Evaluation of Platelet Alloimmunization by Filtration Enzyme-Linked Immunosorbent Assay.

Tzong-Shi Chiueh, Hsin-Yao Wang, Min-Hsien Wu, Yu-Shan Hsueh, Hui-Chu Chen
Author Information
  1. Tzong-Shi Chiueh: Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan City 333, Taiwan, China.
  2. Hsin-Yao Wang: Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan City 333, Taiwan, China. ORCID
  3. Min-Hsien Wu: Graduate Institute of Biochemical and Biomedical Engineering, Chang Gung University, Taoyuan City 333, Taiwan, China.
  4. Yu-Shan Hsueh: Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan City 333, Taiwan, China.
  5. Hui-Chu Chen: Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan City 333, Taiwan, China.

Abstract

The current methods for detecting antiplatelet antibodies are mostly manual and labor-intensive. A convenient and rapid detection method is required for effectively detecting alloimmunization during platelet transfusion. In our study, to detect antiplatelet antibodies, positive and negative sera of random-donor antiplatelet antibodies were collected after completing a routine solid-phase red cell adherence test (SPRCA). Platelet concentrates from our random volunteer donors were also prepared using the ZZAP method and then used in a faster, significantly less labor-intensive process, a filtration enzyme-linked immunosorbent assay (fELISA), for detecting antibodies against platelet surface antigens. All fELISA chromogen intensities were processed using ImageJ software. By dividing the final chromogen intensity of each test serum with the background chromogen intensity of whole platelets, the reactivity ratios of fELISA can be used to differentiate positive SPRCA sera from negative sera. A sensitivity of 93.9% and a specificity of 93.3% were obtained for 50 μL of sera using fELISA. The area under the ROC curve reached 0.96 when comparing fELISA with the SPRCA test. We have successfully developed a rapid fELISA method for detecting antiplatelet antibodies.

Keywords

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Grants

  1. CMRPVVK0051/Linkou Chang Gung Memorial Hospital
  2. CMRPG3K2451/Linkou Chang Gung Memorial Hospital

Word Cloud

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