Hexagonal Phase Phospholipid
Neutralization, Hexagonal Phase Phospholipid
This test system is designed for the qualitative detection of lupus anticoagulants in plasma.6
Lupus anticoagulants are nonspecific inhibitors of phospholipid-dependent in vitro coagulation tests.6 The HPP assay takes advantage of the fact that many LA antibodies specifically recognize the HPP phospholipid configuration as an antigenic epitope. Addition of HPP to the reaction mixture serves to neutralize the inhibitory effect caused by LA antibodies and does not neutralize most factor-specific antibodies.9,10 The assay design includes several features that serve to improve the clinical utility of the test.7 The aPTT reagent used is diluted to reduce its phospholipid concentration and increase sensitivity to LA. The addition of normal plasma to the test system serves to correct for any clotting time prolongation caused by factor deficiencies in the patient plasma. This improves the specificity for LA by making the test relatively insensitive to factor deficiency. The assay reagent also contains a heparin neutralizer, which makes the test system insensitive to heparin levels up to 1 IU/mL. This assay can be used to detect lupus anticoagulants in patients receiving warfarin therapy.
Anticoagulant therapy may cause false-positive results. Plasma heparin levels >1 IU/mL may interfere with this test.7 Platelets are a rich source of phospholipid that can neutralize LA. Improper preparation of the platelet-poor plasma at collection reduces the sensitivity of this assay for LA. This test is subject to false-positive cross-reactivity with factor VIII inhibitors.8
Blue-top (sodium citrate) tube
Ideally, the patient should not be on anticoagulant therapy. Avoid warfarin (Coumadin®) therapy for two weeks prior to the test and heparin, direct Xa, and thrombin inhibitor therapies for about three days prior to testing. Do not draw from an arm with a heparin lock or heparinized catheter.
Citrated plasma samples should be collected by double centrifugation. The sample should be mixed immediately by gentle inversion at least six times to ensure adequate mixing of the anticoagulant with the blood. When noncitrate tubes are collected for other tests, collect sterile and nonadditive (red-top) tubes prior to citrate (blue-top) tubes. Any tube containing an alternate anticoagulant should be collected after the blue-top tube. Gel-barrier tubes and serum tubes with clot initiators should also be collected after the citrate tubes. Centrifuge for 10 minutes and carefully remove 2/3 of the plasma using a plastic transfer pipette, being careful not to disturb the cells. Deliver to a plastic transport tube, cap, and recentrifuge for 10 minutes. Use a second plastic pipette to remove the plasma, staying clear of the platelets at the bottom of the tube. Transfer the plasma into a frozen purple tube with screw cap. Freeze immediately and maintain frozen until tested.
Severe hemolysis; improper labeling; clotted specimen; specimen diluted with IV fluids; samples thawed in transit; improper sample type; sample out of stability