kjlm The Korean Journal of Laboratory Medicine Korean J Lab Med 2383-5702 2383-5710 The Korean Society for Laboratory Medicine 10.3343/kjlm.2007.27.6.437 kjlm-27-437 Original Article Comparisons of Latex Agglutination, Immunochromatography and Enzyme Immunoassay Methods for the Detection of Rotavirus Antigen LeeSook YoungM.D. 1 HongJong HeeM.T. 1 LeeSun WhaM.D. 2 LeeMiaeM.D. 1 Department of Laboratory Medicine, Ewha Womans University, School of Medicine, Seoul, Korea Neodin Medical Institute, Seoul, Korea 022007 14022007 276 437441 17092007 01112007 01112007 Copyright © 2007 by the Korean Society for Laboratory Medicine 2007 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. 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.

 Rotavirus Latex agglutination Immunochromatography Enzyme Immunoassay REFERENCES Wilhelmi I Roman E Sanchez-Fauquier A Viruses causing gastroenteritis Clin Microbiol Infect 20039 24762 Anderson EJ Weber SG Rotavirus infection in adults Lancet Infect Dis 20044 919 Kim HS Jang MW Kim CH Lee HH Yoo HJ Clinical study of rotavirus gastroenteritis in the last 2 years Korean J Pediatr 200548 111620 (김형식, 장명완, 김철홍, 이현희, 유황재. 최근 2년간로타바이러스장염에대한임상적연구. 소아과 2005;48: 1116-20). Knisley CV Bednarz-Prashad AJ Pickering LK Detection of rotavirus in stool specimens with monoclonal and polyclonal antibody-based assay systems J Clin Microbiol 198623 897900 Dennehy PH Schutzbank TE Thorne GM Evaluation of an automated immunodiagnostic assay, VIDAS Rotavirus, for detection of rotavirus in fecal specimens J Clin Microbiol 199432 8257 Dennehy PH Gauntlett DR Spangenberger SE Choice of reference assay for the detection of rotavirus in fecal specimens: electron microscopy versus enzyme immunoassay J Clin Microbiol 199028 12803 Raboni SM Noguiera MB Hakim VM Torrecilha VT Lerner H Tsuchiya LR Comparison of latex agglutination with enzyme immunoassay for detection of rotavirus in fecal specimens Am J Clin Pathol 2002117 3924 Sambourg M Goudeau A Courant C Pinon G Denis F Direct appraisal of latex agglutination testing, a convenient alternative to enzyme immunoassay for the detection of rotavirus in childhood gastroenteritis, by comparison of two enzyme immunoassays and two latex tests J Clin Microbiol 198521 6225 Pai CH Shahrabadi MS Ince B Rapid diagnosis of rotavirus gastroenteritis by a commercial latex agglutination test J Clin Microbiol 198522 84650 Brandt CD Arndt CW Evans GL Kim HW Stallings EP Rodriguez WJ Evaluation of a latex test for rotavirus detection J Clin Microbiol 198725 18002 Wilhelmi I Colomina J Martin-Rodrigo D Roman E Sanchez-Fauquier A New immunochromatographic method for rapid detection of rotaviruses in stool samples compared with standard enzyme immunoassay and latex agglutination techniques Eur J Clin Microbiol Infect Dis 200120 7413 Dennehy PH Hartin M Nelson SM Reising SF Evaluation of the ImmunoCardSTAT! rotavirus assay for detection of group A rotavirus in fecal specimens J Clin Microbiol 199937 19779 Persing DH Smith TF Diagnostic molecular microbiology: principles and applications 1st ed. Washinton D.C. American Society for Microbiology 19933838 Dennehy PH Gauntlett DR Tente WE Comparison of nine commercial immunoassay for the detection of rotavirus in fecal specimens J Clin Microbiol 198826 16304 Kang JO Kim SE Kim TY Park IK Choi TY Trends in rotavirus gastroenteritis in Korea from 1989 through 1998 and comparison of Slidex Rota-kit 2 and VIDAS Rotavirus Korean J Clin Microbiol 19992 1527 (강정옥, 김선의, 김신규, 박일규, 최태열. 로타바이러스성장염의유행경향변화및 Slidex Rota-Kit 2와 VIDAS Rotavirus의비교. a대한임상미생물학회지 1999;2: 152-7.) Eing BR May G Baumeister HG Kuhn JE Evaluation of two enzyme immunoassays for detection of human rotaviruses in fecal specimens J Clin Microbiol 200139 45324 Tables

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.

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.

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.

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.