Xi Dong, Luo XiaoPing, Lu QiangHua, Yao KaiLun, Liu ZuLi, Ning Qin
1Laboratory of Infectious Immunology, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030 Hubei P.R. China.
2Department of Pediatrics, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030 P.R. China.
J Nanopart Res. 2008;10(3):393-400. doi: 10.1007/s11051-007-9263-1. Epub 2007 Jul 17.
Gold-coated iron oxide nanoparticle Hepatitis B virus (HBV) DNA probes were prepared, and their application for HBV DNA measurement was studied. Gold-coated iron oxide nanoparticles were prepared by the citrate reduction of tetra-chloroauric acid in the presence of iron oxide nanoparticles which were added as seeds. With a fluorescence-based method, the maximal surface coverage of hexaethiol 30-mer oligonucleotides and the maximal percentage of hybridization strands on gold-coated iron oxide nanoparticles were (120 ± 8) oligonucleotides per nanoparticle, and (14 ± 2%), respectively, which were comparable with those of (132 ± 10) and (22 ± 3%) in Au nanoparticle groups. Large network aggregates were formed when gold-coated iron oxide nanoparticle HBV DNA gene probe was applied to detect HBV DNA molecules as evidenced by transmission electron microscopy and the high specificity was verified by blot hybridization. Our results further suggested that detecting DNA with iron oxide nanoparticles and magnetic separator was feasible and might be an alternative effective method.
制备了金包被的氧化铁纳米颗粒乙肝病毒(HBV)DNA探针,并研究了其在HBV DNA检测中的应用。在作为种子添加的氧化铁纳米颗粒存在下,通过柠檬酸还原四氯金酸制备金包被的氧化铁纳米颗粒。采用基于荧光的方法,六硫醇30聚体寡核苷酸在金包被的氧化铁纳米颗粒上的最大表面覆盖率和杂交链的最大百分比分别为每个纳米颗粒(120±8)个寡核苷酸和(14±2%),与金纳米颗粒组的(132±10)和(22±3%)相当。透射电子显微镜证明,当应用金包被的氧化铁纳米颗粒HBV DNA基因探针检测HBV DNA分子时,形成了大的网络聚集体,印迹杂交验证了其高特异性。我们的结果进一步表明,用氧化铁纳米颗粒和磁分离器检测DNA是可行的,可能是一种有效的替代方法。