不同生物活性基团修饰的纳米磁珠在核酸提取中的效能比较  

Comparison of the effectiveness of nanomagnetic beads modified with different bioactive groups in nucleic acid extraction

作　　者：曲业敏 邵丽莹[2] 袁晓燕 宋宇[1,2] 丛海燕 国东[2] 谢龙 迟翔宇 王明义 QU Yemin;SHAO Liying;YUAN Xiaoyan;SONG Yu;CONG Haiyan;GUO Dong;XIE Long;CHIXiangyu;WANG Mingyi(Weihai Weigao Biotechnology Co.,Ltd,Weihai,Shandong,China,264200;Weihai Municipal Hospi-tal Central Laboratory,Weihai,Shandong,China,264200;Research Center of Medical and Pharmaceutical Bioengineering,Ministry of Health,National and Regional Joint Engineering Laboratory for Clinical Medical Molecular Diagnostics,Guangdong Province Nucleic Acid Molecular Diagnostics Engineering Technology Re-search Center,Guangdong Provincial Clinical Medical Molecular Diagnostics Engineering Technology Center,DAAN Gene Co.,Ltd.,Guangzhou,Guangdong,510665)

机构地区：[1]威海威高生物科技有限公司,山东威海264200 [2]威海市立医院中心实验室,山东威海264200 [3]卫生部医药生物工程技术研究中心,临床医学分子诊断国家地方联合工程实验室,广东省核酸分子诊断工程技术研究中心,广东省临床医学分子诊断工程技术中心,广州达安基因股份有限公司,广东广州510665

出　　处：《分子诊断与治疗杂志》2024年第4期734-738,共5页Journal of Molecular Diagnostics and Therapy

基　　金：广东省重点领域研发计划项目(2019B110233003)。

摘　　要：目的评价不同生物活性基团修饰的纳米磁珠核酸提取效能,为临床应用及结果评价提供一些依据。方法通过不同生物活性基团修饰的纳米磁珠对检测乙型肝炎病毒的标本进行核酸提取,采用微量分光光度计(NanoDrop One~c)检测核酸的浓度及纯度,实时荧光定量聚合酶链式反应(RT-PCR)检测样本的CT值。结果在几种纳米磁珠提取法中,超顺磁性羧基纳米微粒提取的核酸浓度及纯度最高(浓度平均为953.3 IU/mL),其次是二氧化硅磁性微球与羧基微球混合纳米磁珠及纳米磁性氧化物MnFe_(2)O_(4)提取法。羟基修饰二氧化硅颗粒以及壳聚糖包覆的磁珠提取的核酸浓度较高,但纯度较差(A260/280及A260/230比值均超出标准范围)。对于同一样本,超顺磁性羧基纳米微粒提取的核酸扩增CT值为(21.28±0.36),核酸提取效率明显高于离心柱提取法,且具有统计学意义。二氧化硅磁性微球和羧基微球混合纳米磁珠、羟基修饰二氧化硅纳米磁珠及壳聚糖包覆的纳米磁珠核酸提取效率均低于离心柱提取法。此外,超顺磁性羧基纳米微粒的提取时间可缩短至15分钟。结论超顺磁性羧基纳米微粒核酸提取效率、精密度及核酸产物纯度最高,有较大的临床应用价值。Objective To evaluate the nucleic acid extraction efficiency of nanomagnetic beads modified with different bioactive groups to provide some basis for clinical application and outcome evaluation.Methods Nucleic acid was extracted from samples of the hepatitis B virus using nano⁃magnetic beads modi⁃fied with different bioactive groups.The concentration and purity of the nucleic acid were measureed using NanoDrop Onec,and the CT values of the samples were determined using real⁃time fluorescence quantitative polymerase chain reaction(RT⁃qPCR).Results Among several nanomagnetic bead extraction methods,the highest nucleic acid concentration and purity are achieved by superparamagnetic carboxyl nanoparticles(with an average concentration of 953.3 IU/mL).This is followed by silica magnetic microspheres,carboxyl micro⁃spheres mixed nanomagnetic beads,and the nano⁃magnetic oxide MnFe_(2)O_(4) extraction method.The nucleic ac⁃id concentration extracted by hydroxyl⁃modified silica particles and chitosan⁃coated magnetic beads is higher,but the purity is lower(with A260/280 and A260/230 ratios exceeding the standard range).For the same sam⁃ple,the CT value of nucleic acid amplification extracted by superparamagnetic carboxyl nanoparticles is(21.28±0.36),indicating significantly higher nucleic acid extraction efficiency compared to spin column extraction,with statistical significance.The nucleic acid extraction efficiency of silica magnetic microspheres,carboxyl mi⁃crospheres mixed nano⁃magnetic beads,hydroxyl⁃modified silica nano⁃magnetic beads,and chitosan⁃coated na⁃no⁃magnetic beads are all lower than that of the spin column extraction method.In addition,the extraction time of superparamagnetic carboxyl nanoparticles can be shortened to 15 minutes.Conclusion Superparamagnetic carboxyl nanoparticles have the highest efficiency in nucleic acid extraction,as well as precision and purity in nucleic acid products.They also hold great clinical application value.

关 键 词：纳米磁珠 实时荧光定量PCR 效能比较 

分 类 号：R440[医药卫生—诊断学]