Rapid detection of IHNV by molecular padlock recognition and surface-associated isothermal amplification

Erik L McCarthy; Teressa J. Egeler; Lee E. Bickerstaff; Mauricio Pereira da Cunha; Paul J. Millard

doi:10.1117/12.631072

9 November 2005 Rapid detection of IHNV by molecular padlock recognition and surface-associated isothermal amplification

Erik L McCarthy, Teressa J. Egeler, Lee E. Bickerstaff, Mauricio Pereira da Cunha, Paul J. Millard

Author Affiliations +

Proceedings Volume 5994, Chemical and Biological Sensors for Industrial and Environmental Security; 59940X (2005) https://doi.org/10.1117/12.631072
Event: Optics East 2005, 2005, Boston, MA, United States

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.

Citation Download Citation

Erik L McCarthy, Teressa J. Egeler, Lee E. Bickerstaff, Mauricio Pereira da Cunha, and Paul J. Millard "Rapid detection of IHNV by molecular padlock recognition and surface-associated isothermal amplification", Proc. SPIE 5994, Chemical and Biological Sensors for Industrial and Environmental Security, 59940X (9 November 2005); https://doi.org/10.1117/12.631072

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