Katsoulidou A, Moschidis Z, Sypsa V, Chini M, Papatheodoridis G V, Tassopoulos N C, Mimidis K, Karafoulidou A, Hatzakis A
Department of Hygiene and Epidemiology, Athens University Medical School, Athens, Greece.
Vox Sang. 2007 Jan;92(1):8-14. doi: 10.1111/j.1423-0410.2006.00857.x.
The Procleix Ultrio human immunodeficiency virus type 1 (HIV-1)/hepatitis C virus (HCV)/hepatitis B virus (HBV) (Ultrio) assay simultaneously detects HIV-1 RNA, HCV RNA and HBV DNA in individual blood donations. The main objective of the study was to assess the analytical and clinical sensitivity of the multiplex and discriminatory probe assays in samples with a low viral load.
The VQC HIV RNA genotype B, HCV RNA genotype 1 and HBV DNA genotype A standard dilutions were tested in 26 repeats. The probability of detection by Ultrio was compared with previously obtained data of the Procleix Duplex HIV-1/HCV assay on the same reference panels. A selection of 121 anti-HIV-1, 138 anti-HCV and 190 HBsAg positive samples from patients receiving antiviral therapy were tested. The majority of patient samples had a viral load below the detection limit of the diagnostic nucleic acid test assays, which made them suitable to evaluate the performance of the multiplex and discriminatory assays on yield cases with a similar low viral load.
The 95% and 50% detection end-points of the Ultrio assay along with the corresponding 95% confidence intervals are 53.7 (32.9-117.2) and 8.6 (6.2-12.1) geq/ml for HIV-1 RNA, 30.3 (19.0-62.4) and 5.2 (3.7-7.2) geq/ml for HCV RNA and 393.7 (147.9-6978) and 54.5 (22.4-143.8) geq/ml for HBV DNA. The analytical sensitivity of Ultrio expressed as a potency factor relative to previously obtained Duplex results on the same HIV-1 RNA and HCV-RNA standard dilutions was 1.09 (0.20-6.10) and 1.11 (0.21-5.89), respectively. The assay detected all 22 HIV-1 infected patients with viral load > 50 copies/ml, and 41 of 99 patients (41%) with viral load < 50 copies/ml, of which 23 (56%) were detected by the discriminatory assay. All 47 patients with HCV RNA load > 521 IU/ml and 10/91 polymerase chain reaction-negative patients with viral load < 50 IU/ml tested positive in Ultrio assay of which five were missed in the discriminatory test. The assay detected 53/55 HBV infected patients (96%) with viral load > 250 copies/ml and 108/135 patients (80%) with viral load < 250 copies/ml of which 17 (16%) were missed by the discriminatory test.
The new Procleix Ultrio assay is as sensitive as the Procleix Duplex assay for HIV-1 and HCV detection meeting the requirements of universal guidelines. The ability of the assay to detect HBV DNA in low viral load samples could be useful for screening blood. Inevitable negative results of discriminatory probe assays caused by stochastic sample variation will reduce the chance of recognizing low viraemic blood donors detected by individual donation nucleic acid test.
Procleix Ultrio 人类免疫缺陷病毒 1 型（HIV-1）/丙型肝炎病毒（HCV）/乙型肝炎病毒（HBV）（Ultrio）检测法可同时检测个体献血中的 HIV-1 RNA、HCV RNA 和 HBV DNA。本研究的主要目的是评估多重和鉴别探针检测法在低病毒载量样本中的分析灵敏度和临床灵敏度。
对 VQC HIV RNA B 基因型、HCV RNA 1 基因型和 HBV DNA A 基因型标准稀释液进行 26 次重复检测。将 Ultrio 的检测概率与之前在相同参考样本组上获得的 Procleix Duplex HIV-1/HCV 检测法的数据进行比较。对 121 例抗 HIV-1 阳性、138 例抗 HCV 阳性和 190 例 HBsAg 阳性的接受抗病毒治疗患者的样本进行检测。大多数患者样本的病毒载量低于诊断性核酸检测法的检测限，这使其适合评估多重和鉴别检测法在病毒载量相似的低产病例中的性能。
Ultrio 检测法的 95%和 50%检测终点以及相应的 95%置信区间分别为：HIV-1 RNA 为 53.7（32.9 - 117.2）和 8.6（6.2 - 12.1）geq/ml，HCV RNA 为 30.3（19.0 - 62.4）和 5.2（3.7 - 7.2）geq/ml，HBV DNA 为 393.7（147.9 - 6978）和 54.5（22.4 - 143.8）geq/ml。相对于之前在相同 HIV-1 RNA 和 HCV-RNA 标准稀释液上获得的 Duplex 结果，Ultrio 检测法以效价因子表示的分析灵敏度分别为 1.09（0.20 - 6.10）和 1.11（0.21 - 5.89）。该检测法检测出所有 22 例病毒载量 > 50 拷贝/ml 的 HIV-1 感染患者，以及 99 例病毒载量 < 50 拷贝/ml 患者中的 41 例（41%），其中鉴别检测法检测出 23 例（56%）。所有 47 例 HCV RNA 载量 > 521 IU/ml 的患者以及 91 例聚合酶链反应阴性且病毒载量 < 当病毒载量<50拷贝/ml患者中的10例在Ultrio检测中呈阳性，其中5例在鉴别检测中漏检。该检测法检测出 55 例病毒载量 > 250 拷贝/ml 的 HBV 感染患者中的 53 例（96%）以及 135 例病毒载量 < 250 拷贝/ml 患者中的 108 例（80%），其中 17 例（16%）在鉴别检测中漏检。
新型 Procleix Ultrio 检测法在检测 HIV-1 和 HCV 方面与 Procleix Duplex 检测法一样灵敏，符合通用指南的要求。该检测法在低病毒载量样本中检测 HBV DNA 的能力可能有助于血液筛查。鉴别探针检测法因随机样本变异导致的不可避免的阴性结果将减少识别个体献血核酸检测法检测出的低病毒血症献血者的机会。
