Ann Lab Med.  2014 Jul;34(4):300-306. 10.3343/alm.2014.34.4.300.

Comparison of ABO Antibody Titers on the Basis of the Antibody Detection Method Used

• Kang SJ
• Lim YA
• Baik SY

• KMID: 1791938
• DOI: http://doi.org/10.3343/alm.2014.34.4.300

Abstract

BACKGROUND
Detection methods for ABO antibody (Ab) titers vary across laboratories, and the results are different depending on the method used. We aimed to compare titer values using different detection methods for the measurement of ABO Ab titers.
METHODS
For ABO Ab detection, pooled group A or B red blood cells (RBCs) were reacted with each of 20 sera from blood groups A, B, or O without dithiothreitol treatment. The room-temperature (RT) incubation technique and the indirect antiglobulin test (IAT) were used in the tube test and gel card test. Flow cytometry (FCM) was performed by using anti-IgM and anti-IgG Abs.
RESULTS
Regardless of the blood groups tested, the FCM assay with anti-IgM showed the highest titer compared to the tube test and gel card test with RT incubation in both. The tube test with IAT showed a higher titer than the gel card test with IAT (Gel-IAT) or FCM with anti-IgG in blood group A and B, while Gel-IAT showed the highest titer relative to the other tests, only for the anti-A Ab in blood group O.
CONCLUSIONS
There were significant differences in the titers depending on the detection method used, and each method showed a different detection capacity for each ABO Ab depending on the ABO blood group tested. Therefore, caution should be exercised in interpreting ABO Ab titer results, taking into consideration the detection method used and the blood group.

Keyword

ABO blood group system; Titer; Flow cytometry

MeSH Terms

ABO Blood-Group System/*immunology
Adult
Aged
Agglutination Tests/instrumentation/*standards
Antibodies/*analysis
Antibodies, Anti-Idiotypic/analysis
Erythrocytes/chemistry/metabolism
Female
*Flow Cytometry
Humans
Male
Middle Aged
Temperature
ABO Blood-Group System
Antibodies
Antibodies, Anti-Idiotypic

Figure

• 

  Fig. 1 Comparison of the effects of fixation of red blood cells (RBCs) on cell surface. Each sample was examined in the unfixed state (A) and after fixation with 0.05% glutaraldehyde (B). The same RBCs were used in both dot plots; Group B RBCs were reacted with A serum at room temperature and then incubated with fluorescein isothiocyanate (FITC)-labeled goat anti-human IgM.Abbreviations: SS, side scatter; FS, forward scatter.

• 

  Fig. 2 Comparison of ABO antibody titers between the tube test and the gel card test according to the blood group.*Mean difference in titer steps vs. gel card test; †P<0.001 vs. Gel card test; ‡P<0.01 vs. Gel card test.Abbreviations: Tube-RT, tube test with room temperature incubation method; Tube-IAT, tube test with indirect antiglobulin test; Gel-RT, gel card test with room temperature incubation method; Gel-IAT, gel card test with indirect antiglobulin test.

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