中国输血杂志2023，Vol.36Issue(12)：1107-1110,4.DOI:10.13303/j.cjbt.issn.1004-549x.2023.12.006
磁珠富集法提高外周血HIV DNA检测灵敏度的研究
Enhancing the sensitivity of HIV DNA detection:a magnetic bead-based method
摘要
Abstract
Objective To establish a magnetic bead enrichment strategy for the detection of human immunodeficiency vi-rus deoxyribonucleic acid(HIV DNA)in peripheral blood,and to verify the improvement of the sensitivity of this method for the detection of HIV DNA in HIV infected patients after early antiretrovital treatment(ART).Methods Peripheral whole blood was collected at 4 timepoints in one ART HIV window period(WP)patient.Peripheral blood mononuclear cells(PB-MCs)were isolated on a Ficoll gradient.CD4+T lymphocytes were enriched from total PBMCs by negative sorting.HIV DNA concentration in magnetic beads enriched group and whole blood group was detected by HIV DNA detection kit.Results CD4+T cells were isolated by magnetic beads and identified by FCM for purity at(96.4±2.6)%.The viability was(95.9±2.9)%,as demonstrated by trypan blue staining.The person on continued ART treatment in this study had significantly grea-ter reduction in HIV viral load and undetectable HIV plasma RNA at follow up timepoint 4.No HIV DNA was detected in the whole blood group at all 4 timepoints.The quantitative results of HIV DNA in the CD4+T lymphocyte group of the magnetic bead enrichment group were 73.4,429.3,137.1,449.9 copies/106 CD4+T cell′s respectively.Conclusion The magnetic bead enrichment method can be more sensitive in detecting the limit low copy HIV DNA in blood samples,and provide early confirmatory data for HIV WP infection and breakthrough infection after ART treatment.关键词
HIV DNA/早期感染/抗病毒治疗/磁珠法阴性分选/CD4+T细胞/人类免疫缺陷病毒Key words
HIV DNA/early infection/antiretroviral therapy drugs/magnetic bead negative sorting/CD4+T cells/HIV分类
医药卫生引用本文复制引用
王立林,邬林枫,赵方,李然,李彤,刘衡,朱蕊,曾劲峰..磁珠富集法提高外周血HIV DNA检测灵敏度的研究[J].中国输血杂志,2023,36(12):1107-1110,4.基金项目
2020年度广东省医学基金项目(A2020259),深圳市医学重点学科建设经费(SZXK070),深圳市医疗卫生三名工程项目(SZSM2023028),深圳市血液中心科研课题(SZBC202202) （A2020259）
