Parker S P, Cubitt W D, Ades A E
Department of Virology, Camelia Botnar Laboratories, Hospital for Children, London, UK.
J Virol Methods. 1997 Nov;68(2):199-205. doi: 10.1016/s0166-0934(97)00127-4.
This study describes the development and evaluation of a cost effective test rationale for the detection of anti-HCV in dried blood spots. Samples were screened using an 'in house' IgG ELISA that incorporated the recombinant proteins c22-3, c200 and NS5. Confirmation of specific antibody to HCV was by a modification of the immunoblot RIBA 3.0. An extensive panel of well evaluated anti-HCV positive and negative samples from the UK and South Africa were used to assess the sensitivity and specificity of the two tests. One third of the anti-HCV positive samples had been typed. All anti-HCV positive samples were detected by the 'in house' screening EIA. Test/negative optical density ratios showed that more than 95% of reactive samples produced values greater than 5.0. Antibodies to HCV could be detected in a wide range of samples derived from asymptomatic and symptomatic patients and of different genotypes, with similar sensitivity. The presence of anti-HCV could be confirmed by RIBA 3.0 in samples with low reactivity but not in anti-HCV negative samples. Furthermore the immunoblot assay successfully increased specificity by screening out false reactive EIA samples that might occur in an epidemiological survey of a multi-ethnic population.
本研究描述了一种用于检测干血斑中抗丙型肝炎病毒(anti-HCV)的经济有效检测原理的开发与评估。使用一种包含重组蛋白c22-3、c200和NS5的“内部”IgG酶联免疫吸附测定(ELISA)对样本进行筛查。通过对免疫印迹RIBA 3.0进行改进来确认抗HCV特异性抗体。使用来自英国和南非的大量经过充分评估的抗HCV阳性和阴性样本评估这两种检测的敏感性和特异性。三分之一的抗HCV阳性样本已进行分型。所有抗HCV阳性样本均通过“内部”筛查酶免疫测定(EIA)检测到。检测/阴性光密度比值显示,超过95%的反应性样本产生的值大于5.0。在来自无症状和有症状患者的各种样本以及不同基因型的样本中均能检测到抗HCV抗体,且敏感性相似。RIBA 3.0可在低反应性样本中确认抗HCV的存在,但在抗HCV阴性样本中则不能。此外,免疫印迹测定通过筛选出在多民族人群流行病学调查中可能出现的假反应性EIA样本,成功提高了特异性。