Objetivos. Diseñar y evaluar una proteína multiepítope como candidato a vacuna contra la enfermedad de Carrión. Materiales y métodos. Mediante herramientas bioinformáticas se seleccionó epítopes de proteínas de membrana externa y se diseñó una proteína multiepítope. El gen de la proteína multiepítope fue subclonado en el plásmido de expresión pET28b y transformado en E. coli BL21 pLys. La proteína multiepítope fue expresada usando isopropil-β-D-1-tiogalactopiranósido y purificada usando resina. Esta proteína purificada fue utilizada para inmunizar ratones BALB/c y se obtuvo anticuerpos policlonales. Se realizaron ensayos de invasión in vitro usando una cepa de Bartonella bacilliformis (B. bacilliformis) a eritrocitos humanos. Resultados. La proteína multiepítope M1 presenta epítopes conservados entre aislamientos de B. bacilliformis, no tóxicos, no homólogos a proteínas humanas y superficiales. Los ratones inmunizados presentaron niveles de anticuerpos IgG capaces de reducir in vitro la tasa de invasión de B. bacilliformis a eritrocitos humanos. Conclusiones. La proteína multiepítope M1 podría servir como candidato a vacuna contra la enfermedad de Carrión; sin embargo, se requiere de más estudios para caracterizar el uso de este antígeno como vacuna.
Objectives: to identify the genotypes of Hepatitis Delta Virus (HDV) circulating in populations of the inter-Andean valley of Huanta and three indigenous peoples of the Peruvian Amazon. Materials and Methods: Observational and cross-sectional study, from 582 reactive samples for anti-HBc-HBV antibodies in inhabitants of the province of Huanta and the Amazonian towns of Matses, Kandozi and Chapra (Loreto). Analysis was performed for HDV infection markers: anti-HDV IgM and anti-HDV IgG by ELISA using Wantai brand kits. Anti-HDV ELISA positive samples were processed with the nRT-PCR method for the detection of HDV RNA. HDV genotype was determined by direct Sanger-type sequencing and phylogenetic analysis of the R0 fragment. 111 reference sequences from GenBank were used. The 42 sequences of the study were edited, assembled and cut with the programs Chromas 2.6.5, Bioedit v7.2, ClustalW v.1.6 of Mega v.7.0 and the Gblocks server. Phylogenetic and evolutionary analysis was performed with the following software: Beast V2.5.2, Jmodeltest v2.1.10, Tracer v1.7.1, Tree Annotator and Figtree v1.4.4. The Bayesian Yule and Birth Death skyline serial models were used, the MCMC at 30 and 80 million respectively, with the relaxed uncorrelated Exponential molecular clock. Summary and central tendency measures were calculated using the program in STATA 14.0. Results.- The mean age was 38 years (0 to 86 years), 52.75% (N=307) were women. 582 blood samples positive for anti-HBc were analyzed for anti-HVD antibodies using the ELISA method, with 101 positive samples being found. HDV RNA was detected in 49.50% of the anti-HDV ELISA reactive samples. Phylogenetic analysis determined the presence of genotype 3. Conclusions: The presence of HDV genotype 3 in Andean and Amazonian communities of Peru is evidenced. Keywords: Hepatitis D; Indigenous population; Indigenous communities (source: DeCS BIREME).
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