Nugent C Thomas, Dockter J, Bernardin F, Hecht R, Smith D, Delwart E, Pilcher C, Richman D, Busch M, Giachetti C
Gen-Probe Incorporated, San Diego, CA, USA.
J Virol Methods. 2009 Jul;159(1):10-4. doi: 10.1016/j.jviromet.2009.02.015. Epub 2009 Feb 24.
Peripheral blood mononuclear cells (PBMCs), saliva, seminal plasma, and dried blood spots were evaluated as specimen types for the APTIMA HIV-1 RNA Qualitative Assay (APTIMA HIV-1 Assay), which employs a target capture step to recover HIV-1-specific sequences from complex specimen types. Analytical sensitivity studies were carried out using samples that were either diluted or eluted with a buffered detergent and spiked with different concentrations of HIV-1 ranging from 1 to 10,000 copies/mL. PBMC samples spiked with HIV-1 had comparable analytical sensitivity to HIV-1 spiked plasma with a 95% limit of detection of 13.1 and 17.2 copies/mL, respectively. Analytical sensitivity in seminal plasma specimens diluted 1:5 and saliva diluted 1:2 was comparable to HIV-1 spiked dilution buffer alone. Whole blood and dried blood spot specimens spiked with HIV-1 had equivalent reactivity at 250 copies/spot (5000 copies/mL). However, the 95% limit of detection values were significantly different (293.7 copies/mL for whole blood and 2384 copies/mL for dried blood spot specimens). No significant effect on analytical sensitivity was observed when one HIV-1 positive dried blood spot punch was pooled with up to 9 HIV-1 negative dried blood spot punches. Together, these studies demonstrate that the APTIMA HIV-1 RNA Qualitative Assay can be used to process a diverse array of specimen types with minimal impact on analytical sensitivity for most specimen types.
外周血单核细胞(PBMCs)、唾液、精液和干血斑被评估为用于APTIMA HIV-1 RNA定性检测(APTIMA HIV-1检测)的样本类型,该检测采用靶标捕获步骤从复杂样本类型中回收HIV-1特异性序列。使用用缓冲去污剂稀释或洗脱并添加不同浓度(范围为1至10,000拷贝/毫升)HIV-1的样本进行分析灵敏度研究。添加HIV-1的PBMC样本与添加HIV-1的血浆具有相当的分析灵敏度,95%检测限分别为13.1和17.2拷贝/毫升。稀释1:5的精液样本和稀释1:2的唾液样本中的分析灵敏度与仅添加HIV-1的稀释缓冲液相当。添加HIV-1的全血和干血斑样本在250拷贝/斑(5000拷贝/毫升)时具有相同的反应性。然而,95%检测限值有显著差异(全血为293.7拷贝/毫升,干血斑样本为2384拷贝/毫升)。当将一个HIV-1阳性干血斑打孔样本与多达9个HIV-1阴性干血斑打孔样本混合时,未观察到对分析灵敏度有显著影响。总之,这些研究表明APTIMA HIV-1 RNA定性检测可用于处理多种样本类型,且对大多数样本类型的分析灵敏度影响最小。