年12月苏州大学附属第四医院的35份临床样本进行临床应用评价,同时评估手工磁珠法与全自动磁珠法核酸提取的线性关系。结果:3种方法在精密度方面的CV值均小于5%,在样本浓度为10^(2)~10^(7)拷贝/mL均具有良好的线性关系,最低检出限验证结果也均合格;在临床应用评价中,手工柱提法相较于其他两种方法检测阳性率更高,但差异无统计学意义(P>0.05),手工磁珠法与全自动磁珠法在EBV和HCMV样本的检测中具有良好的相关性。结论:三种核酸提取方法对EBV和HCMV样本的定量检测结果均符合要求,可根据不同样本量的检测需求选择合适的提取方法,以降低实验成本,提高检测效率。 Objective:To evaluate the performance and clinical application value of three different nucleic acid extraction methods in nucleic acid detection of Epstein-Barr virus(EBV)and human cytomegalovirus(HCMV).Method:Nucleic acids were extracted by manual column extraction method,manual magnetic bead method and automatic magnetic bead method.quantitative real-time polymerase chain reaction(qRT-PCR)was used to determine the DNA of EBV and HCMV.Evaluate detection precision,linear range,and minimum detection limit.Thirty-five clinical samples from the Fourth Affiliated Hospital of Soochow University in December 2023 were selected for clinical application evaluation,and the linear relationship between manual magnetic bead method and automatic magnetic bead method for nucleic acid extraction was evaluated.Result:The CV values of three methods in terms of precision were all less than 5%,and there was a good linear relationship in the sample concentration of 10^(2)-10^(7) copies/mL,the verification results of the minimum detection limit were also qualified;in the evaluation of clinical application,the positive rate of manual column extraction method was higher than those of the other two methods,but the difference was not statistically significant(P>0.05).There was a good correlation between manual magnetic bead method and automatic magnetic bead method in the detection of EBV and HCMV samples.Conclusion:The quantitative detection results of EBV and HCMV samples by three nucleic acid extraction methods all meet the requirements,the appropriate extraction method can be selected according to the detection requirements of different sample sizes to reduce the experimental cost and improve the detection efficiency.
作者 庄子 程建军 韩秀 周丽婷 ZHUANG Zi;CHENG Jianjun;HAN Xiu;ZHOU Liting(The Fourth Affiliated Hospital of Soochow University,Suzhou 215123,China;不详)
出处 《中外医学研究》 2024年第34期63-67,共5页 CHINESE AND FOREIGN MEDICAL RESEARCH
关键词 核酸提取 EB病毒 人巨细胞病毒 实时荧光定量聚合酶链式反应 Nucleic acid extraction Epstein-Barr virus Human cytomegalovirus Quantitative real-time polymerase chain reaction
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