Related Experiment Video
Updated: Aug 6, 2026
In Vitro and In Vivo Assessment of T, B and Myeloid Cells Suppressive Activity and Humoral Responses from Transplant Recipients
Published on: August 12, 2017
Comparison between enzyme-linked immunosorbent assay and cytotoxic cross-match procedures for detecting IgG
B Susskind1, R H Kerman, R Nelson
1Division of Immunology and Organ Transplantation, University of Texas Health Science Center, Houston, USA.
Insights
The Cross-Stat assay, an enzyme-linked immunosorbent assay method, shows high sensitivity and negative predictive value for detecting IgG anti-donor HLA antibodies compared to standard complement-dependent cytotoxicity cross-match (CDC XM) methods. Further clinical relevance testing is recommended.
Area of Science:
- Transplantation immunology
- Immunodiagnostics
Background:
- Complement-dependent cytotoxicity cross-match (CDC XM) has limitations including complement requirement, need for viable cells, detection of non-HLA antibodies, and subjective scoring.
- Cross-Stat, an enzyme-linked immunosorbent assay (ELISA) based cross-match, was developed to overcome these limitations in detecting IgG anti-donor HLA antibodies.
Purpose of the Study:
- To compare the performance of the Cross-Stat assay with the standard anti-human globulin (AHG)-enhanced CDC XM procedure.
- To evaluate the sensitivity, specificity, and predictive values of the Cross-Stat assay in detecting anti-donor HLA antibodies.
Main Methods:
- A comparative study involving 524 sera from 230 transplant candidates and 51 cadaveric donors.
- Evaluation of results obtained from Cross-Stat and AHG-enhanced CDC XM procedures.
Main Results:
- A significant correlation (P<0.001) was observed between Cross-Stat and AHG-CDC IgG XM results.
- Cross-Stat demonstrated 100% sensitivity, 93% specificity, 73% positive predictive value, and 100% negative predictive value relative to AHG-CDC XM.
- Flow cytometry supported Cross-Stat results in cases of discrepancy, suggesting greater sensitivity.
Conclusions:
- The Cross-Stat assay offers theoretical advantages over CDC XM for detecting IgG anti-donor HLA antibodies.
- Despite a 15% indeterminate sample rate, the high sensitivity and NPV warrant further investigation into the clinical relevance of ELISA-based XM methods.
Background:
Disadvantages inherent to complement-dependent cytotoxicity cross-match (CDC XM) methods are the requirements for complement and viable target cells, detection of antibodies (Abs) against non-HLA antigens, and subjective scoring. Cross-Stat (SangStat Medical Corp., Menlo Park, CA), a recently developed enzyme-linked immunosorbent assay XM procedure for the detection of IgG anti-donor HLA Abs, is theoretically devoid of these flaws.
Methods:
We compared results of Cross-Stat and our standard anti-human globulin (AHG)-enhanced CDC XM procedure on 524 sera from 230 transplant candidates, which were evaluated against 51 cadaveric donors.
Results:
There was a significant correlation between AHG-CDC IgG XM and Cross-Stat results (P<0.001). For false negative sera, repeat AHG-CDC IgG XMs were still positive after platelet absorption, indicating that the Abs present were either non-HLA Abs or anti-HLA class II. Flow cytometry testing of false positive sera usually (42/62) substantiated Cross-Stat results, indicating that the discrepancy with AHG-CDC IgG XM is caused by greater sensitivity of Cross-Stat. Relative to the AHG-CDC XM, the sensitivity of Cross-Stat was 100%, the specificity was 93%, the positive predictive value was 73%, and the negative predictive value was 100%. A technical shortcoming of the Cross-Stat assay is that the frequency of indeterminate samples in the assays was 15%. Among 49 Cross-Stat negative vs. 13 Cross-Stat positive primary cadaveric renal allograft recipients (all AHG-CDC IgG-XM negative), there was no statistical difference in overall graft survival.
Conclusion:
Given the important theoretical advantages of enzyme-linked immunosorbent assay-based XM methods over the CDC XM, however, further testing of the clinical relevance of the Cross-Stat is warranted.
More Related Videos
07:25In Vitro Methods for Comparing Target Binding and CDC Induction Between Therapeutic Antibodies: Applications in Biosimilarity Analysis
Published on: May 4, 2017
06:15Characterization of Thymus-dependent and Thymus-independent Immunoglobulin Isotype Responses in Mice Using Enzyme-linked Immunosorbent Assay
Published on: September 7, 2018
Related Concept Videos
00:42Cross-reactivity
01:33Enzyme-Linked Immunosorbent Assay
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen. Enzyme-substrate reaction allows the antigen to be visualized or quantified.
01:22Immunocytochemistry and Immunohistochemistry
These...