Padronização de teste para quantificar anticorpos anti-poliovírus utilizando vírus vacinal / Standardization of a test for quantifying the antibodies against poliovirus using the vaccine virus

A poliomielite é uma doença endêmica no Afeganistão e no Paquistão, apesar dos esforços para ser erradicada, representando uma ameaça para outros países principalmente devido às viagens internacionais. A Organização Mundial da Saúde (OMS) tem como objetivo erradicar a poliomielite causada pelo poliovírus selvagem no mundo. O requisito essencial para a erradicação da poliomielite é a eliminação da cepa do poliovírus selvagem, que é empregada no teste padrão-ouro. Com o intuito de auxiliar na erradicação do poliovírus selvagem, o objetivo deste estudo foi modificar o teste padrão-ouro usando o poliovírus derivado da vacina oral atenuada. Foram testados 63 soros pelo ensaio de neutralização utilizando-se antígenos vacinais. A concordância do sorotipo 1 (k=0,74) foi considerada substancial, enquanto o sorotipo 2 (k=1,00) e sorotipo 3 (k= 0,95) foram consideradas quase perfeitas. A sensibilidade dos testes de soroneutralização utilizando os sorotipos 1, 2 e 3 foi de 94,83%, 100,00% e 100,00%, respectivamente. Em conclusão, o ensaio com antígenos vacinais pode ser usado como procedimento laboratorial seguro, especialmente em estudos de vigilância em larga escala.

Poliomyelitis is an endemic disease in Afghanistan and Pakistan in despite of the efforts to eradicate it, and it represents a threat to other countries mainly due to the international trips. The World Health Organization (WHO) aims at eradicating the polio disease worldwide. An essential requirement for eradicating the poliomyelitis is the elimination of the wild poliovirus strain, which is employed in the gold standard test. As a support for the eradication of wild poliovirus, the present study aimed at modifying the gold standard test by using poliovirus derived from the oral attenuated vaccine. Sixty-three sera samples were tested by neutralization assay using vaccine antigens. The degree of agreement of the serotype 1 (k=0.74) was considered substantial, while the serotype 2 (k=1.00) and 3 (k= 0.95) showed almost perfect agreement. The sensitivity of serotypes 1, 2 and 3 was 94.83%, 100.00% and 100.00%, respectively. In conclusion, the assay with the vaccine antigens can be used as a safe application, especially for large-scale surveillance studies.

Comparative study of two extraction methods for enteric virus recovery from sewage sludge by molecular methods

The aim of this study was to compare two nucleic acid extraction methods for the recovery of enteric viruses from activated sludge. Test samples were inoculated with human adenovirus (AdV), hepatitis A virus (HAV), poliovirus (PV) and rotavirus (RV) and were then processed by an adsorption-elution-precipitation method. Two extraction methods were used: an organic solvent-based method and a silica method. The organic-based method was able to recoup 20 percent of the AdV, 90 percent of the RV and 100 percent of both the PV and HAV from seeded samples. The silica method was able to recoup 1.8 percent of the AdV and 90 percent of the RV. These results indicate that the organic-based method is more suitable for detecting viruses in sewage sludge.

Guillain Barre syngdrome: the leading cause of acute flaccid paralysis in hazara division

Acute flaccid paralysis [AFP] can be caused by a number of conditions. A common preventable cause is poliomyelitis which is still being reported in Pakistan, Guillain Barre syndrome [GBS], also know as Acute Inflammatory Demyelinating Polyneuropathy, is another common cause of acute flaccid paralysis. It is important to recognize GBS in childhood as parents consider all acute flaccid paralysis to be due to poliomyelitis. The present study was designed to know the frequency of different causes of acute flaccid paralysis in Hazara division. Hazara division during the period January 2003 to December 2004. acute flaccid paralysis was flaccid paralysis was investigated by appropriate laboratory tests, such as serum electrolytes, virus and other enteroviruses. Diagnosis of Poliomyelitis was confirmed by stool testing for poliovirus. 74 patients presented with AFP during the study period. 36 were male and 38 were female. Guillain Barre syndrome and enteroviral encephalopathy were the two leading causes of acute flaccid paralysis. Majority of the cases were reported from Mansehra district. Children of age groups 12 to 24 months and > 96 months constituted the majority [20% each]. Guillain Barre syndrome was the leading cause of acute flaccid paralysis reported from various parts of Hazara division

Investigation of circulating anti-coxsackie B, anti- polio and anti-adeno IgG in newly diagnosed T1DM children

