Lab Med Qual Assur.
2021 Sep;43(3):152-161. 10.15263/jlmqa.2021.43.3.152.
Multicenter Verification of a Harmonized Test Method for Human Leukocyte Antigen Flow Cytometry Crossmatching
- Affiliations
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- 1Department of Laboratory Medicine, Severance Hospital, Yonsei University College of Medicine, Seoul, Korea
- 2Korea Organ Donation Agency Laboratory, Seoul, Korea
- 3Department of Laboratory Medicine, Korea University Guro Hospital, Seoul, Korea
- 4Department of Laboratory Medicine, Seoul St. Mary’s Hospital, College of Medicine, The Catholic University of Korea
- 5Department of Laboratory Medicine, Soonchunhyang University Seoul Hospital, Seoul, Korea
- 6Department of Laboratory Medicine, Yonsei University Wonju College of Medicine, Wonju, Korea
- 7Department of Laboratory Medicine and Genetics, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea
- KMID: 2520823
- DOI: http://doi.org/10.15263/jlmqa.2021.43.3.152
Abstract
- Background
Flow cytometry crossmatch (FCXM) is the most sensitive method currently used for human leukocyte antigen (HLA) crossmatches. The FCXM test methods and cut-off values used to determine the positive reaction vary significantly in different laboratories. To obtain comparable FCXM results from different laboratories and to use these results in deciding the compatibility between a donor and a recipient in organ transplantation, a standardized protocol needs to be established. In this study, we attempted to develop a harmonized test method and verify its performance with that of the different laboratories’ methods through a multicenter comparison.
Methods
The harmonized method was determined via a literature review and a survey that included seven laboratories. For the harmonized method, cell number, serum amount, pronase treatment for B cell FCXM, incubation temperature and time, and anti-human immunoglobulin G fluorescein isothiocyanate conjugate were standardized. Two trials of FCXM using one cell and three sera samples in each trial were performed, and the results of the laboratories’ methods versus those of the harmonized method were analyzed.
Results
Compared to the laboratory methods, the harmonized method showed increased agreement with consensus positive/negative results (75/84, 89.3% vs. 81/84, 96.4%) and a significantly decreased coefficient of variation among different laboratories in detecting the median fluorescence intensity (MFI) ratio values (88.3% vs. 55.2%, P=0.0003).
Conclusions
When the same protocol for the FCXM method is used, an increased consensus of positive/negative results and decreased variation of the MFI ratios from different laboratories can be obtained. Implementation of a harmonized and standardized protocol is needed in all domestic laboratories that perform the FCXM tests.
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