Duke University Medical Center, Durham, NC, USA.
Blood Systems Research Institute, San Francisco, CA, USA.
J Immunol Methods. 2014 Jul;409:117-30. doi: 10.1016/j.jim.2014.01.004. Epub 2014 Jan 19.
The significant diversity among HIV-1 variants poses serious challenges for vaccine development and for developing sensitive assays for screening, surveillance, diagnosis, and clinical management. Recognizing a need to develop a panel of HIV representing the current genetic and geographic diversity NIH/NIAID contracted the External Quality Assurance Program Oversight Laboratory (EQAPOL) to isolate, characterize and establish panels of HIV-1 strains representing global diverse subtypes and circulating recombinant forms (CRFs), and to make them available to the research community. HIV-positive plasma specimens and previously established isolates were collected through a variety of collaborations with a preference for samples from acutely/recently infected persons. Source specimens were cultured to high-titer/high-volume using well-characterized cryopreserved PBMCs from National y donors. Panel samples were stored as neat culture supernatant or diluted into defibrinated plasma. Characterization for the final expanded virus stocks included viral load, p24 antigen, infectivity (TCID), sterility, coreceptor usage, and near full-length genome sequencing. Viruses are made available to approved, interested laboratories using an online ordering application. The current EQAPOL Viral Diversity panel includes 100 viral specimens representing 6 subtypes (A, B, C, D, F, and G), 2 sub-subtypes (F1 and F2), 7 CRFs (01, 02, 04, 14, 22, 24, and 47), 19 URFs and 3 group O viruses from 22 countries. The EQAPOL Viral Diversity panel is an invaluable collection of well-characterized reagents that are available to the scientific community, including researchers, epidemiologists, and commercial manufacturers of diagnostics and pharmaceuticals to support HIV research, as well as diagnostic and vaccine development.
HIV-1 变异体的显著多样性给疫苗开发以及开发用于筛选、监测、诊断和临床管理的敏感检测方法带来了严峻挑战。为了开发一组能够代表当前遗传和地理多样性的 HIV 代表株,NIH/NIAID 认识到需要建立一个 HIV 代表株的面板,NIH/NIAID 与外部质量保证计划监督实验室(EQAPOL)签订了合同,以分离、鉴定和建立代表全球不同亚型和循环重组形式(CRF)的 HIV-1 株的面板,并将其提供给研究界。通过与各种合作收集 HIV 阳性血浆标本和已建立的分离株,首选来自急性/近期感染者的样本。源标本在经过充分表征的冷冻 PBMC 中培养至高滴度/大容量,这些 PBMC 来自国家供体。将面板样本作为纯净培养上清液或稀释到纤维蛋白原血浆中储存。最终扩大病毒库存的特征包括病毒载量、p24 抗原、感染性(TCID)、无菌性、核心受体使用情况以及近乎全长基因组测序。通过在线订购应用程序向经过批准的有兴趣的实验室提供病毒。目前的 EQAPOL 病毒多样性面板包括 100 个代表 6 个亚型(A、B、C、D、F 和 G)、2 个亚亚型(F1 和 F2)、7 个 CRF(01、02、04、14、22、24 和 47)、19 个 URF 和来自 22 个国家的 3 种组 O 病毒的病毒样本。EQAPOL 病毒多样性面板是一组非常有价值的经过充分表征的试剂,可供科学界包括研究人员、流行病学家和诊断试剂和药物的商业制造商使用,以支持 HIV 研究以及诊断和疫苗开发。