Viruses may be involved in the pathogenesis of Type 1 Diabetes Mellitus [T1DM], either through direct beta-cell infection or as triggers of autoimmunity. To investigate the presence of specific anti- viral IgG antibodies for Coxsackie virus type B [CVB5], Poliovirus, and Adenovirus which proposed to be involved in the etiology of T1DM. A total of 60 Iraqi T1DM children were included in the present study. They were new onset of the disease [diagnosis was from one week up to five months]. For the purpose of comparisons, 50 apparently healthy control subjects were selected. Serum IgG against Coxsackie virus type B[5], Adenovirus type 3, 4, and 7, and Poliovaccin Trivalent were detected quantitatively with an indirect ELISA. High proportion of anti-CVB5 IgG [20%][p<0.05] and anti- Polio IgG [31.67%] were found in T1DM children compared to controls [8%, 26% respectively], while anti- Adeno IgG were detected in diabetic patients only [6.67%][p<1.0001]

Detección de Poliovirus en Aguas Residuales de Armenia, Colombia / Detecting poliovirus in wastewater from Armenia, Colombia

OBJETIVOS: Implementar un método para la vigilancia ambiental de poliovirus circulante a partir de aguas residuales. MÉTODOS: Se colectaron 6 muestras de aguas residuales en los sitios finales de descarga del acueducto de Armenia, Quindío. Los virus fueron concentrados y extraídos por un método que usa el polietilenglicol y cloruro de sodio como agentes concentradores. La identificación de poliovirus se realizó mediante la técnica de reacción en cadena de la polimerasa empleando cebadores específicos de grupo, de serotipo y de cepa vacunal sabin. RESULTADOS: Se demostró la eficiencia de los métodos de detección viral empleados. Se encontró la presencia de poliovirus en 5 de las 6 (83 %) muestras colectadas. La identificación serotipo-específica reveló la presencia de poliovirus tipo 1 y 3 en las muestras estudiadas. Todos los poliovirus detectados resultaron ser del tipo vacunal lo cual aporta una evidencia más a favor de la no circulación de poliovirus salvaje en la región. CONCLUSIÓN El sistema de vigilancia a partir de aguas residuales puede ser una herramienta sensible para la detección de la circulación de poliovirus.

Detecting polio through surveillance for acute flaccid paralysis (AFP).

Accurate surveillance for polio is essential for eradication. Surveillance systems for polio has been developed under the guidance of the global polio eradication initiative. Surveillance of cases of acute flaccid paralysis among children less than 15 years of age is a key component for a well functioning polio surveillance system. The surveillance system works through a network of surveillance medical officers, the responsibility of them lies in assisting the health services departments of all states and maintaining a network of acute flaccid paralysis reporting sites and rapidly investigating the cases. Surveillance activities begin when a child comes in contact with a healthcare provider who in turn informs the officer in charge of acute flaccid paralysis surveillance. The goal of the polio network laboratories is to provide accurate and timely results of wild poliovirus detection in stool samples of cases of acute flaccid paralysis. Strong linkages have been established between the acute flaccid paralysis surveillance system and the laboratory network. Laboratories complete poliovirus isolation and if poliovirus is isolated, these are submitted for intratypic differentiations. Acute flaccid paralysis surveillance in India has demonstrated that the eradication activities implemented in India led to dramatic reduction and restriction in the number of cases and geographic spread of poliovirus transmission.

Laboratory acute flaccid paralysis surveillance in Malaysia: a decade of commitment to the WHO global polio eradication initiative.

The Institute for Medical Research, Malaysia, was designated the National Reference Laboratory for Poliomyelitis Eradication (NRLPE) in 1992. Since then, our Polio Laboratory has collaborated actively with the Disease Control Division, Ministry of Health (MOH), Malaysia and WHO towards achieving polio eradication. Since 1992, the NRLPE has investigated 1,063 stool specimens from 641 acute flaccidparalysis (AFP) cases. One hundred and one enteroviruses were isolated from these specimens. Positive cell cultures were confirmed by microneutralization assay using standard WHO antisera. All enterovirus isolates were sent to the Victorian Infectious Disease Reference Laboratory in Melbourne, Australia, for further identification and poliovirus intratypic differentiation. Thirty-one out of these 101 virus isolates (30%) were polioviruses (PV) and the remaining 70 (70%) were non-polio enteroviruses (NPEV) which included coxsackie B viruses, echoviruses and enterovirus 71. Three of the poliovirus isolates were wild-type polioviruses isolated in 1992 which were the last wild-type polioviruses isolated in Malaysia. The rest were vaccine-related Sabin-like strains. Monthly reports of the virological investigation of AFP cases are sent to WHO and to the MOH, AFP control committee. The NRLPE continues to play an integral role in AFP surveillance and is committed to the WHO's goal of global polio eradication by the year 2005.

Detection of poliovirus, hepatitis A virus and rotavirus from sewage and water samples.

