Method for detecting lupus anticoagulants
View Patent ↗Provided is the development of a convenient LA detection method in which even a sample derived from a patient who receives anticoagulant therapy of warfarin, heparin or the like, is not affected by the anticoagulant therapy, discrimination from the deficiency of blood coagulation factors is enabled, and healthy person's plasma is not used. The method for detecting lupus anticoagulant includes the following steps (A), (B) and (C): (A) a step of adding a buffer solution composition containing blood coagulation factors to each of a blood sample and a diluted sample of the blood sample before measurement or at the time of measurement of the blood coagulation time; (B) a step of measuring the blood coagulation times for the various samples of step (A); and (C) a step of comparing the blood coagulation times for the various samples obtained in step (B).
1. A method for detecting lupus anticoagulants, the method comprising:
(A) adding a composition comprising blood coagulation factors to a blood or plasma sample obtained from a subject that has been given warfarin or heparin, and a sample of the blood or plasma sample diluted of 1.1 to 3 times in a solution buffered at a pH of from 6 to 9 before measurement or at the time of measurement of blood coagulation time, wherein the blood coagulation factors are at least one selected from the group consisting of FII, FV, FVII, FVIII, FIX, FX, FXI and FXII,
wherein the composition is not a plasma or blood sample,
wherein the composition does not comprise phospholipids,
wherein the composition is buffered at a pH of from 6 to 9; and
wherein the composition comprises the blood coagulation factor in an amount of 0.01 U/ml to 2.0 U/ml;
(B) measuring the blood coagulation times for the blood sample and the diluted sample in an in vitro assay selected from the group consisting of measuring prothrombin time, measuring an activated partial thromboplastin time, measuring a diluted prothrombin time, measuring a kaolin clotting time, and measuring diluted Russell's viper venom time; and
(C) comparing the blood coagulation times for the blood sample and the diluted sample obtained in (B) to yield a ratio of the blood coagulation time of the diluted sample to the blood sample such that if the ratio is less than the same ratio measured from a healthy subjects blood or plasma indicates the presence of lupus anticoagulants.
2. The detection method according to claim 1 , wherein the blood sample is whole blood or plasma.
3. The detection method according to claim 1 , wherein the solution buffered at a pH of from 6 to 9 is added to the blood or plasma sample before the measurement of the blood coagulation time.
4. The detection method according to claim 1 , wherein the measuring of the blood coagulation time (B) comprises measuring activated thromboplastin time or dilute Russell's viper venom time.
5. The detection method according to claim 2 , wherein the solution buffered at a pH of from 6 to 9 is added to the blood or plasma sample before the measurement of the blood coagulation time.
6. The detection method according to claim 2 , wherein the measuring of the blood coagulation time (B) comprises measuring activated thromboplastin time or dilute Russell's viper venom time.
7. The detection method according to claim 3 , wherein the measuring of the blood coagulation time (B) comprises measuring activated thromboplastin time or dilute Russell's viper venom time.
8. The detection method according to claim 1 , wherein the blood coagulation is FII.
9. The detection method according to claim 8 , further comprising a second blood coagulation factor that is FX.
10. The detection method according to claim 9 , further comprising a second blood coagulation factor that is FV.
11. The detection method according to claim 1 , wherein the blood coagulation factors is FV.
12. The detection method according to claim 1 , wherein the blood coagulation factors is FX.
13. The detection method according to claim 11 , further comprising a second blood coagulation factor that is FX.
14. The detection method according to claim 1 , wherein the measuring of the blood coagulation time (B) comprising measuring activated thromboplastin time.
15. The detection method according to claim 1 , wherein the measuring of the blood coagulation time (B) comprises measuring dilute Russell's viper venom time.
16. The detection method according to claim 1 , wherein the solution buffered at a pH of from 6 to 9 is a HEPES buffer solution.