Titre plate and assay kit for detection of antibodies in human serum and their production and use

Claim 1

A method of producing an HTLV antigen-coated titer plate for the detection of antibodies to HTLV antigens in human sera which comprises: a) adding an aliquot of a diluted lot of HTLV antigens to the wells of a multi-well titer plate; b) storing said titer plate for sufficient time to permit a maximum amount of said HTLV antigens to adsorb onto said titer plate; c) removing any unadsorbed HTLV antigens from said titer plate; d) stabilizing said adsorbed HTLV antigens onto said plate; and e) storing said titer plate in a moisture-free container until ready to seal said plate.

Claim 2

A method of producing an HTLV antigen-coated titer plate for the detection of antibodies to HTLV antigens in human sera which comprises: a) adding an aliquot of a diluted lot of HTLV antigens to the wells of a multi-well titer plate; b) storing said titer plate for sufficient time to permit a maximum amount of said HTLV antigens to adsorb onto said titer plate; c) removing any unadsorbed HTLV antigens from said titer plate; d) stabilizing said adsorbed HTLV antigens onto said plate; and e) storing said titer plate in a moisture-free container until ready to seal said plate.

Claim 9

A method according to any one of claims 1 to 7 wherein said unadsorbed HTLV antigens are removed by rinsing said wells of said titer plate.

Claim 10

A method according to any one of claims 1 to 7 wherein said unadsorbed HTLV antigens are removed from said wells of said titer plate by standard suction techniques.

Claim 11

A method according to any one of claims 1 to 9 wherein the HTLV antigens are HTLV-III antigens.

Claim 17

A method for using an assay kit as claimed in any one of claims 11 to 14 which comprises the steps of: a) adding an aliquot of a serum diluent to each well of said titer plate which is to contain said positive control serum, said negative control serum, or human test serum; b) adding an aliquot of reconstituted positive control serum to at least one well of an HTLV antigen-coated titer plate which contains serum diluent; c) adding an aliquot of reconstituted negative control serum to at least one well of said titer plate which contains said serum diluent; d) adding an aliquot of human test serum to at least one well of said titer plate which contains said serum diluent; e) incubating said titer plate for such sufficient time as will permit a maximum amount of antibodies to the HTLV antigens to bind to said HTLV antigens, forming an antigen-antibody complex; f) rinsing each well of said titer plate with an appropriate rinsing solution to remove any unbound antibodies and to remove any excess of said human test serum; g) adding an aliquot of reconstituted and diluted anzyme-antibody conjugate to each well of said titer plate which contains aliquots of positive control serum, negative control serum, and human test serum; h) incubating said titer plate for such sufficient time as will allow said enzyme-antibody conjugate to bind to said antigen-antibody complex; i) rinsing said titer plate in an appropriate rinsing solution and removing the excess of said rinsing solution from said titer plate; j) adding an aliquot of a substrate solution to each well of said titer plate which contains said enzyme-antibody conjugate, said substrate solution being reactable with said enzyme-antibody conjugate; k) adding an aliquot of a stop solution to each well of said titer plate containing said substrate solution to stop any enzyme-substrate reaction; 1) measuring the absorbance of the reacted substrate by spectrophotometric methods; m) determining the presence of antibodies to HTLV antigens which may be present in said human test serum using the absorbance measurements obtained by said spectrophotometric methods.