Genomic Characterization of the Historical Smallpox Vaccine Strain Wyeth Isolated from a 1971 Seed Vial.

The Wyeth strain of vaccinia virus (VACV) produced by Wyeth Pharmaceuticals was supposedly used to manufacture the old freeze-dried American smallpox vaccine, Dryvax, until its discontinuation in 2008. Although the genomic sequences of numerous Dryvax clones have been reported, data on VACV-Wyeth genomes are still lacking. Genomic analysis of old VACV strains is relevant to understand the evolutionary relationships of smallpox vaccines, particularly with the recent resumption of smallpox vaccination in certain population groups as an attempt to control the worldwide monkeypox outbreak. Here we analyzed the complete genome sequences of three VACV-Wyeth clonal isolates obtained from a single seed vial donated to the Brazilian eradication program in the 1970s. Wyeth clones show >99.3% similarity to each other and >95.3% similarity with Dryvax clones, mapping together in clade I of the vaccinia group. Although the patterns of SNPs and INDELs comparing Dryvax and Wyeth clones are overall uniform, important differences were detected particularly at the ends of the genome. In addition, we detected recombinant events of clone Wyeth A111 and the Dryvax clone Acam2000, suggesting that other regions of the genomes may have similar patchy patterns of recombination. A small-scale serological survey using VACV-Wyeth as antigen in ELISA assays revealed that 63 of the 65 individuals born before the end of smallpox vaccination in Brazil still have anti-VACV IgG antibodies, demonstrating the usefulness of the VACV-Wyeth strain in future extended serological studies of the Brazilian population.

Serodiagnosis of neurocysticercosis in patients with epileptic seizure using ELISA and immunoblot assay

Sera from 88 patients from Santa Catarina and São Paulo states of Brazil, with epileptic seizures who underwent cerebral computed tomography (CT) were analyzed for the detection of antibodies to T. solium cysticercus by ELISA and Immunoblot (IB) with the following antigens: Taenia solium cysticercus total saline (Tso), Taenia crassiceps cysticercus vesicular fluid (Tcra-vf) and T. crassiceps cysticercus glycoproteins (Tcra-gp). ELISA carried out with Tso, Tcra-vf and Tcra-gp antigens showed 95 percent, 90 percent and 80 percent sensitivities, respectively, and 68 percent, 85 percent and 93 percent specificities, respectively. In the epileptic patients group, ELISA positivity was 30 percent, 51 percent and 35 percent with Tso, Tcra-vf and Tcra-gp antigens respectively. Considering the IB as the confirmatory test, the positivity was 16 percent (14/88) in the epileptic patients total group and 22 percent (12/54) in the epileptic patients with positive CT and signals of cysticercosis. We found a significant statistical correlation among ELISA or IB results and the phase of the disease when any antigens were used (p < 0.05). We emphasize the need to introduce in the laboratory routine the search for neurocysticercosis (NC) in patients presenting with epileptic seizures because of the high risk of acquiring NC in our region and its potential cause of epilepsy.

Amostras de soro de 88 pacientes dos Estados de Santa Catarina e São Paulo, Brasil, com crises epilépticas e que se submeteram a exame de Tomografia Computadorizada (TC), foram examinadas para detecção de anticorpos anti-cisticercos de Taenia solium por meio de ELISA e Immunoblot (IB) utilizando-se os seguintes antígenos: extrato salino total de cisticercos de T. solium (Tso); líquido vesicular de Taenia crassiceps (Tcra-vf) e glicoproteínas purificadas de cisticercos de T. crassiceps (Tcra-gp). Os resultados de ELISA com os antígenos Tso, Tcra-vf e Tcra-gp mostraram 95 por cento, 90 por cento e 80 por cento de sensibilidade, respectivamente, e 68 por cento, 85 por cento e 93 por cento de especificidade, respectivamente. No grupo de pacientes epilépticos, a positividade do ELISA foi 30 por cento, 51 por cento e 35 por cento com os antígenos Tso, Tcra-vf e Tcra-gp, respectivamente. Considerando o IB como teste confirmatório, a positividade foi de 16 por cento (14/88) no grupo total de pacientes epilépticos e 22 por cento (12/54) no grupo de pacientes epilépticos com TC positiva e sinais clínicos compatíveis com neurocisticercose. Foi encontrada correlação estatística significativa entre os resultados de ELISA ou IB e a fase da doença com quaisquer dos antígenos utilizados (p < 0,05). Os resultados indicam a necessidade de introduzir na rotina dos laboratórios o diagnóstico de neurocisticercose nos pacientes com convulsões epilépticas devido ao elevado risco de aquisição da cisticercose em nossa região e sua participação na etiologia da epilepsia.

Serodiagnosis of neurocysticercosis in patients with epileptic seizure using ELISA and immunoblot assay.

Sera from 88 patients from Santa Catarina and São Paulo states of Brazil, with epileptic seizures who underwent cerebral computed tomography (CT) were analyzed for the detection of antibodies to T. solium cysticercus by ELISA and Immunoblot (IB) with the following antigens: Taenia solium cysticercus total saline (Tso), Taenia crassiceps cysticercus vesicular fluid (Tcra-vf) and T. crassiceps cysticercus glycoproteins (Tcra-gp). ELISA carried out with Tso, Tcra-vf and Tcra-gp antigens showed 95%, 90% and 80% sensitivities, respectively, and 68%, 85% and 93% specificities, respectively. In the epileptic patients group, ELISA positivity was 30%, 51% and 35% with Tso, Tcra-vf and Tcra-gp antigens respectively. Considering the IB as the confirmatory test, the positivity was 16% (14/88) in the epileptic patients total group and 22% (12/54) in the epileptic patients with positive CT and signals of cysticercosis. We found a significant statistical correlation among ELISA or IB results and the phase of the disease when any antigens were used (p < 0.05). We emphasize the need to introduce in the laboratory routine the search for neurocysticercosis (NC) in patients presenting with epileptic seizures because of the high risk of acquiring NC in our region and its potential cause of epilepsy.