Development of flow cytometry for detection and quantitation of red cell bound immunoglobulin G in autoimmune hemolytic anemia with negative direct Coombs test.

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dc.contributor.authorThedsawad, Anchalee
dc.contributor.authorTaka, Orathai
dc.contributor.authorWanachiwanawin, Wanchai
dc.date.accessioned2012-01-12T09:55:52Z
dc.date.available2012-01-12T09:55:52Z
dc.date.issued2011-12
dc.description.abstractAbout 2-10% of patients with warm-antibody autoimmune hemolytic anemia (WAIHA) exhibit a negative direct Coombs test (DAT), requiring more sensitive tests, including detection of RBC-bound immunoglobulins by flow cytometry, for diagnosis. In this study, the optimal conditions for detection and quantitation of RBC-bound IgG by flow cytometry were studied using blood samples from six patients with AIHA and two healthy individuals. Quantitation of RBC-bound IgG was performed using quantum simply cellular (QSC) beads coated with goat anti-mouse IgG antibodies. For detection of RBC bound IgG, a 60-minute incubation of all blood samples with 40 μl of 1:10 dilution of FITC-conjugated mouse anti-human IgG gave mean fluorescent intensity (MFI) values comparable to experiments using larger amounts or higher concentrations of the anti-human IgG. The acquired antibody binding capacity (ABC) values (or IgG molecules) for each QSC bead level, at 40 μl of 1:5 and 1:10 dilution of anti-human IgG for 60 minutes were close to the manufacturer-assigned ABC values. The IgG molecules per RBC in all six patients with positive DAT of 4+, 3+, 2+, 1+, trace and negative DAT were 31,725, 3,823, 1,753, 524, 260 and 88 respectively and in two healthy individuals with negative DAT they were 104 and 78.en_US
dc.identifier.citationThedsawad Anchalee, Taka Orathai, Wanachiwanawin Wanchai. Development of flow cytometry for detection and quantitation of red cell bound immunoglobulin G in autoimmune hemolytic anemia with negative direct Coombs test. Asian Pacific Journal of Allergy and Immunology, 2011 Dec; 29(4): 364-368.en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/136439
dc.language.isoenen_US
dc.source.urihttps://apjai.digitaljournals.org/index.php/apjai/article/viewFile/407/791en_US
dc.subjectred blood cellen_US
dc.subjectautoimmune hemolytic anemiaen_US
dc.subjectflow cytometryen_US
dc.subjectred cell bound immunoglobulin Gen_US
dc.subjectQuantum Simply Cellular (QSC) beaden_US
dc.titleDevelopment of flow cytometry for detection and quantitation of red cell bound immunoglobulin G in autoimmune hemolytic anemia with negative direct Coombs test.en_US
dc.typeArticleen_US
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