Microchip and capillary electrophoresis for quantitative analysis of hepatitis C virus based on RT-competitive PCR

Research output: Journal Publications and ReviewsRGC 21 - Publication in refereed journalpeer-review

17 Scopus Citations
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Author(s)

  • Kung-Chia Young
  • Hsiang-Mei Lien
  • Chun-Che Lin
  • Ting-Tsung Chang
  • Shu-Hui Chen

Detail(s)

Original languageEnglish
Pages (from-to)323-330
Journal / PublicationTalanta
Volume56
Issue number2
Online published27 Nov 2001
Publication statusPublished - 11 Feb 2002
Externally publishedYes

Abstract

A method to quantitatively perform reverse transcription-competitive PCR (RT-cPCR) of hepatitis C virus followed by both microchip and capillary electrophoretic separation and detection was described. In this method, HCV wild-type (WT) RNA extracted from serum was coretrotranscribed and coamplified with a constant amount of recombinant internal standard (IS) RNA which had the same primer binding region as the target RNA and was constructed by removing a centrally located 25-bp segment from the target template. A linear calibration curve was constructed by adding IS RNA at a constant concentration of 8000 copies μl-1 into a series of RNA target standards ranging from 400 to 106 copies μl-1. The amplified IS and target DNA were detected by both capillary and microchip electrophoresis via laser-induced fluorescence (LIF) using Cy5-labelled primer as the fluorescence probe. The method was further demonstrated for the quantitation of clinical patients with low, medium, and high viral titer and the results were found to be comparable to those determined by the commercial bDNA assay. © 2002 Elsevier Science B.V. All rights reserved.

Research Area(s)

  • Competitive PCR, Hepatitis C virus (HCV), Microchip electrophoresis, Quantitation

Citation Format(s)

Microchip and capillary electrophoresis for quantitative analysis of hepatitis C virus based on RT-competitive PCR. / Young, Kung-Chia; Lien, Hsiang-Mei; Lin, Chun-Che et al.
In: Talanta, Vol. 56, No. 2, 11.02.2002, p. 323-330.

Research output: Journal Publications and ReviewsRGC 21 - Publication in refereed journalpeer-review