Article information
Abstract
Background and objectives
Rotavirus (RV) is the main etiological agent of diarrhea in childhood; its laboratory diagnosis is crucial to guide the clinical management and prevention of its spread. RV immunization was introduced in Brazilian 6-month-old children in 2006. The present study was aimed to evaluate three methodologies used for human RV detection in stool samples obtained from patients hospitalized due to gastroenteritis in a teaching hospital and report the impact of RV immunization in hospitalization by diarrhea.
Methods293 stool samples collected in the 2001-2008 period were analyzed by enzyme immunoassay (EIA), latex agglutination (LA) and polyacrylamide gel electrophoresis (PAGE).
ResultsRotavirus was detected in 34.8% of samples by LA assay, 28.3% of samples by EIA assay and in 25.6% of samples by PAGE assay. Considering the PAGE method as gold standard, the sensitivity, specificity and accuracy of EIA were 94.6%, 94.4% and 94.5%, and to LA were 82.6%, 81.6% and 81.9%, respectively.
ConclusionsThese results indicate that antigen detection by EIA is a rapid, sensitive and specific method, and could be used in large-scale applications for screening stool samples suspected of RV infection. This study showed decreased incidence of RV infection in hospitalized children prior to the implementation of the national immunization program against RV.
Keywords:
rotavirus
diarrhea
immunoenzyme techniques
latex fixation tests
electrophoresis
polyacrylamide gel
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References
[1]
S.G. Morillo, A. Luchs, A. Cilli, F.F. Costa, R.C. Carmona, M.C. Timenetsky.
Characterization of rotavirus strains from day care centers: pre- and post-rotavirus vaccine era.
J Pediatr. (Rio J), 86 (2010), pp. 155-158
[2]
U.D. Parashar, C.J. Gibson, J.S. Bresse, R.I. Glass.
Rotavirus and severe childhood diarrhea.
Emerg Infect Dis., 12 (2006), pp. 304-306
[3]
A. Kapikian, Y. Hoshino, R. Chanock.
Rotaviruses.
Fields Virology,
[4]
A.C. Linhares.
Rotavirus Infection in Brazil: Epidemiology.
Immunity, and Potential Vaccination. Braz J Infect Dis., 1 (1997), pp. 284-293
[5]
J.D. Mascarenhas, R.H. Gusmao, C.R. Barardi, et al.
Characterization of rotavirus P genotypes circulating among paediatric inpatients in northern Brazil.
Rev Inst Med Trop Sao Paulo, 41 (1999), pp. 165-170
[6]
A. Meneghetti, A. Bolognini, F. Lauretti, R. Linhares, N. Santos, C. Nozawa.
Incidence of group A rotavirus in urban and rural areas of the city of Londrina - Brazil, from 1995 to 1997.
Braz Arch Biol Technol., 44 (2001), pp. 257-261
[7]
V. Richardson, J. Hernandez-Pichardo, M. Quintanar-Solares, et al.
Effect of rotavirus vaccination on death from childhood diarrhea in Mexico.
N Engl J Med., 362 (2010), pp. 299-305
[8]
A.M. Sartori, J. Valentim, P.C. de Soarez, H.M. Novaes.
Rotavirus morbidity and mortality in children in Brazil.
Rev Panam Salud Publica, 23 (2008), pp. 92-100
[9]
M.B. Souza, M.L. Racz, J.P. Leite, C.M. Soares, R.M. Martins, V. Munford, D.D. Cardoso.
Molecular and serological characterization of group a rotavirus isolates obtained from hospitalized children in Goiania.
Brazil, 1998-2000. Eur J Clin Microbiol Infect Dis., 22 (2003), pp. 441-443
[10]
M.C. Timenetsky, N. Santos, V. Gouvea.
Survey of rotavirus G and P types associated with human gastroenteritis in Sao Paulo.
Brazil, from 1986 to 1992. J Clin Microbiol., 32 (1994), pp. 2622-2624
[11]
R.F. Bishop, G.P. Davidson, I.H. Holmes, B.J. Ruck.
Virus particles in epithelial cells of duodenal mucosa from children with acute non-bacterial gastroenteritis.
Lancet, 2 (1993), pp. 1281-1283
[12]
U. Desselberger.
Molecular epidemiology of rotaviruses.
Viruses and the gut,
[13]
M. Estes.
Rotaviruses and their replication.
Fields Virology, 4 ed,
[14]
M.K. Estes, J. Cohen.
Rotavirus gene structure and function.
Microbiol Rev, 53 (1989), pp. 410-419
[15]
R. Cauás, A. Falbo, J. Correia, K. Oliveira, F. Montenegro.
Diarréia por rotavírus em crianças desnutridas hospitalizadas no Instituto Materno Infantil Prof Fernando Figueira IMIP.
