PubMed · 13254924
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M GAGALA. 1955. [Coombs test].. https://pubmed.ncbi.nlm.nih.gov/13254924/
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HLA Class I antibody screening can be performed by flow cytometry using a mixture of 30 distinct bead populations each coated with the Class I antigen phenotype derived from different cell lines. In this study we compared the efficacy of Class I antibody screens done by flow cytometry beads with the antihuman globulin (AHG) method for patients awaiting cadaveric renal retransplantation. Class I panel reactive antibody (PRA) screening by flow cytometric beads of 21 regraft serum samples that had all been found to be negative by AHG DTT Class I PRA, revealed that 57.1% (12 of 21) had a flow Class I PRA of > or = 10%. Furthermore, when five regraft sera with an intermediate PRA were screened (mean AHG DTT PRA = 33.2 +/- 13%) the mean flow Class I PRA almost doubled (mean flow PRA = 72.4 +/- 10.2%) (p < 0.01). When active UNOS waiting list regraft candidates, after several months of screening the Class I PRA by flow beads, were divided into the three PRA categories based on their peak flow Class I PRA value (0-20%, 21-79% and > or = 80%), the incidence of a positive flow cross-match was 0%, 72% and 85% and the incidence of retransplantation was 60%, 22% and 10%, in each of these groups, respectively. These data provided our histocompatibility laboratory with the rationale to stop performing the AHG PRA and perform only the flow Class I PRA method for regraft candidates.
An international working party has conducted a study designed to select a suitable reference reagent for antihuman globulin, to replace those first made available in 1987. The chosen preparation contains levels of anti-IgG and anti-C3 (anti-C3c and anti-C3d) potency that are considered suitable to serve for reference when evaluating either polyspecific antihuman globulin reagents or those containing their separate monospecific components. The reference material is available in 2-ml freeze-dried aliquots from seven assigned distribution centres.
BACKGROUND: The gel test, developed by Lapierre in 1984, was designed to standardize antiglobulin testing while improving sensitivity and specificity of the method. PRINCIPLE: Anti-human serum immunoglobulin G (IgG) mixed with Sephadex G100 (gel phase) in a microtube traps red cell-IgG agglutination complexes during migration through the gel in a centrifugation step. Agglutination complexes are visibly detectable at various levels in the microtube as an inverse function of antibody coated on red cells. Unsensitized red cells form a cell pellet at the base of the microtube. OBJECTIVE: To determine if indirect anti-human globulin testing could be standardized and simplified by replacing the tube test with the gel test without compromising quality or increasing costs. SETTING: A medium-sized community hospital. RESULTS: In a blinded retrospective study, we used patient sera (n = 40), which included 10 positive specimens containing 18 known antibodies. Sixteen antibodies were detected and identified with the tube method (1 anti-D and 1 anti-C not detected). By the gel test, 18 antibodies were detected and identified. All negative samples showed 100% concordance. Favorable results were obtained in a nonblinded prospective correlation study (n = 121). Our technologists found the gel test easier to read and more reproducible and reliable than the tube method; they also found increased sensitivity for detecting weakly reacting antibodies. We successfully introduced the gel test into our laboratory as the standard method for indirect antiglobulin testing. Following implementation, improved personnel management was achieved. CONCLUSIONS: The gel test is a reliable and advantageous method and is appropriate for routine use for detection and identification of alloantibodies in a community hospital transfusion service laboratory.