A modified adsorption-elution technique for concentration of enteric viruses from sewage and water samples was developed. The viruses in water were concentrated by negatively charged membrane filtration, eluted with 2.9% tryptose phosphate broth containing 6% glycine pH 9.0, and reconcentrated using centrifugation by a speedVac concentrator. The presence of poliovirus, hepatitis A virus (HAV) RNA, and rotavirus antigen was determined by cell culture isolation, nested polymerase chain reaction (nested PCR), and enzyme-linked immunosorbent assay (ELISA), respectively. A total of 100 sewage and water samples were collected from various sources in congested communities in Bangkok, concentrated and examined for those enteric viruses. Of 20 surface water samples from canals which located near sewage drains, 15% were positive for HAV RNA by nested PCR. Of 48 domestic sewage samples from man-holes of underground sewers, 8% were positive for rotavirus antigen by ELISA. Even though the samples were concentrated 256-2,000 fold, poliovirus was not found by isolation in cell culture.

Ausencia de detección de enterovirus en bivalvos Anadara tuberculosa (Bivalvia: Arcidae) por contaminación química en el Pacífico de Costa Rica / Absence of detection of enterovirus in bivalvos Anadara tuberculosa (Bivalvia: Arcidae) by chemical contamination in the Pacific of Costa Rica

Anadara tuberculosa es uno de los moluscos más abundantes de importancia comercial en Costa Rica. Su hábitat acuático es una fuente potencial de contaminación fecal y química para el ser humano. El objetivo de este trabajo fue determinar enterovirus, especialmente poliovirus y el virus de la hepatitis A y contaminación química como nitratos y sulfatos en las carnes y los líquidos internos...

Caracterización intratípica de Poliovirus por la técnica de reacción en cadena de la polimerasa

Se introdujo la técnica de la reacción en cadena de la polimerasa para la caracterización intratípica de Poliovirus. Se usaron cebadores que sólo promueven la ampliación de las cepas vacunales de Sabin, comprobada por corrida electroforética de los productos de ADN amplificados (Sabin 1-97 pb, Sabin 2-71 pb, Sabin 3-44 pb) y cuya especificidad se verificó satisfactoriamente. Se estudiaron por esta técnica 23 cepas cubanas de Poliovirus aisladas e identificadas en el Laboratorio de Enterovirus del Instituto de Medicina Tropical "Pedro Kourí" de 1993 a 1994, y todas resultaron ser del tipo vacunal. Se observó cómo el Poliovirus vacunal de Sabin puede ser causa de meningoencefalitis viral como complicación neurológica más leve. Este estudio aportó una evidencia más a favor de la no circulación del Poliovirus salvaje en Cuba.

The polimerase chain reaction techniques was introduced for the intratypic characterization of Poliovirus. Primers were used only to promote the amplification of the Sabin vaccine strains proved by electrophoretic run of the amplified DNA products (Sabin 1 - 97 pb, Sabin 2 - 71 pb, Sabin 3 - 44 pb) and whose specificity was satisfactorily verified. 23 Cuban poliovirus strains isolated and identified at the Laboratory of Enterovirus of the "Pedro Kourí" Tropical Medicine Institute from 1993 to 1994 were studied by this technique. All of them were of the vaccine type. It was observed how the Sabin vaccine poliovirus may be the cause of viral meningoencephalitis as a milder neurological complication. Tghis study provided one more evidence about the non circulation of the wild poliovirus in Cuba.

Caracterización molecular de enterovirus no-polio procedentes de casos con parálisis residual / Molecular characterization of non-polio enteroviruses from residual paralysis cases. Colombia, 1992 - 1995

