Immunoassay of PIVKA-II
The present invention provides an immunoassay for specifically measuring with high sensitivity PIVKA-II in serum or plasma through antigen-antibody reaction by adding an animal serum containing thrombin and/or an antibody reacting with human fibrin-like related substances to the reagents. The immunoassay of the invention comprises the steps of adding thrombin and/or an antibody reacting with human fibrin-like related substances to the reagents, and measuring PIVKA-II in serum or plasma.
1. An immunoassay method for measuring PIVKA-II in a serum or plasma test sample, comprising the steps of:
(a) contacting the serum or plasma test sample with a first immunoassay reagent comprising anti-PIVKA-II antibodies and antibodies which specifically bind to a human fibrin or a human fibrinogen;
(b) contacting the resultant sample with a second immunoassay reagent comprising labeled antibodies which bind PIVKA-II, wherein the labeled antibodies are at least one antibody selected from the group consisting of anti-human prothrombin antibody, anti-F 1 antibody to the F 1 prothrombin fragment, anti-F 2 antibody to the F 2 prothrombin fragment and anti-(F 1 +F 2 ) antibody to the F 1 and F 2 prothrombin fragments; and
(c) measuring the labeled antibodies, thereby measuring PIVKA-II in the serum or plasma test sample.
2. The immunoassay method according to claim 1 , wherein the second immunoassay reagent further comprises thrombin.
3. The immunoassay method according to claim 2 , wherein the thrombin is a thrombin-containing animal serum and/or a purified thrombin.