A novel double antibody sandwich-lateral flow immunoassay for the rapid and simple detection of hepatitis C virus

Abstract

The objective of this study was to screen for antigens of the hepatitis C virus (HCV) to establish a new double antibody sandwich-lateral flow immunoassay (DAS-LFIA) method for testing the presence of anti-HCV antibodies in human serum or plasma. A series of different recombinant HCV proteins in Escherichia coli cells were constructed, expressed, purified and the new DAS-LFIA strip was developed. The sensitivity and specificity of new the DAS-LFIA strip were evaluated by detecting 23 HCV-positive sera, a set of quality control references for anti-HCV detection that contain known amounts of anti-HCV antibodies, and 8 HCV-negative sera. A total of 300 clinical serum samples was examined by both the new DAS-LFIA strip and enzyme-linked immunosorbent assay (ELISA). Data were analyzed using SPSS 11.5 software. The sensitivity and specificity of the new DAS-LFIA strip were 100%. The lowest test line of the HCV DAS-LFIA strips was 2 NCU/ml. Additionally, the concordance between the new DAS-LFIA strip and ELISA methods was 94.33%. In conclusion, our new testing method is rapid, simple, sensitive and specifically detects the presence of anti-HCV antibodies in human serum or plasma. Therefore, it may be used for monitoring HCV.

Expression of Hepatitis C Virus Core and E2 antigenic recombinant proteins and their use for development of diagnostic assays

“…19 However our expressed truncated Core antigen showed strong reactivity in 100% of PCR-positive samples and can act as a potential diagnosis tool for HCV infection. These results are markedly similar with the previous study in which the expressed HCV Core antigen showed 100% reactivity of PCR positive sera.…”

Is rapid hepatitis C virus testing from corpses a screening option for index persons who have died after mass-casualty incidents in high-prevalence settings in the field?

“…The best results from rapid tests are usually achieved in standardised evaluation studies, leading to sensitivities and specificities >98%,22 28 29 even for rapid multiplexed immunoassays (MBio Diagnostics, Boulder, Colorado, USA),30 and for various sample materials such as venous blood, finger-stick blood, serum, plasma or oral fluid (OraSure Technologies, Inc., Bethlehem, Pennsylvania, USA) 31. The OraQuick HCV test (OraSure Technologies, Inc.) in particular, suitable for blood and oral fluid, scored well in various evaluation studies 6 18 31–33.…”

Chromatographic Immunoassays: Strategies and Recent Developments in The Analysis of Drugs and Biological Agents

“…Another method combined chemiluminescent detection with traditional antibodies or nanobodies that were attached to magnetic beads and silica nanoparticles for use in flow-based sandwich immunoassays that could measure microcystin-LR or prealbumin [79,80]. Other chromatographic sandwich immunoassays have been described for IL-5, anti-HCV antibodies, potato virus X, cardiac troponin and myoglobin [81][82][83][84]. Sandwich immunoassays have also been applied to the determination of allergens, bacteria and toxins in food [85][86][87][88][89].…”

“…The utilization of alternative supports, such as affinity monoliths, is also expected to continue [110][111][112][113][114][115][116]. In addition, further research is anticipated in the development of miniaturized chromatographic immunoassays that can be employed in microfluidic devices and portable or disposable devices [19,[26][27][28]30,[82][83][84][85][86][87][88]100,113,116]. These develop ments, in turn, should increase the range of applications for which chromatographic immunoassays can be used in areas that include clinical chemistry, pharmaceutical testing, drug development and biomedical research [7,8].…”