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      Brote epidémico de meningitis viral causado por echovirus tipo 4 en la provincia de Misiones Translated title: Outbreak of viral meningitis caused by echovirus type 4 in Misiones province

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          Abstract

          Se realizó un estudio retrospectivo a fin de describir un brote epidémico de meningitis causado por enterovirus, que comprometió a 143 niños de 1 mes a 14 años internados en el Hospital Pediátrico de Posadas (Misiones) con diagnóstico de meningitis aséptica, entre agosto y diciembre de 2005. Se observó un aumento de casos entre las semanas 33 a 50, con un pico máximo entre las semanas 47 y 48, lo que confirmó el brote. La mediana de edad de los niños afectados fue de 8 años y el 55,2% fueron varones. El 80% de los casos se observó entre escolares (5 a 14 años). El promedio del tiempo de internación fue de 4,5 ± 1,7 días, y no se registraron fallecidos. Los LCR se estudiaron mediante examen citoquímico y estudios bacteriológicos y virológicos (aislamiento viral, RT- PCR anidada e identificación molecular mediante secuenciación génica). Los recuentos de células en LCR variaron entre 6 y 5040 células /mm3, el 92% fueron inferiores a 500 células/mm3 y el 43,5% mostró predominio linfocitario. El 56% presentó concentraciones de glucosa normal, con proteínas ligeramente elevadas. El 28% de las muestras estudiadas por cultivo (17/60) mostró efecto citopático, compatible con enterovirus. La RT-PCR anidada permitió detectar enterovirus en un 73% de las muestras (43/59), con 6 casos que se tipificaron como echovirus tipo 4. El índice de positividad al combinar ambas técnicas alcanzó el 83%.

          Translated abstract

          A descriptive retrospective study was carried out to describe an epidemic outbreak of enteroviral meningitis in Misiones. We reviewed records of 143 children from 1 month to 14 years of age who were hospitalized with aseptic meningitis in the Pediatric Hospital of Posadas from August to December 2005. Increased number of cases was observed between weeks 33 to 50 which reached a maximum peak in weeks 47 and 48, confirming an outbreak. The median of age was 8 years old, 55.2% were males. Eighty percent of cases were in 5 to 14 years old children. The average length of time spent in the hospital was 4.5±1.7 days, no deaths were reported. We performed cell counts, chemical and bacterial studies of CSF, and culture or RT-Nested/PCR for enteroviruses. Isolates were serotyped by RT-PCR amplification and genetic sequencing. Cell counts were from 6 to 5040 cells/mm3. Ninety two percent had less than 500 cells/mm3 and 43.5% had lymphocyte predominance. Glucose levels were normal with slightly elevated protein counts in 56% of cases. Of the cultured samples, 28% (17/60) showed cytopathic effect compatible with enterovirus. RT-n-PCR detected enterovirus in 73% (43/59) of the analyzed CSF. Echovirus type 4 was identified in 6 of them. The positive indicator obtained by combining both techniques was 83% (58/70).

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          Most cited references44

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          Enteroviral infections of the central nervous system.

          H Rotbart (1995)
          Infections of the CNS with the nonpolio enteroviruses are common and important causes of morbidity in both children and adults. Studies have recently defined the short-term and long-term outcomes of aseptic meningitis due to the enteroviruses. Focal encephalitis is increasingly recognized as a complication of enterovirus infection. Patients at greatest risk for sequelae of CNS enteroviral disease include neonates and those who are immunocompromised. The clinical presentation may mimic that of bacterial or other viral CNS infections, a circumstance making laboratory diagnosis of paramount importance for reducing unnecessary hospitalization and therapy. Recent advances in PCR technology, including its adaptation to a colorimetric microwell plate format, promise to greatly facilitate diagnosis of enteroviral infections. Promising antiviral drugs for CNS disease and other serious manifestations of enteroviral infections are under development.
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            New method for the extraction of viral RNA and DNA from cerebrospinal fluid for use in the polymerase chain reaction assay.

            A new, rapid, and simple method for the isolation of either RNA or DNA from cerebrospinal fluid samples for subsequent amplification by specific polymerase chain reaction (PCR) assays is described. The technique involves a single extraction with a guanidinium thiocyanate acid (GuSCN) buffer, and does not require the use of organic solvents. Applied to the recovery of enteroviral RNA, herpes simplex virus (HSV) and Varicella-zoster virus (VZV) DNAs the method has proved to be of equivalent or better efficiency than established methods of nucleic acid separation but is less laborious and time consuming. The simplicity of the procedure permits the processing of large numbers of samples and the use of a single preparative method for either RNA or DNA PCR makes it an attractive method for the routine laboratory.
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              Molecular characterization of human enteroviruses in clinical samples: comparison between VP2, VP1, and RNA polymerase regions using RT nested PCR assays and direct sequencing of products.

              Three nested RT-PCR assays were developed to permit sensitive typing of enteroviruses directly from clinical samples. These assays amplified short fragments from different genomic regions codifying for three proteins: VP2, VP1, and RNA polymerase. Given that enteroviruses have a high rate of degeneration within target codons among serotypes, the primers used consisted of mixed base and deoxyinosine residues. These techniques detected at 0.03-0.003 TCID50 of prototype Poliovirus 1 and Echovirus 30. They were used to characterize the enteroviral RNA detected in 18 CSF, stool, and throw swab samples and in 8 enterovirus isolates from patients with several syndromes. Phylogenetic analysis in each independent sequenced region grouped the enterovirus into four clusters, enabling genetic classification. A comparative study was performed among the 26 sequences obtained after direct sequencing of products with those available in the nucleotide databases. The efficiency of each assay for enterovirus identification was evaluated by both distance (Clustal) and similarity (M-NW) indices. Comparative results obtained independently in the three regions showed the highest yield of correlation between nucleotide sequences of all prototype serotypes and the analyzed genotypes in the VP1 region (26/26, 100% Clustal; 22/26, 85% M-NW). Conversely, the VP2 region failed to identify some of the circulating enteroviruses (17/26, 65% Clustal; 16/26, 62% M-NW). Using the RNA polymerase region, sequences from samples and isolates were associated with prototype strains whenever these were available (20/21, 95% Clustal; 12/21, 57% M-NW). These assays were useful for molecular identification of enterovirus directly from samples even when isolation was not possible. Copyright 2001 Wiley-Liss, Inc.
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                Author and article information

                Journal
                ram
                Revista argentina de microbiología
                Rev. argent. microbiol.
                Asociación Argentina de Microbiología (Ciudad Autónoma de Buenos Aires, , Argentina )
                0325-7541
                1851-7617
                March 2008
                : 40
                : 1
                : 41-46
                Affiliations
                [01] orgnameHospital Provincial de Pediatría
                [02] Posadas Misiones orgnameUniversidad Nacional de Misiones orgdiv1Facultad de Ciencias Exactas, Químicas y Naturales orgdiv2Departamento de Microbiología
                [03] Ciudad Autónoma de Buenos Aires orgnameINEI-ANLIS Dr. Carlos G. Malbran orgdiv1Departamento de Virología orgdiv2Servicio de Neurovirosis Argentina
                Article
                S0325-75412008000100009 S0325-7541(08)04000100009
                d08f1179-96c6-48de-9711-46fbc45aca50

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

                History
                : 19 June 2007
                : 18 December 2007
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 44, Pages: 6
                Product

                SciELO Argentina


                Viral meningitis,Brote,ECHO 4,Outbreak,Meningitis viral
                Viral meningitis, Brote, ECHO 4, Outbreak, Meningitis viral

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