Fonseca Bruna P F, Marques Christiane F S, Nascimento Lílian D, Mello Marcelle B, Silva Leila B R, Rubim Nara M, Foti Leonardo, Silva Edimilson D, Ferreira Antonio G P, Krieger Marco A
Laboratório de Tecnologia Diagnóstica, CEP 21045-900 Rio de Janeiro, RJ, Brazil.
Clin Vaccine Immunol. 2011 May;18(5):802-6. doi: 10.1128/CVI.00265-10. Epub 2011 Feb 23.
Hepatitis C virus (HCV) infection is a major burden to public health worldwide, affecting approximately 3% of the human population. Although HCV detection is currently based on reliable tests, the field of medical diagnostics has a growing need for inexpensive, accurate, and quick high-throughput assays. By using the recombinant HCV antigens NS3, NS4, NS5, and Combined, we describe a new bead-based multiplex test capable of detecting HCV infection in human serum samples. The first analysis, made in a singleplex format, showed that each antigen coupled to an individual bead set presented high-level responses for anti-HCV-positive reference serum pools and lower-level responses for the HCV-negative pools. Our next approach was to determine the sensitivity and specificity of each antigen by testing 93 HCV-positive and 93 HCV-negative sera. When assayed in the singleplex format, the NS3, NS4, and NS5 antigens presented lower sensitivity values (50.5%, 51.6%, and 55.9%, respectively) than did the Combined antigen, which presented a sensitivity of 93.5%. All antigens presented 100% specificity. These antigens were then multiplexed in a 4-plex assay, which resulted in increased sensitivity and specificity values, performing with 100% sensitivity and 100% specificity. The positive and negative predictive values for the 4-plex assay were 100%. Although preliminary, this 4-plex assay showed robust results that, aligned with its small-sample-volume requirements and also its cost- and time-effectiveness, make it a reasonable alternative to tests currently used for HCV screening of potentially infected individuals.
丙型肝炎病毒（HCV）感染是全球公共卫生的一项重大负担，影响着约3%的世界人口。尽管目前HCV检测基于可靠的测试，但医学诊断领域对廉价、准确且快速的高通量检测方法的需求日益增长。通过使用重组HCV抗原NS3、NS4、NS5及组合抗原，我们描述了一种新型的基于微珠的多重检测方法，该方法能够检测人血清样本中的HCV感染。首次以单重形式进行的分析表明，与单个微珠组偶联的每种抗原对HCV抗体阳性参考血清库呈现高水平反应，而对HCV阴性血清库呈现低水平反应。我们接下来的方法是通过检测93份HCV阳性血清和93份HCV阴性血清来确定每种抗原的敏感性和特异性。以单重形式检测时，NS3、NS4和NS5抗原的敏感性值较低（分别为50.5%、51.6%和55.9%），而组合抗原的敏感性为93.5%。所有抗原的特异性均为100%。然后将这些抗原用于四重检测中进行多重检测，结果敏感性和特异性值均有所提高，检测敏感性和特异性均为100%。四重检测的阳性预测值和阴性预测值均为100%。尽管该四重检测尚处于初步阶段，但其结果可靠，并符合小样本量要求以及成本效益和时间效益，使其成为目前用于对潜在感染个体进行HCV筛查的检测方法的合理替代方案。
