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Proceedings Paper

Rapid detection of IHNV by molecular padlock recognition and surface-associated isothermal amplification
Author(s): Erik L McCarthy; Teressa J. Egeler; Lee E. Bickerstaff; Mauricio Pereira da Cunha; Paul J. Millard
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Paper Abstract

RNA sequences derived from infectious hematopoeitic necrosis virus (IHNV) could be detected using a combination of surface-associated molecular padlock DNA probes (MPP) and rolling circle amplification (RCA) in microcapillary tubes. DNA oligonucleotides with base sequences identical to RNA obtained from IHNV were recognized by MPP. Circularized MPP were then captured on the inner surface of glass microcapillary tubes by immobilized DNA oligonucleotide primers. Extension of the immobilized primers by isothermal RCA gave rise to DNA concatamers, which were in turn bound by the fluorescent reporter SYBR Green II nucleic acid stain, and measured by microfluorimetry. Surface-associated molecular padlock technology, combined with isothermal RCA, exhibited high selectivity and sensitivity without thermal cycling. This technology is applicable to direct RNA and DNA detection, permitting detection of a variety of viral or bacterial pathogens.

Paper Details

Date Published: 9 November 2005
PDF: 7 pages
Proc. SPIE 5994, Chemical and Biological Sensors for Industrial and Environmental Security, 59940X (9 November 2005); doi: 10.1117/12.631072
Show Author Affiliations
Erik L McCarthy, Univ. of Maine (United States)
Teressa J. Egeler, Univ. of Maine (United States)
Lee E. Bickerstaff, Univ. of Maine (United States)
Mauricio Pereira da Cunha, Univ. of Maine (United States)
Paul J. Millard, Univ. of Maine (United States)

Published in SPIE Proceedings Vol. 5994:
Chemical and Biological Sensors for Industrial and Environmental Security
Arthur J. Sedlacek III; Steven D. Christesen; Roger J. Combs; Tuan Vo-Dinh, Editor(s)

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