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Jung, Oh, Yang, Ahn, Kim, Park, and Han: Comparative Evaluation of ELISA and Luminex Panel Reactive Antibody Assays for HLA Alloantibody Screening
Original Article

Korean J Leg Med 2009;29(5):473-480.

Published online: 14 January 2009

DOI: https://doi.org/10.3343/kjlm.2009.29.5.473

Comparative Evaluation of ELISA and Luminex Panel Reactive Antibody Assays for HLA Alloantibody Screening

Seonkyung Jung, M.D.1, Eun-Jee Oh, M.D.1, Chul-Woo Yang, M.D.2, Woong-Shick Ahn, M.D.3, Yonggoo Kim, M.D.1, Yeon-Joon Park, M.D.1, Kyungja Han, M.D.1

1Departments of Laboratory Medicine, Seoul, Korea

2Internal Medicine, and Obstetrics and Gynecology, Seoul, Korea

3The Catholic University of Korea, Seoul, Korea

Corresponding author: Eun-Jee Oh, M.D. Department of Laboratory Medicine, Seoul St. Mary's Hospital, 505 Banpo-dong, Seocho-gu, Seoul 137-701, Korea Tel: +82-2-2258-1641, Fax: +82-2-2258-1719 E-mail: ejoh@catholic.ac.kr

∗ This study was supported by the Catholic Medical Center Research Foundation made in the program year of 2007.

      Revised 20 July 200924 September 2009       Accepted 30 September 2009

Copyright © 2009 Korean Society for Legal Medicine

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background:

For the detection of HLA antibodies, solid-phase tests using purified HLA antigens are increasingly used. In this study, we analyzed the panel reactive antibody (PRA) test results using ELISA and Luminex methods, and the results were compared with those of crossmatch test.

Methods:

A total of 111 sera including 90 sera from kidney transplanted patients were tested. ELISA-PRA was performed using Lambda Antigen Tray Class I and II Mixed kits (One Lambda Inc., USA) and additional test was performed to identify HLA specificities. Luminex-PRA tests were performed using LABScreen Mixed kits (One Lambda Inc., USA) and LIFECODES LifeScreen Deluxe kits (Tepnel Co., USA).

Results:

The positive rates of PRA were higher in Tepnel (P=0.006) and One Lambda Luminex (P<0.001) methods than ELISA, without significant difference between two Luminex methods (P= 0.087). The overall concordance rate among the three PRA tests was 62.2% (69/111). The positive and negative predictive values of PRA tests for the flow cytometric crossmatch were 33.3-45.7% and 85.7-89.5%, respectively. Of the two Luminex methods, One Lambda showed higher positive rate than Tepnel for the detection of class I antibodies. The sensitivity of pretransplant PRA for the detection of posttransplant acute rejection episodes was higher in Luminex (P=0.007 for Tepnel, P=0.003 for One lambda) than ELISA method.

Conclusions:

Different methods used to detect HLA antibodies showed discrepant results. As the Luminex method was more sensitive than ELISA for the detection of HLA antibodies, it can be used as a routine test in the transplantation laboratory.

Keywords: Panel reactive antibodies, HLA antibodies, Luminex-PRA, ELISA-PRA

REFERENCES

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Table 1.
Positive rate of panel reactive antibody tests in 111 sera
Antibody specificities % (N of positive cases)
ELISA Tepnel-Luminex One Lambda-Luminex
Class I and/or II 59.5% (66) 76.6% (85) 85.6% (95)
Class I 52.3% (58) 64.9% (72) 84.7% (94)
Class II 31.5% (35) 62.2% (69) 55.9% (62)

∗ For Class I and/or II, ELISA vs Tepnel, P=0.006; ELISA vs One Lambda, P<0.001; Tepnel vs One Lambda, P=0.087. For Class I, ELISA vs Tepnel, P=0.057; ELISA vs One Lambda, P<0.001; Tepnel vs One Lambda, P=0.001. For Class II, ELISA vs Tepnel, P<0.001, ELISA vs One Lambda, P<0.001.

Table 2.
Comparison between crossmatch and PRA screening results in 68 sera
Crossmatch Positive N (%)
ELISA-PRA Tepnel-Luminex-PRA One Lambda-Luminex-PRA
CDCXM(+)/FCXM(+) (N=12) 12 (100) 12 (100) 11 (91.7)
CDCXM(+)/FCXM(-) (N=1) 0 (0) 0 (0) 1 (100)
CDCXM(-)/FCXM(+) (N=8) 4 (50.0) 6 (75.0) 7 (87.5)
CDCXM(-)/FCXM(-) (N=47) 19 (40.4) 31 (66.0) 35 (74.5)

Abbreviations: CDCXM, complement dependent cytotoxicity crossmatch; FCXM, flowcytometric crossmatch; +, positive; -, negative.

Table 3.
Concordance rates of the results of three PRA methods in 111 sera
Antibody specificities ELISA, Tepnel-Luminex, and One Lambda-Luminex ELISA and Tepnel-Luminex ELISA and One Lambda-Luminex Tepnel-Luminex and One Lambda-Luminex
Class I and/or II 62.2% 77.5% 68.5% 77.5%
Class I 55.9% 78.4% 62.2% 71.2%
Class II 60.4% 64.0% 72.1% 84.7%

For class I, ‘ELISA and Tepnel’ vs ‘ELISA and One Lambda’, P=0.008.

For class II, ‘ELISA and Tepnel’ vs ‘ELISA and One Lambda’, P<0.001; ‘ELISA and One Lambda’, vs ‘Tepnel and One Lambda’, P=0.023.

Table 4.
Predictive value of PRA screening tests for CDC or flow cytometric crossmatch
PRA methods For CDCXM For FCXM
PPV NPV PPV NPV
ELISA 34.3% 97.0% 45.7% 87.9%
  (12/35) (32/33) (16/35) (29/33)
Tepnel-Luminex 24.5% 94.7% 36.7% 89.5%
  (12/49) (18/19) (18/49) (17/19)
One Lambda-Luminex 22.2% 92.9% 33.3% 85.7%
  (12/54) (13/14) (18/54) (12/14)

Abbreviations: CDCXM, complement dependent cytotoxicity crossmatch; FCXM, flowcytometric crossmatch; PPV, positive predictive value; NPV, negative predictive value.

Table 5.
Performance data of pre-transplant PRA screening tests for the detection of acute rejection episodes in 48 kidney transplanted patients
PRA methods Sensitivity Specificity PPV NPV
ELISA 31.8% 61.5% 41.2% 51.6%
Tepnel-Luminex 72.7% 38.5% 50.0% 62.5%
One Lambda-Luminex 77.3% 7.7% 41.5% 28.6%

Abbreviations: See Table 4.

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