Abstract: This invention provides sensitive nucleic acid hybridization assay methods for the detection of target animal nucleic acids in a biological sample, such as acellular fluids. The methods are particularly useful in early diagnosis of animal diseases, particularly chronic illnesses.

Abstract: A method for automatically selecting oligonucleotide hybridization probes for detecting a mutation causing a mismatch in a DNA duplex is based on a thermodynamic nearest-neighbour model for calculating a melting point, which is a temperature at which a predetermined percentage of a multitude of identical hybridized pairs of a certain hybridization probe and a DNA strand is in an annealed state. Using this model a first melting point of the respective probe hybridized with its complementary mutant DNA strand or opposite sense mutant DNA strand, a second melting point of the respective probe hybridized with its complementary wild-type DNA strand or opposite sense wild-type DNA strand, respectively, and a temperature difference between the first and second melting points are calculated for all possible hybridization probes. Then, the possible hybridization probes are ranked with regard to the calculated temperature difference.