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Lee, Hong, Lee, and Lee: Comparisons of Latex Agglutination, Immunochromatography and Enzyme Immunoassay Methods for the Detection of Rotavirus Antigen
Original Article

Korean J Leg Med 2007;27(6):437-441.

Published online: 14 February 2007

DOI: https://doi.org/10.3343/kjlm.2007.27.6.437

Comparisons of Latex Agglutination, Immunochromatography and Enzyme Immunoassay Methods for the Detection of Rotavirus Antigen

Sook Young Lee, M.D.1, Jong Hee Hong, M.T.1, Sun Wha Lee, M.D.2, Miae Lee, M.D.1

1Department of Laboratory Medicine, Ewha Womans University, School of Medicine, Seoul, Korea

2Neodin Medical Institute, Seoul, Korea

Received 17 September 20071 November 2007       Accepted 1 November 2007

Copyright © 2007 by the 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

Rotavirus is the most common cause of childhood gastroenteritis during winter season. Rapid, accurate diagnosis is essential for preventing severe complications of rotaviral gastroenteritis. The sensitivity and specificity of five detection test kits for rotavirus including latex agglutination (LAT), enzyme immunoassay (EIA) and three immunochromatographic methods (ICG) were evaluated in this study.

Methods

A total of 95 stool samples collected from patients with acute gastroenteritis were studied. The test kits were as follows: LAT (Slidex latex, bioMerieux Vitek, France); three kinds of ICG (Dipstick ROTA, Eiken, Japan; SAS Rota Test, SA Scientific, Inc., USA; and ASAN Easy Test Rota strip, ASAN Pharmaceutical., Korea); and EIA (VIDAS Rotavirus, bioMerieux Vitek). The samples showing discordant results were reevaluated by reverse-transcription (RT) PCR and clinical manifestations.

Results

Of a total of 95 cases, 56 (58.9%) were positive and 39 (41.1%) were negative. Thirteen cases showed discordant results. Sensitivity and specificity were, respectively, 85.7% and 100% for LAT, 100% and 95% for both of Dipstick ROTA and SAS Rota, 86.7% and 87.5% for ASAN Rota strip and 98.1% and 97.3% for EIA.

Conclusions

LAT was rapid and easy to perform and showed the lowest sensitivity among the five test kits. ICG showed a good agreement with EIA and RT-PCR. EIA was the best in respect of sensitivity and specificity, but difficulty in interpretations of equivocal results and time-consuming procedures were limitations. In conclusion, ICG, which is easy to perform at a low cost, may be an optimal method in place of LAT for the detection of rotavirus.

Keywords: Rotavirus, Latex agglutination, Immunochromatography, Enzyme Immunoassay

REFERENCES

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Table 1.
Characteristics of the five different tests for the detection of rotavirus
Methods Slidex Rota-Kit2 Dipstick ROTA SAS Rota Test ASAN Easy Test Rota strip VIDAS Rotavirus
Principle Target antigen LAT Anti-rotavirus monoclonal Ab ICG (dipstick) Capsid protein ICG (device) Capsid protein ICG (strip) VP6 ELFA VP6
Sample preparation Stool 1-2 mL or supernatant after centrifuge Stool 12.5 μL or supernatant after centrifuge Stool 30-50 mL or supernatant after centrifuge Stool 12.5 μL or supernatant after centrifuge Stool 0.5 mL or supernatant after centrifuge
Analysis time 2 min 15 min 15 min 15 min 30 min
Positive result Agglutination Colored lines on specimen and control Colored lines on specimen and control Colored lines on specimen and control Automated analysis of fluorescence on 450 nm

Abbreviations: LAT, Latex agglutination; ICG, Immunochromatography; ELFA, Enzyme linked fluorescent assay.

Table 2.
Concordance rate of five different assays for diagnosis of rotavirus
LAT Dipstick ROTA SAS Rota ASAN Rota strip EIA N (%)
- - - - - 35 (36.8)
+ + + + + 47 (49.4)
- - - - Equivocal 2 (2.1)
- + + + - 1 (1.1)
- + + + + 4 (4.2)
- + + + Equivocal 2 (2.1)
+ + + - + 1 (1.1)
- + + - - 1 (1.1)
- + + - Equivocal 1 (1.1)
- + + - + 1 (1.1)

Abbreviations: LAT, Latex agglutination; EIA, Enzyme Immunoassay.

Table 3.
RT-PCR results and clinical review of 13 discordant cases
No. Case LAT Dipstick ROTA SAS Rota ASAN Rota strip EIA (test value)∗ RT-PCR Clinical diagnosis Final interpretation
67 - - - NT Eq (104) - AGE, Imperfecta anus -
42 - - - - Eq (165) - GERD -
37 - + + + - (1) - AGE -
16 - + + + + (2754) - Neonatal convulsion -
11 - + + + + (1377) + Rotaviral enteritis +
17 + + + - + (1836) NT Rotaviral enteritis +
27 - + + - - (30) + Rotaviral enteritis +
28 - + + + Eq (255) + TTN +
29 - + + + + (2355) + TTN +
43 - + + - + (529) + AGE +
48 - + + + Eq (108) + AGE +
50 - + + - Eq (184) + Rotaviral enteritis +
52 - + + + + (438) + Convulsion +

∗ EIA test value, patient's test Relative Fluorescence Value (RFV)-Standard RFV.

Abbreviations: RT-PCR, Reverse-transcription PCR; LAT, Latex agglutination; EIA, Enzyme Immunoassay; AGE, Acute Gastroenteritis; GERD, Gastroe-sophageal reflux disease; TTN, Transient tachypnea of newborn; Eq, equivocal; NT, not tested.

Table 4.
Sensitivity and specificity of five different assays after resolution of discordant cases by reverse-transcription PCR
Parameter Test methods (N)
LAT (95) Dipstick ROTA (95) SAS Rota (95) ASAN Rota strip (46) EIA (90)∗
True positive 48 56 56 26 52
True negative 39 37 37 14 36
False positive (%) 0 (0.0) 2 (2.1) 2 (2.1) 2 (4.3) 1 (1.1)
False negative (%) 8 (8.4) 0 (0.0) 0 (0.0) 4 (8.6) 1 (1.1)
Sensitivity (%) 85.7 100.0 100.0 86.7 98.1
Specificity (%) 100.0 95.0 95.0 87.5 97.3

Final positive cases 56, negative cases 39.

∗ Excluding five equivocal results.

Abbreviations: LAT, Latex agglutination; ICG, Immunochromatography; EIA, Enzyme Immunoassay.

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