Rev Bras Saude Mater Infant, 6 (2006), pp. s77-s83
[16]
P. Costa, S. Grisi, D. Cardoso, F. Fiaccadori, M. Souza, R. Santos.
Clinical and epidemiological trends in rotavirus A infections.
Pediatria (São Paulo), 26 (2004), pp. 151-158
[17]
A. Nunes, L. Mello, R. Parrode, J. Bittar.
Prevalence of rotavirus in feces of children with acute diarrhea: clinical signs and associated symptoms.
Revista da AMRIGS, 54 (2010), pp. 147-151
[18]
G.M. Beards, A.D. Campbell, N.R. Cottrell, et al.
Enzyme-linked immunosorbent assays based on polyclonal and monoclonal antibodies for rotavirus detection.
J Clin Microbiol, 19 (1984), pp. 248-254
[19]
J. Buesa, J. Colomina, J. Raga, A. Villanueva, J. Prat.
Evaluation of reverse transcription and polymerase chain reaction (RT/PCR) for the detection of rotaviruses: applications of the assay.
Res Virol, 147 (1996), pp. 353-361
[20]
T. Ferreira, M. Becho, A. Bernardo, et al.
Performance of a latex agglutination test in the diagnosis of acute gastroenteritis by rotavirus.
Braz J Microbiol, 37 (2006), pp. 587-589
[21]
S. Kasempimolporn, S. Louisirirotchanakul, P. Sinarachatanant, C. Wasi.
Polyacrylamide gel electrophoresis and silver staining for detection of rotavirus in stools from diarrheic patients in Thailand.
J Clin Microbiol, 26 (1988), pp. 158-160
[22]
G.M. Ruiz-Palacios, I. Perez-Schael, F.R. Velazquez, et al.
Safety and efficacy of an attenuated vaccine against severe rotavirus gastroenteritis.
N Engl J Med, 354 (2006), pp. 11-22
[23]
R. Boom, C.J. Sol, M.M. Salimans, C.L. Jansen, P.M. Wertheim-van Dillen, J. van der Noordaa.
Rapid and simple method for purification of nucleic acids.
J Clin Microbiol, 28 (1990), pp. 495-503
[24]
U.K. Laemmli.
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature, 227 (1970), pp. 680-685
[25]
A.J. Herring, N.F. Inglis, C.K. Ojeh, D.R. Snodgrass, J.D. Menzies.
Rapid diagnosis of rotavirus infection by direct detection of viral nucleic acid in silver-stained polyacrylamide gels.
J Clin Microbiol, 16 (1982), pp. 473-477
[26]
K. Bellamy, P.S. Gardner, M.H. Hambling, S. Rice, A.F. Bradburne.
Enzyme-linked immunosorbent assay for the detection of human rotavirus in stools.
J Virol Methods, 7 (1983), pp. 65-72
[27]
L. Berthiaume, R. Alain, B. McLaughlin, P. Payment, P. Trepanier.
Rapid detection of human viruses in faeces by a simple and routine immune electron microscopy technique.
J Gen Virol, 55 (1981), pp. 223-227
[28]
P.C. Grauballe, Z. Jarzabek.
Comparison of indirect double antibody and double antibody sandwich ELISA techniques with latex agglutination test for the diagnosis of human rotavirus infection.
Acta Virol, 28 (1984), pp. 59-63
[29]
P.J. Middleton, M.D. Holdaway, M. Petric, M.T. Szymanski, J.S. Tam.
Solid-phase radioimmunoassay for the detection of rotavirus.
Infect Immun, 16 (1977), pp. 439-444
[30]
O. Nakagomi, A. Nakagomi, T. Suto, et al.
Detection of human rotavirus by reversed passive hemagglutination (RPHA) using antibody against a cultivable human rotavirus as compared with electron microscopy (EM) and enzyme-linked immunosorbent assay (ELISA).
Microbiol Immunol, 26 (1982), pp. 747-751
[31]
S.M. Raboni, M.B. Nogueira, V.M. Hakim, V.T. Torrecilha, H. Lerner, L.R. Tsuchiya.
Comparison of latex agglutination with enzyme immunoassay for detection of rotavirus in fecal specimens.
Am J Clin Pathol, 117 (2002), pp. 392-394
[32]
E. Carraro, A. Perosa, I. Siqueira, J. Pasternak, M. Martino.
Four kits evaluation for rotavirus rapid antigen detection in stool samples.
Rev Bras Anal Clin, 40 (2008), pp. 309-311
[33]
P.J. Masendycz, E.A. Palombo, R.J. Gorrell, R.F. Bishop.
Comparison of enzyme immunoassay PCR, and type-specific cDNA probe techniques for identification of group A rotavirus gene 4 types (P types).
