Analysis of Positive Flow Cytometric Crossmatch in Organ Transplantation

Background:The results of human leukocyte antigen flow cytometry crossmatch (FCXM) are widely used when making transplant decisions. However, standardization/harmonization is required because the results vary between laboratories. A harmonized test method was reported to reduce interlaboratory variability. In this study, we evaluated the feasibility of establishing common cut-off values using a harmonized method in multicenter settings. Methods: Six laboratories participated in the study and conducted FCXM using a harmonized test method. Tests were performed using two donor cells and 25 negative sera samples. As a negative control (NC), laboratory NC (Lab NC) and a common NC (Korea Organ Donation Agency [KODA] NC) were included simultaneously. Median fluorescent intensity (MFI) ratios were calculated for each NC, and means ±2 standard deviation (SD) and ±3SD were estimated for the MFI ratio. Results: The suggested cut-off values based on the mean ±2SD and ±3SD of the MFI ratio data from 144 non-pronase T cell crossmatch after excluding one positive result were 1.6 and 1.9 using Lab NC and 1.3 and 1.5 using KODA NC, respectively. For pronase B cell crossmatch, values of 1.4 and 1.6 using Lab NC and 1.3 and 1.5 using KODA NC were obtained for the mean ±2SD and ±3SD of the MFI ratio, respectively. Inter-laboratory variations of nonpronase T cell crossmatch and pronase B cell crossmatch were less than 30% when using Lab NC and KODA NC. Conclusions: Variations in FCXM between laboratories were within the tolerable range when the harmonized method and same negative sera were applied. Therefore, applying common cut-off values with a standardized protocol may be feasible in multicenter settings.

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Feasibility of Establishing a Common Cut-Off of Human Leukocyte Antigen Flow Cytometry Crossmatch Using Harmonized Protocols in a Multicenter Setting

Bang

Park

Song

et al. 2022

J Lab Med Qual Assur

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A Questionnaire Survey of HLA Crossmatch Tests in Korea (2015)

Kang

Park

2017

Lab Med Online

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Evaluation of Flow Cytometric Crossmatch Results in Comparison with Donor-specific Antibodies Detected by Luminex-PRA Tests in Organ Transplantation Patients

Kim

Han

Hyun

et al. 2012

Korean J Transplant

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Background: Two of the most sensitive methods for detecting donor-specific HLA antibodies (DSAs) are solid phase panel reactive antibody (PRA) assay using Luminex platform (Luminex-PRA), and a cell-based flow cytometric crossmatch (FCXM) test. We evaluated FCXM results in relation to DSAs detected by the Luminex-PRA method in solid organ transplantation candidates or post-transplant follow-up patients.Methods: A total of 171 donor-recipient pairs were evaluated by Luminex-PRA (LIFECODES Class I and Class II ID kits; Gen-Probe, USA) and FCXM (T-and B-cells) tests. DSA levels were analyzed using a sum of median fluorescence intensity (MFI) values, and FCXM results were analyzed using MFI ratios.Results: Class I and II DSAs were detected in 11.7% (20/171) and 11.1% (19/171) of tested sera, respectively. T-FCXM was negative in 97.4% (147/151) of Class I DSA negative sera, and B-FCXM was negative in 99.3% (137/138) of Class I and II DSA negative sera. T-FCXM was positive in 91.7% (11/12) of sera with moderate to strong Class I DSAs and B-FCXM was positive in 88.9% (16/18) of sera with moderate to strong Class II and/or Class I DSAs in the evaluation of sensitivities of FCXM in relation to DSA. There were significant correlations between FCXM ratios and DSA levels for both T-FCXM (P=0.008) and B-FCXM (P＜0.001).Conclusions: The FCXM results correlated well with the DSAs detected by the Luminex-PRA method. The specificities of Tand B-FCXM in relation to DSAs were high (＞97%) and the sensitivities of T-and B-FCXM were satisfactory (＞88%) in detecting moderate to strong DSAs.

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Analysis of Positive Flow Cytometric Crossmatch in Organ Transplantation

Cited by 3 publications

References 19 publications

Feasibility of Establishing a Common Cut-Off of Human Leukocyte Antigen Flow Cytometry Crossmatch Using Harmonized Protocols in a Multicenter Setting

Feasibility of Establishing a Common Cut-Off of Human Leukocyte Antigen Flow Cytometry Crossmatch Using Harmonized Protocols in a Multicenter Setting

A Questionnaire Survey of HLA Crossmatch Tests in Korea (2015)

Evaluation of Flow Cytometric Crossmatch Results in Comparison with Donor-specific Antibodies Detected by Luminex-PRA Tests in Organ Transplantation Patients

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