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Journal of Clinical Microbiology, September 1998, p. 2454-2459, Vol. 36, No. 9
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Cooperative Oligonucleotides Mediating Direct Capture of Hepatitis C Virus RNA from Serum

Deirdre O'Meara,1 Zhibing Yun,2 Anders Sönnerborg,2 and Joakim Lundeberg1,*

Department of Biochemistry and Biotechnology, Royal Institute of Technology (KTH),1 and Division of Clinical Virology, Karolinska Institute, Huddinge University Hospital,2 Stockholm, Sweden

Received 2 February 1998/Returned for modification 30 March 1998/Accepted 10 June 1998

A novel method for direct capture of hepatitis C virus (HCV) RNA from clinical samples has been developed. This approach takes advantage of the cooperative interactions between adjacently hybridized oligonucleotides. Here, this cooperative effect was combined with solid-phase technology, whereby a capture probe was covalently coupled to magnetic beads and a second probe, which anneals adjacent to the capture probe site, was prehybridized in solution to the target. When these contiguously hybridized probes were used for the extraction of HCV RNA from clinical samples, the capture efficiency was increased up to 25-fold in comparison to capture with a single probe. The applicability of this sample preparation assay was further investigated by performing a comparative study with both a conventional guanidinium extraction method and a commercial quantitative assay.

* Corresponding author. Mailing address: Department of Biochemistry and Biotechnology, Royal Institute of Technology (KTH), S-100 44 Stockholm, Sweden. Phone: 46 8 790 87 58. Fax: 46 8 24 54 52. E-mail: joakiml{at}biochem.kth.se.

Journal of Clinical Microbiology, September 1998, p. 2454-2459, Vol. 36, No. 9
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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