J Clin Microbiol, 35 (1997), pp. 3104-3108
[34]
A. Momenzadeh, S. Modarres, A. Faraji, et al.
Comparision of enzyme immunoassay, immunochromatography, and RNA-polyacrylamide gel electrophoresis for diagnosis of rotavirus infection in children with acute gastroenteritis.
Iran J Med Sci, 33 (2008), pp. 173-176
[35]
A.C. Linhares.
Rotavirus infection in Brazil: epidemiology and challenges for its control.
Cad Saude Publica, 16 (2000), pp. 629-646
[36]
V. Munford, E. Souza, T. Caruzo, M. Martinez, M. Rácz.
Serological and molecular diversity of human rotavirus in São Paulo.
Brazil. Braz J Microbiol, 38 (2007), pp. 459-466
[37]
J. Bresee, Z.Y. Fang, B. Wang, et al.
First report from the Asian Rotavirus Surveillance Network.
Emerg Infect Dis, 10 (2004), pp. 988-995
[38]
U.D. Parashar, E.G. Hummelman, J.S. Bresee, M.A. Miller, R.I. Glass.
Global illness and deaths caused by rotavirus disease in children.
Emerg Infect Dis, 9 (2003), pp. 565-572
[39]
M. Altindis, S. Yavru, A. Simsek, A. Ozkul, A. Ceri, H. Koc.
Rotavirus infection in children with acute diarrhea as detected by latex agglutination ELISA and polyacrylamide gel electrophoresis.
Indian Pediatr, 41 (2004), pp. 590-594
[40]
J.A. Gomez, J.A. Bercovich, S. Grinstein.
Comparison of enzyme immunoassay, counterimmunoelectrophoresis and polyacrylamide gel electrophoresis for the diagnosis of rotaviruses.
Rev Argent Microbiol, 17 (1985), pp. 111-114
[41]
O.S. Ibrahim, D. Sunderland, C.A. Hart.
Comparison of four methods for detection of rotavirus in faeces.
Trop Doct, 20 (1990), pp. 30-32
[42]
A. Chakravarti, S. Kumar, S.K. Mittal, S. Broor.
Comparison of latex agglutination and polyacrylamide gel electrophoresis with enzyme linked immunosorbent assay for detecting human rotavirus in stool specimens.
Indian Pediatr, 28 (1991), pp. 507-510
[43]
C.D. Brandt, C.W. Arndt, G.L. Evans, et al.
Evaluation of a latex test for rotavirus detection.
J Clin Microbiol, 25 (1987), pp. 1800-1802
[44]
E. Cornaglia, M. Barrandeguy, N. Fijtman, A. Schudel.
Enzyme linked immunosorbent, immunofluorescence test and electrophoretic analysis of rotaviral RNA in the diagnosis and characterization of the bovine rotavirus.
Rev Latinoam Microbiol, 31 (1989), pp. 59-62
[45]
A. Carducci, R. Vannucchi, M. Guidi, D. Reali, M.A. Ruschi.
Human rotavirus detection in stool specimens using enzyme-linked immunosorbent assays and latex agglutination test.
Boll Ist Sieroter Milan, 67 (1988), pp. 241-244
[46]
P.H. Dennehy, D.R. Gauntlett, W.E. Tente.
Comparison of nine commercial immunoassays for the detection of rotavirus in fecal specimens.
J Clin Microbiol, 26 (1988), pp. 1630-1634
[47]
E.E. Thomas, M.L. Puterman, E. Kawano, M. Curran.
Evaluation of seven immunoassays for detection of rotavirus in pediatric stool samples.
J Clin Microbiol, 26 (1988), pp. 1189-1193
[48]
A. Pirkooh, M. Shahrabadi.
Development of a latex agglutination method for diagnosis of rotavirus infections.
Iran J Med Sci, 32 (2007), pp. 100-104
[49]
R.K. Maes, D.L. Grooms, A.G. Wise, et al.
Evaluation of a human group a rotavirus assay for on-site detection of bovine rotavirus.
J Clin Microbiol, 41 (2003), pp. 290-294
[50]
A. Jensen.
Evaluation of two commercial kits for rapid detection of human rotavirus in feces: Rotalex, a latex agglutination test and Rotavirus ELISA Kit.
Acta Pathol Microbiol Immunol Scand B, 93 (1985), pp. 159-160
[51]
C. Rodrigues, J. Zattar, A. Rossoni, E. Stroparo, D. Roque, L. Romagnoli.
Impact of vaccination against rotavirus in outpatients visitis and hospitalization due to gastroenteritis in a university hospital in Brazil 6th Annual Meeting of the World Society for pediatric Infectious Diseases.
Buenos Aires, (2009),
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