Method for nucleic acid amplification and detection using adhered probes

A method for the amplification and detection of a target nucleic acid comprising:

• A. amplifying a target nucleic acid using at least one biotinylated primer to form at least one amplified biotinylated target nucleic acid,
• B. contacting said amplified biotinylated target nucleic acid with a nucleic acid test element comprising a sealable support having disposed thereon a nucleic acid reagent composition comprising a mixture of:
• a. a nucleic acid reagent comprising:
• particles composed of a first polymer having a glass transition temperature of at least about 70.degree. C., said particles having an average diameter of from about 0.1 to about 3 micrometers, and
• an oligonucleotide covalently attached to said particles which is complementary to said target nucleic acid, and
• b. a water insoluble adhesive comprising a second polymer which has a glass transition temperature which is at least about 30.degree. C. less than the glass transition temperature of said first polymer,
• said polymeric adhesive being present in said composition at from about 1 to about 20 weight percent,
• wherein said sealable support has hydrophilic surface groups provided by a treatment selected from the group consisting of corona discharge treatment, chromic acid treatment and treatment with a radio frequency electromagnetic field in the presence of a reactive gas, and or said hydrophilic surface groups are provided by a hydrophilic subbing layer on which said composition is disposed,
• to form a water insoluble immobilized hybridized product of said amplified biotinylated target nucleic acid and said nucleic acid reagent on said element, and
• C. detecting the presence of said immobilized hybridized product by complexation of said amplified biotinylated target nucleic acid with a labeled conjugate of avidin as an indication of the presence of said target nucleic acid.