High diagnostic accuracy of antigen microarray for sensitive detection of hepatitis C virus infection
High diagnostic accuracy of antigen microarray for sensitive detection of hepatitis C virus infection
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DOI：
10.1373/clinchem.2007.090464
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发表时间：
2008-02-01
影响因子：
9.3
通讯作者：
Kim, Soyoun
中科院分区：
文献类型：
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作者：
Kwon, Jung-ah;Lee, Hyeseon;Kim, Soyoun
背景：丙型肝炎病毒（HCV）可通过输血传播。筛查ELISA是最广泛使用的HCV诊断方法，有时会产生假阳性和假阴性结果，因此使用确证试验。这种二次检测是劳动密集型的且昂贵的，因此对于大规模血库筛查是不切实际的。因此，需要一种新的高准确度的大规模筛选方法，以实现对HCV的敏感性和特异性检测。方法：采用溶胶-凝胶材料，在96孔板中设计新型抗原芯片，用于HCV检测。每个单独的孔用4种不同的HCV抗原点样。我们使用这种新系统检测了154例以前通过ELISA检测HCV的患者血清样本（87例HCV阳性和67例HCV阴性）（HCV EIA 3.0，ABBOTT）。我们评估了我们的微阵列系统的检测限与使用系列10倍稀释的HCV阳性samples.RESULTS：我们的微阵列检测是可重复的，并显示更高的诊断准确性（特异性）（98.78%）比ELISA（81.71%）。我们的方法产生的假阳性结果明显少于ELISA。该方法的检出限比ELISA法灵敏1000倍。此外，我们发现这种新的检测技术是兼容的，目前采用的自动化方法用于ELISA.CONCLUSION：我们成功地应用了溶胶-凝胶为基础的蛋白质微阵列技术的筛选检测HCV诊断与验证性测试水平的准确性。这种新的、廉价的方法将提高大规模样本诊断的特异性和灵敏度。
BACKGROUND: Hepatitis C virus (HCV) can be transmitted through blood transfusion. Screening ELISA, the most widely used method for HCV diagnosis, sometimes yields false-positive and false-negative results, so a confirmatory test is used. This secondary testing is labor-intensive and expensive, and thus is impractical for massive blood bank screening. Therefore, a new massive screening method with high accuracy is needed for sensitive and specific detection of HCV.METHODS: With sol-gel material, we designed novel antigen microarray in 96-well plates for HCV detection. Each individual well was spotted with 4 different HCV antigens. We used this new system to test 154 patient serum samples previously tested for HCV by ELISA (87 HCV positive and 67 HCV negative) (HCV EIA3.0, ABBOTT). We assessed the detection limit of our microarray system with the use of serial 10-fold dilutions of an HCV-positive sample.RESULTS: Our microarray assay was reproducible and displayed higher diagnostic accuracy (specificity) (98.78%) than did the ELISA (81.71%). Our method yielded significantly fewer false-positive results than did the ELISA. The detection limit of our assay was 1000 times more sensitive than that of the ELISA. In addition, we found this novel assay technology to be compatible with the currently employed automated methods used for ELISA.CONCLUSION: We successfully applied the sol-gel-based protein microarray technology to a screening assay for HCV diagnosis with confirmatory test-level accuracy. This new, inexpensive method will improve the specificity and sensitivity of massive sample diagnosis.