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      Screening and Identification of Unexpected Red Cell Antibodies by Simultaneous LISS/Coombs and NaCl/Enzyme Gel Methods

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          Abstract

          We evaluated the clinical usefulness of simultaneous LISS/Coombs and NaCl/Enzyme testing using the gel method for screening and identification of unexpected antibodies in 15,014 samples. When unexpected antibodies were detected by either screening test, those antibodies were identified using both the LISS/Coombs and the NaCl/Enzyme gel test. The positive screening rates of the LISS/Coombs, NaCl/Enzyme, and combined tests (excluding 25 autoantibody cases) were 0.48%, 1.29%, and 1.39%, respectively. Among the 57 samples positive by both screening methods, the antibodies in 19.3% could be identified only by the NaCl/Enzyme method. Among the 137 samples positive only by NaCl/Enzyme screening, 74.5% showed positive results in antibody identification only by the NaCl/Enzyme test, although 7.3% were also positive in the LISS/Coombs test. The NaCl/Enzyme method thus showed about threefold higher detection rates than the LISS/Coombs method, especially in screening for Rh antibodies, and higher exact identification rates and discriminatory power for identifying mixed antibodies. Addition of the NaCl/Enzyme method to routine laboratory procedures may detect and identify considerable numbers of significant antibodies that might be missed if only the LISS/Coombs method is used.

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          Most cited references18

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          The gel test: a new way to detect red cell antigen-antibody reactions.

          A new process for the detection of red cell (RBC) antigen antibody reactions is described. It is applicable to most of the tests performed in blood group serology. The procedures are standardized and easy, and they provide clear and stable reactions that improve the interpretation of results. The process uses special microtubes filled with a mixture of gel, buffer, and reagent. Depending on the test to be carried out, the test uses a neutral gel containing no reagents (reagents are added to top of gel) or a specific gel containing reagents (e.g., antiglobulin serum or anti-A, -B, -D, etc.). A suspension of RBCs (for typing or the direct antiglobulin test) or a mixture of RBCs and serum (for reverse ABO typing or antibody characterization) is centrifuged through the gel under precise conditions. In negative reactions, the RBCs pass through the gel and pellet in the bottom of the tube, whereas, in positive reactions, they are trapped in the gel and the reaction may be read for hours afterwards. The test is easy to perform, sensitive, and reproducible. The antiglobulin tests can be performed without washing of the RBCs. There should be a reduction of risk from biohazardous materials.
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            Guidelines for compatibility procedures in blood transfusion laboratories.

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              Lack of clinical significance of "enzyme-only" red cell alloantibodies.

              In a retrospective study on samples from 10,000 recently transfused patients, 35 samples were found to contain an antibody that reacted with ficin-treated red cells but was not demonstrable by low-ionic-strength saline solution and indirect antiglobulin test (LISS-IAT). In those 35 patients, the specificity of the antibody was such that each patient would have been transfused with antigen-negative blood had the antibody reacted in LISS-IAT. Tests on red cells from the units already transfused showed that 19 patients had among them received, by chance, 32 antigen-positive and 74 antigen-negative units. The remaining 16 patients had among them received 57 units that were, again by chance, all antigen negative. One patient given antigen-positive blood suffered a delayed transfusion reaction; in two others the antibodies became LISS-IAT active after transfusion. However, similar changes to the LISS-IAT-active state were seen with two antibodies of patients given only antigen-negative blood. Also found in the 10,000 patients were 28 clinically insignificant antibodies, 77 sera in which the antibody was too weak to identify, and 216 autoantibodies that reacted only with ficin-treated red cells. These data support a belief, generally held in the United States but not necessarily elsewhere, that the use of protease-treated red cells for routine pretransfusion tests creates far more work than the accrued benefits justify.
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                Author and article information

                Journal
                J Korean Med Sci
                JKMS
                Journal of Korean Medical Science
                The Korean Academy of Medical Sciences
                1011-8934
                1598-6357
                August 2009
                29 July 2009
                : 24
                : 4
                : 632-635
                Affiliations
                [1 ]Department of Laboratory Medicine, College of Medicine, Inje University, Busan, Korea.
                [2 ]Paik Institute for Clinical Research, Inje University, Busan, Korea.
                Author notes
                Address for correspondence: Jeong Nyeo Lee, M.D. Department of Laboratory Medicine, College of Medicine, Inje University, 633-165 Gaegeum-dong, Busanjin-gu, Busan 614-110, Korea. Tel: +82.51-890-6862, Fax: +82.51-893-1562, jeong418@ 123456medimail.co.kr
                Article
                10.3346/jkms.2009.24.4.632
                2719182
                19654944
                14bbd627-cfa1-46e4-b529-0fe9493a2b43
                Copyright © 2009 The Korean Academy of Medical Sciences

                This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 30 March 2008
                : 19 September 2008
                Categories
                Original Article

                Medicine
                erythrocyte antigens,nacl/enzyme,coombs' test,gel test,unexpected antibody
                Medicine
                erythrocyte antigens, nacl/enzyme, coombs' test, gel test, unexpected antibody

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