Comparison of the tube test and column agglutination techniques for anti‐A/‐B antibody titration in healthy individuals

Abstract

Background and Objectives Determination of the anti-A/-B titre pre-and posttransplantation is beneficial for treatment selection. Currently, the recommended method for antibody titration is the tube test (TT) assay. Dithiothreitol (DTT) is used for IgM antibody inactivation. Recently, a fully automated antibody titration assay using the column agglutination technique (CAT) was developed (auto-CAT). Our aim was to compare the auto-CAT and TT techniques for ABO antibody titration, to evaluate the effectiveness of DTT-treated plasma for use with auto-CAT and to define the cut-off value for antibody titration by auto-CAT. Materials and MethodsWe enrolled 30 healthy individuals, including 10 each for blood types A, B and O. We performed antibody titre measurement using the TT technique and auto-CAT simultaneously. Auto-CAT uses the bead column agglutination technology. ResultsWith the auto-CAT cut-off value set to weak (w)+ with DTT treatment plasma, the concordance rate was 45%, and the weighted kappa value between TT and auto-CAT results was 0Á994 in all subjects. Furthermore, there was a significant positive correlation between the anti-A/-B titre results obtained using the TT technique and auto-CAT in all blood types. Moreover, a positive bias (falsely elevated end-points due to agglomeration of A/B cells) was not observed in auto-CAT testing using DTT-treated plasma.Conclusion Our results show that 1+ agglutination using the TT technique is equivalent to w+ agglutination obtained using auto-CAT. We recommend that DTT may be used with auto-CAT to measure antibody titres. Thus, we suggest that auto-CAT is useful for antibody titration in routine examination.

Comparative evaluation of the conventional tube test and column agglutination technology for ABO antibody titration in healthy individuals: a report from India

“…There are very few publications available in support of whether DTT can be used in CAT or whether DTT treatment of serum (or plasma) leads to falsepositive reactions by CAT. 10 As cases of ABO-incompatible transplants are gradually increasing in resource-constrained developing countries across the globe, it has become extremely important for immunohematology laboratories to validate an accurate method to estimate precise IgM and IgG levels of reactivity in ABO titers in patients as well as in potential healthy organ or stem cell donors.…”

Comparison of ABO isoagglutinin titres by three different methods in group O blood donors

“…Bhangle et al compared CTT with CAT for anti-A and anti-B titre estimation using 67 samples and concluded that CAT results are more sensitive than CTT [29]. Likewise, Matsura et al compared the titration by CTT with CAT using 30 samples and concluded that 1+ agglutination using the CTT technique is equivalent to weak agglutination obtained by the automated CAT [30]. They recommended that DTT may be used with automated CAT to measure antibody titres.…”

A standardized kodecyte method to quantify ABO antibodies in undiluted plasma of patients before ABO‐incompatible kidney transplantation

“…The level of ABO antibody is predominantly determined with plasma dilutions tested by either manual serology in tubes, or automated or semiautomated in cassettes (CAT) with both observing IgM direct agglutination and using antiglobulin reagents to detect IgG. We observed apparent differences in determination of antibody levels in both donors and patients, depending on the technology used.…”