Performance characteristics of the immunoglobulin G-capture BED-enzyme immunoassay, an assay to detect recent human immunodeficiency virus type 1 seroconversion
Performance characteristics of the immunoglobulin G-capture BED-enzyme immunoassay, an assay to detect recent human immunodeficiency virus type 1 seroconversion
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DOI：
10.1128/jcm.42.6.2623-2628.2004
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发表时间：
2004-06-01
影响因子：
9.4
通讯作者：
Parekh, BS
中科院分区：
文献类型：
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作者：
Dobbs, T;Kennedy, S;Parekh, BS
最近，我们开发了一种免疫球蛋白G（IgG）-捕获BED-酶免疫测定法（BED-CEIA），以确定近期人类免疫缺陷病毒（HIV）1型（HIV-1）血清转换用于发病率估计。我们已经建立了一个算法，其使用;开发质量控制试剂，以监测检测;并评估其性能的运行间，运行内，和操作员的变异性。对144个单个平板（涉及多个平板批次和多名操作员，超过一年）的分析表明，在0.5 - 2.0 OD单位的动态范围内，原始光密度（OD）值的变异系数（CV）在10%-15%之间;当OD归一化时，CV降至5%-10（OD-n; OD-n =样本OD/校准品OD）。对于OD 0.5的标本，运行内CV通常在5%至10%的范围内。多个平板批次（n = 6）和多名操作员（n = 7）之间的一致性水平相当高（R-2 > 0.9）。OD-n值小于或等于1.5的标本的初始和确证性检测结果的比较显示了高度相关性（R-2 = 0.92）;在两种模式下检测的615份标本中，566份（92%）保持了相同的分类（近期或长期感染）。分类改变的标本（n = 49）的值接近临界值（OD-n = 1.0），与预期一致。利用质控品和校准品试剂之间HIV IgG含量的两倍差异，通过使用质控图监测单个平板运行，将其纳入电子表格中进行数据输入和运行监测。这些信息为BED-CEIA成功转移到其他实验室以及使用BED-CEIA检测近期HIV血清转换和计算全球发病率估计值提供了基线数据。
Recently, we developed an immunoglobulin G (IgG)-capture BED-enzyme immunoassay (BED-CEIA) to identify recent human immunodeficiency virus (HIV) type 1 (HIV-1) seroconversion for use in incidence estimates. We have established an algorithm for its use; developed quality control reagents to monitor the assay; and evaluated its performance for interrun, intrarun, and operator variability. Analysis of 144 individual plates, which involved multiple plate lots and several operators over more than a year, indicated that the coefficients of variation (CVs) were between 10 and 15% for raw optical density (OD) values in the dynamic range between 0.5 and 2.0 OD units; the CVs decreased to 5 to 10% when the OD was normalized (OD-n; OD-n = specimen OD/calibrator OD). The intrarun CVs were generally in the range of 5 to 10% for specimens with ODs 0.5. The level of concordance between multiple plate lots (n = 6) and multiple operators (n = 7) was quite high (R-2 > 0.9). Comparison of the results of the initial and the confirmatory tests with specimens with OD-n values less than or equal to1.5 demonstrated a high degree of correlation (R-2 = 0.92); 566 (92%) of 615 of specimens tested in the two modes retained the same classification (recent or long-term infection). The values for those specimens with changed classifications (n = 49) were close to the cutoff (OD-n = 1.0), as expected. The twofold difference in the HIV IgG contents between the controls and the calibrator reagents was exploited to monitor individual plate runs by using a control plot, which was incorporated into the spreadsheet for data entry and run monitoring. This information provides baseline data for the successful transfer of BED-CEIA to other laboratories and the use of BED-CEIA for the detection of recent HIV seroconversion and the calculation of incidence estimates worldwide.