High Diagnostic Accuracy of Antigen Microarray for Sensitive Detection of Hepatitis C Virus Infection

Abstract

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.

A Sol-Gel–Integrated Protein Array System for Affinity Analysis of Aptamer–Target Protein Interaction

“…A comparison with the 2-dimensional microarray system showed that sol-gel can immobilize proteins with a broad range up to milligram level with high activity (Kim et al, 2006;Kwon et al, 2008). Therefore, the sol-gel-integrated protein microarray appears to be ideal for measuring affinity, because sol-gel can house proteins with different concentration ranges.…”

Development of an ELISA-array for simultaneous detection of five encephalitis viruses

“…Moreover, when the volume of the clinical samples is limited and an increasing number of pathogens per sample needs to be tested, the detecting system should be high-throughput to allow detection of multiple pathogens simultaneously [6,20,21]. Multiple detection, easy to use, and affordability are requirements for detection methods in the clinical setting.…”

The Widely Used Diagnostics “DNA Microarray”-A Review

“…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, hence Kwon and co researchers developed a high diagnostic accuracy of antigen microarray for sensitive detection of hepatitis C virus infection and their method yielded significantly fewer false-positive results and the detection limit was reported to be 1000 times more sensitive than that of the ELISA and concluded that thus developed in expensive method will improve the specificity and sensitivity of large sample volume [43] . Ciccaglione and co-researchers [44] observed that microarray analysis created a common set of cellular genes modulated by different HCV replicon clones.…”