Las parálisis fláccidas agudas (PFA) tienen una amplia variedad de orígenes y de agentes causales: físicos (traumas), fisiopatológicos (Accidente cerebro vascular (ACV), tóxicos (drogas o químicos) e infecciosos (bacterias y virus). Entre estos últimos, el virus salvaje de la poliomielitis y el enterovirus 71 (EV71), son los agentes virales más frecuentes. Con la no-detección de poliovirus salvaje autóctono como agente causal de enfermedad paralítica en Colombia desde junio de 1991 y aislamientos de virus No-polio en el 20.84 por ciento del total de casos de PFA notificados anualmente, se quiso conocer el papel que jugan los enterovirus en la incidencia de parálisis fláccida Aguda y la dinámica de circulación y distribución de los mismos en Colombia, para lo cual, se revisó la base de datos epidemiológicos y clínicos de los casos notificados al programa de erradicación de la poliomielitis en Colombia a partir del 1º de enero de 1992 al 31 de diciembre de 1995. Se clasificaron los casos con base en la presencia de parálisis residual y la entidad clínica de descarte según valoración y clasificación realizada por el Grupo de control de patologías del Ministerio de Salud, el Programa Ampliado de Inmunizaciones (PAI), la Organización Panamericana de la Salud y la Organización Mundial de la Salud (OPS/OMS). Durante estos cuatro años, el Sistema de Vigilancia Epidemiológica de las parálisis fláccideas, notificó 856 casos en menores de 15 años, de los cuales, 346 (40.42 por ciento) presentaron paralisis residual, 331 (95.6 por ciento) tuvieron muestras de heces para estudio virológico. Se seleccionaron los casos con estudio virológico para Enterovirus (incluyendo poliovirus) y se encontró que las patologías más frecuentes fueron (Síndrome de Guillaín Barré, neuropatía periférica, encefalitis y meningitis virales, hemiplejía infantil aguda, esclerosis múltiple, mielitis transversa y mielopatías, miositis, polimiositis, monoparesia y dermatomiositis. De estos, 69 casos (20.84 por ciento) tuvieron aislamiento de virus No polio. En 16 casos (4.8 por ciento el aislamiento fue un poliovirus vacunal, 5 de los cuales (1.2 por ciento) se asociaron a poliomielitis paralítica post-vacunal. Se realizó identificación de serotipos mediante neutralización con mezclas de antisueros anti-enterovirus de Lim & Benyesh-Melnick -LBM- (40,41) y caracterización molecular mediante reacción en cadena de la polimerasa -PCR- utilizando primers complementarios a la región VP1 del...

Virologic surveillance of poliomyelitis in Delhi.

OBJECTIVE: Virologic surveillance of poliomyelitis to monitor the transmission of wild polio virus in the community. STUDY AREA: All major hospitals of Delhi and surrounding area. METHODS: Stool samples were collected from 1221 cases of acute flaccid paralysis during 1992-1994 and were subjected to virus isolation on RD and HEp2 cell line. Viruses isolated were analyzed further by microneutralization test using polio and nonpolio antisera. The polio isolates were further characterized as vaccine or wild type using ELISA and probe technology. RESULTS: Out of the 1221 cases tested, virus was isolated in 57.4%. Among the virus positive cases, polio was isolated in 57% and in 43% non polio entero viruses were detected. The most prevalent was polio virus type 1. Most of the strains were wild type. CONCLUSION: Wild polio virus was prevailing in the community under study between the years 1992-1994.

Absence of wild poliovirus circulation among healthy children in a rural area with high oral poliovirus vaccination coverage.

Poliovirus circulation in a rural community was studied by a stool sample survey. No acute paralytic poliomyelitis case had been reported from the study area during the previous 5 months. Immunization coverage in age groups 7 to 12 months and 12 to 60 months was 95.8 and 94 per cent, respectively. Of the 257 children from whom stool samples were collected (about 6% of the child population), 161 (62.6%) were positive for virus isolation. Poliovirus was isolated from 60 (23.3%) children. All three poliovirus types were detected (41 type 1, 16 type 2 and 3 type 3). Intratypic differentiation tests classified these isolates as vaccine-like. Among the children excreting poliovirus, the proportion of those who did not receive polio vaccine within 30 days prior to the sample collection was 46.3, and 68.7 per cent for poliovirus type 1 and 2, respectively. It was concluded that these poliovirus excreting children were infected by the vaccine strains circulating in the environment. The survey showed that wild poliovirus was not detectable within five months after the last case of acute poliomyelitis. Displacement of the wild virus from the environment and circulation of vaccine virus was achieved by high vaccination coverage in this area.

Human enterovirus infection in stray dogs: some aspects of interest to public health

Foram estudados 212 caes errantes, capturados em areas perifericas no municipio de Sao Paulo, com o objetivo de investigar o possivel papel desses animais como reservatorios de enterovirus humanos. Os animais capturados foram divididos em 19 grupos compostos por 10 a 20 animais cada um. Foram efetuadas tentativas de isolamento em amostras de fezes em 126/212 caes, obtendo-se resultados positivos em 12 deles, assim distribuidos: poliovirus tipo 1 (dois caes), poliovirus tipo 3 (um cao), echovirus tipo 7 (oito caes) e echovirus tipo 15 (um cao). Dos 12 animais infectados, quatro apresentaram anticorpos neutralizantes homotipicos com titulos>-16. Todos os 212 caes foram submetidos a pesquisa de anticorpos neutralizantes contra enterovirus humanos...

Temporal association between the isolation of Sabin-related poliovirus vaccine strains and the Guillain-Barre syndrome

Thirty eigth paralysis classified as Guillain-Barre syndrome (GBS) in Brazil were analysed. In all these cases Sabin-related poliovirus vaccine strains were isolated. In most of the cases the last vaccine dose was given months or years before the onset of GBS, suggesting a persistent infection or transmission of the Sabin-related strains to the patients...