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Abbott Laboratories Corporation

George H Shimer, Jr

John J Carrino

Keith C Backman

Karen Joy Shaw

George H Miller

Robert R Yocum

Roberta S Hare

The invention provides methods for identifying nucleic acids and polypeptides derived from fungi that are useful as antifungal targets for diagnosis and/or treatment of fungal diseases. The invention also provides isolated fungal-specific nucleic acids and polypeptides comprising antifungal targets. Methods for identifying antifungal agents that interact with and/or alter the function the antifungal targets are provided.

Method to identify fungal genes useful as antifungal targets

The present invention involves methods of improving the Ligase Chain Reaction (LCR.TM.) amplification schemes by modifying at least one probe end so that the probability of the probe contributing to spurious ligation and signal development is greatly reduced. Only after specific hybridization of the modified probe with true target are the modified ends 'corrected' by endonuclease IV in a target dependent fashion to allow participation of the probe in the enzymatic ligation reaction. Specific modifications include 3' phosphate blocking groups and nucleic acid overhangs containing an abasic site at the point of ligation. Further embodiments include probes modified to contain ribonucleotide moieties which, after amplification, can be cleaved by RNase to destroy the amplification products and reduce the risk of contamination.

Ligase chain reaction with endonuclease IV correction and contamination

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Boston, MA, United States of America

George H Shimer, Jr