Funding Information This work was supported by the Taiwan Food and Drug Administration and by National Taiwan University.
A reliable multiplex genotyping assay for HCV using a suspension bead array
Article first published online: 10 JUL 2014
© 2014 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Volume 8, Issue 1, pages 93–102, January 2015
How to Cite
Yang, Y.-C., Wang, D.-Y., Cheng, H.-F., Chuang, E. Y. and Tsai, M.-H. (2015), A reliable multiplex genotyping assay for HCV using a suspension bead array. Microbial Biotechnology, 8: 93–102. doi: 10.1111/1751-7915.12140
- Issue published online: 28 JAN 2015
- Article first published online: 10 JUL 2014
- Manuscript Revised: 27 MAY 2014
- Manuscript Accepted: 27 MAY 2014
- Manuscript Received: 4 APR 2014
- Taiwan Food and Drug Administration
- National Taiwan University
The genotyping of the hepatitis C virus (HCV) plays an important role in the treatment of HCV because genotype determination has recently been incorporated into the treatment guidelines for HCV infections. Most current genotyping methods are unable to detect mixed genotypes from two or more HCV infections. We therefore developed a multiplex genotyping assay to determine HCV genotypes using a bead array. Synthetic plasmids, genotype panels and standards were used to verify the target-specific primer (TSP) design in the assay, and the results indicated that discrimination efforts using 10 TSPs in a single reaction were extremely successful. Thirty-five specimens were then tested to evaluate the assay performance, and the results were highly consistent with those of direct sequencing, supporting the reliability of the assay. Moreover, the results from samples with mixed HCV genotypes revealed that the method is capable of detecting two different genotypes within a sample. Furthermore, the specificity evaluation results suggested that the assay could correctly identify HCV in HCV/human immunodeficiency virus (HIV) co-infected patients. This genotyping platform enables the simultaneous detection and identification of more than one genotype in a same sample and is able to test 96 samples simultaneously. It could therefore provide a rapid, efficient and reliable method of determining HCV genotypes in the future.