Expression of enterovirus 71 capsid protein VP1 in Escherichia coli and its clinical application

The VPl gene of enterovirus 71 (EV71) was synthesized, construct a recombinant plasmid pET15b/VP1 and expressed in E. coli BL21. The recombinant VP1 protein could specifically react with EV71-infected patient sera without the cross-reaction with serum antibodies of coxsackievirus A16 (CA16), A4, A5, B3 and B5 as well as echovirus 6. In acute and convalescent phases, IgM and IgG antibodies of 182 serum samples were detected by ELISA with recombinant VP1 protein as a coated antigen. The results showed that the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of IgM antibodies in serum samples for the diagnosis of EV71 infection were 90.1, 98.4, 98.8 and 88.7%, respectively; similarly, those of IgG antibodies in serum samples were 82.4, 89.1, 91.5 and 78.1%, respectively. Five of 80 samples (6.25%) from CA16infected patients were detected positive by ELISA with recombinant VP1 protein in which indicated the cross reactions and 0 of 5 samples from patients infected with other enteroviruses including CA4, CA5, CB3, CB5 and echovirus 6. Therefore, the recombinant VP1 protein of EV7l may provide a theoretical reference for establishing an effective antibody screening of IgM for EV71-infected patients with clinically suspected hand, foot, and mouth disease (HFMD).

Evaluation of enterovirus 71 immune status in São Paulo state, Brazil / Avaliação da situação imunitária contra enterovírus 71 no estado de São Paulo, Brasil

Antibodies to Enterovirus 71 (EV71) were evaluated in São Paulo State during 1999-2005. The titer of neutralizing antibodies against EV71 was determined by microneutralization assay, and a titer of > 1:8 was defined as indicative of protected immunity. Neutralizing antibodies to EV71 were observed in 12.4 percent (55/442) of sera samples, a low protective rate, suggesting that EV71 infection is uncommon in this region, but that there is a relatively high susceptibility to EV71 related diseases, which is worrying considering the recent Asian outbreaks. Also, a significant location-specific difference in seropositivity was observed. Neutralizing antibodies to EV71 were observed in 8.7 percent (21/241) of São Paulo metropolitan area sera samples, and 16.9 percent (34/201) of the sera samples from other municipalities. A high number of Brazilian residents live in country and coastal areas without adequate access to piped water or sanitation. This situation may contribute to the EV71 dissemination in these zones. The analysis of environmental samples could possibly make a valuable contribution to studies on the epidemiology of EV71.

Anticorpos para Enterovírus 71 (EV71) foram avaliados no Estado de São Paulo durante 1999-2005. O título de anticorpos neutralizantes contra EV71 foi determinado pelo ensaio microneutralização, e um título de > 1:8 foi definido como indicador de imunidade protetora. Anticorpos neutralizantes para EV71 foram observados em 12,4 por cento (55/442) das amostras de soro, uma baixa taxa de proteção, sugerindo que a infecção pelo EV71 é incomum nesta região e que existe alta susceptibilidade a doenças relacionadas ao EV71, o que é preocupante considerando os recentes surtos asiáticos. Ainda, foi observada diferença significativa na soropositividade em relação à localização, onde 8,7 por cento (21/241) e 16,9 por cento (34/201) das amostras provenientes da região metropolitana de São Paulo, e demais municípios, respectivamente, apresentaram anticorpos neutralizantes para EV71. Um grande número de brasileiros vive em áreas rurais e à beira-mar, sem acesso adequado à água encanada ou saneamento. Essa situação pode contribuir para a disseminação de EV71 nessas regiões. A análise de amostras ambientais poderia gerar contribuição valiosa para estudos sobre a epidemiologia da EV71.