standard operating procedure for the direct antiglobulin test

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Provincial Blood Coordinating Program Standard Operating Procedure For Performing The Direct Antiglobulin Test. IgG C3 Positive Report result Report result Negative • add 1 drop IgG coated red cells • mix and centrifuge • read macroscopically • grade and record results • report result • incubate “C3” and “CTRL” tubes at room temperature 5 minutes • mix and centrifuge • read macroscopically and microscopically • grade and record result ▪ if still negative, add 1 drop C3 coated red cells ▪ mix and centrifuge ▪ read macroscopically ▪ grade and record results ▪ report result NOTE: If agglutination following the addition of the appropriate check cells is not detected the test is invalid and must be repeated 8.0 Reporting / Interpretation 8.1 Interpretation using Polyspecific AHG reagents: Polyspecific Control Test Result Interpretation AHG Negative Negative Negative Positive Negative Positive. Test with monospecific reagents. Positive Positive Unable to report. See note _______________________________________________________________________ This document may be incorporated into each Regional Policy/Procedure Manual. NL09-005 Version: 2.0 Effective Date: 2011-02-14 Page 8 of 12
Provincial Blood Coordinating Program Standard Operating Procedure For Performing The Direct Antiglobulin Test. 8.2 Interpretation using Monospecific antiglobulin reagents: Anti-IgG Anti- Control Test Result Interpretation Complement Negative Negative Negative Negative Positive Positive Negative Positive – IgG and Complement coating cells Positive Negative Negative Positive – IgG coating cells Negative Positive Negative Positive – Complement coating cells Negative TNP Negative Neonate specimens only: Negative Positive TNP Negative Neonate specimens only: Positive – IgG coating cells Positive Positive or Positive Unable to report See note TNP *TNP - Test not performed 9.0 Procedural Notes 9.1 Invalid tests 9.1.1 The addition of IgG control cells should demonstrate a 2+ reaction or the test is invalid and shall be repeated. 9.1.2 The addition of complement control cells should demonstrate a 1+ reaction or the test is invalid and shall be repeated. 9.2 False Negative test 9.2.1 Tests should be read immediately after centrifugation. Delay may cause bound IgG to dissociate from the red cells causing false negative results. 9.3 False Positive test 9.3.1 False positive test results due to in vitro coating with complement may occur if testing is performed on a clotted specimen. 9.4 Positive Control 9.4.1 A positive control may be due to strong cold agglutinin in the recipient’s serum/plasma. Wash recipient cells with 37°C and repeat DAT. 9.5 Mixed Field Reaction 9.5.1 Mixed Field reaction may indicate a transfusion reaction. 9.6 Weak Anti-complement Reactions 9.6.1 Weak anti-complement reactions may be enhanced if tubes containing polyspecific AHG /red cells or anti-complement / red cells are incubated for 5 minutes at room temperature after initial reading of the test. Centrifuge and read again. _______________________________________________________________________ This document may be incorporated into each Regional Policy/Procedure Manual. NL09-005 Version: 2.0 Effective Date: 2011-02-14 Page 9 of 12
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standard operating procedure for the direct antiglobulin test

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