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Kim, Lee, Hong, and Chai: Comparison of Two Enzyme-Linked Immunosorbent Assays for Detecting Parasitic Diseases
Original Article

Korean J Clin Microbiol 2008;11(1):56-62.

Published online: 20 April 2008

DOI: https://doi.org/10.5145/kjcm.2008.11.1.56

Comparison of Two Enzyme-Linked Immunosorbent Assays for Detecting Parasitic Diseases

Hye Ryoun Kim1, Mi Kyung Lee1, , Sung Tae Hong2, Jong Yil Chai2

1Department of Laboratory Medicine, Chung-Ang University College of Medicine, Seoul, Korea

2Department of Parasitology and Tropical Medicine, Seoul National University, Seoul, Korea

Correspondence: Mi Kyung Lee, Department of Laboratory Medicine, Chung-Ang University College of Medicine, 65-207, Hangangno 3-ga, Yongsan-gu, Seoul 140-757, Korea. (Tel) 82-2-748-9837, 9694, (Fax) 82-2-797-3471, (E-mail) cpworld@cau.ac.kr

Received 4 January 2008       Accepted 15 March 2008

Copyright © 2008 Korean Journal of Clinical Microbiology

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

Serologic tests for specific antibody nowadays are widely employed for the diagnosis of parasitic diseases. Recently, an increasing numbers of kits have adopted enzyme-linked immunosorbent assay (ELISA) for the detection of parasitic antibodies. In this study, we evaluated two ELISA reagents for the diagnosis of parasitic diseases.

Methods

A total of 553 serum and 156 CSF samples were assayed using an in-house micro-ELISA and GenediaⓇ Ab ELISA (Green cross PBM, Korea) for Cysticercus, Paragonimus westermani, Clonorchis sinensis, and Sparganum. We reviewed the medical records of all patients. The results from GenediaⓇ Ab ELISA kit were compared with those from the in-house micro-ELISA method.

Results

The overall concordance rate between the two ELISA tests was 95.5%. When compared with the clinical information, the sensitivity, specificity, positive predictive value, and negative predictive value of the in-house micro-ELISA were 100%, 99.0∼99.6%, 82.4∼96.4%, and 100%, and the respective figures for GenediaⓇ Ab ELISA kit were 92.9∼100%, 88.0∼ 97.3%, 41.7∼50%, and 99.9∼100% with kappa agreement of 0.53–0.63. Comparison of two ELISA methods showed a significant difference (P＜0.05). Retesting of 85 discordant samples showed that the concordance rate of the in-house ELISA was 97.7% and that of GenediaⓇ Ab ELISA was 28.2%.

Conclusion

GenediaⓇ Ab ELISA kit showed an intermediate level of kappa agreement compared with the in-house ELISA. Further studies are necessary to improve the concordance rate of the two methods, and a careful interpretation of these results is required for a precise diagnosis.

Keywords: Enzyme-Linked Immunosorbent Assay (ELISA), Cysticercus, Paragonimus west-ermani, Clonorchis sinensis, Sparganum

References

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Fig. 1.
Correlation between Optical Density values with in-house and Genedia ELISA.
kjcm-11-56f1.tif
Table 1.
Procedures and conditions of 2 different ELISAs employed in this study
  In-house ELISA Genedia-ELISA
Coating condition Serum dilution in PBS 4oC, overnight 4oC, overnight
1：100 1：20
   
Reaction condition Conjugate 37oC, 2 hours 37oC, 1 hour
HRP-antihuman HRP-Protein A
IgG  
Dilution in PBS Reaction condition Chromogen 1：8000 1：101
37oC, 2 hours 37oC, 1 hour
OPD in distilled Tetramethylbenzidine
water  
Reaction condition 37oC, Room temperature,
18 minutes 30 minutes
Reading absorbance 490 nm 450 nm

Abbreviations: ELISA, enzyme linked immunosorbent assay; HRP, horseradish peroxidase; OPD, o-phenylenediamine.

Table 2.
Comparison of in-house ELISA and Genedia ELISA by kappa agreement and McNemar test
In-house ELISA Paragonimus wertermani Cysticercus Sparganum Clonorchis sinensis
  Positive Negative Positive Negative Positive Negative Positive Negative
Genedia Positive 13 18 25 35 27 36 22 22
ELISA Negative 4 674 2 647 1 645 2 507
No. 709 709 709 553
Exact Sig.(2-tailed) P＜0.05 P＜0.05 P＜0.05 P＜0.05
Kappa agreement 0.527 0.551 0.570 0.626

Abbreviations: ELISA, enzyme linked immunosorbent assay.

Table 3.
Clinical information on the discrepancy between in-house ELISA and Genedia ELISA
Results Parasites (No. of case) Tx for parasites Clinical information (No. of cases)
in-house ELISA Genedia ELISA
Positive Negative PW (4) (−) Eosinophilia (1), fever (1), indetermined (1)
    (＋) Parasitemia (1)  
    Cysticercus (2) (−) Cross reaction (1), no clinical sx (1)
    Sparganum (1) (−) Liver disease (1)
    CS (2) (−) Eosinophilia (2), cross reaction (1)
Negative Positive PW (18) (−) Eosinophilia (9), pain (3), headache (2), no clinical sx (3), fever (1)
    Cysticercus (35) (−) Eosinophilia (16), pain (2), headache (6), no clinical sx (9), fever (1), indetermined (1)
    Sparganus (36) (−) Eosinophilia (18), cross reaction (3), pain (4), headache (3), no clinical sx (7), indetermined (1)
    CS (22) (−) Eosinophilia (12), cross reaction (2), pain (4), no clinical sx (2), fever (2)

Abbreviations: ELISA, enzyme linked immunosorbent assay; PW, Paragonimus westermani; CS, Clonorchis sinensis; sx, symptom; Tx, treatment.

Table 4.
Diagnostic performances (sensitivity, specificity, positive and negative predictive values) of Genedia ELISA and in-house ELISA
No. of case Cysticercus (%) PW (%) Sparganum (%) CS (%)
327 327 327 261
In-house Genedia In-house Genedia In-house Genedia In-house Genedia
ELISA ELISA ELISA ELISA ELISA ELISA ELISA ELISA
Sensitivity 100 100 100 92.9 100 100 100 100
(25/25) (25/25) (14/14) (13/14) (27/27) (27/27) (22/22) (22/22)
Specificity 99.3 88.4 99.0 94.5 99.7 88.0 99.1 90.7
(300/302) (267/302) (310/313) (295/313) (299/300) (264/300) (237/239) (217/239)
PPV 92.6 41.7 82.4 41.9 96.4 42.9 91.7 50
(25/27) (25/60) (14/17) (13/31) (27/28) (27/63) (22/24) (22/44)
NPV 100 100 100 99.7 100 100 100 100
(300/300) (267/267) (310/310) (295/296) (299/299) (264/264) (237/237) (217/217)

Abbreviations: PPV, positive predictive value; NPV, negative predictive value; PW, Paragonimus westermani; CS, Clonorchis sinensis.

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