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      Respuesta de anticuerpos IgG contra fracciones de Giardia duodenalis en individuos infectados Translated title: IgG antibody response against fractions of Giardia duodenalis in infected individuals

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          Abstract

          La Giardiasis es una infección causada por Giardia duodenalis, transmitida por el agua o alimentos contaminados con quistes del parásito, principalmente en escolares que habitan zonas rurales. Se conoce poco sobre la respuesta de anticuerpos específicos, la identificación de antígenos permitirá el diseño de pruebas para evaluar seroprevalencia. En este estudio fueron identificadas proteínas para la respuesta IgG anti-Giardia por inmunodetección. Fueron evaluados ochenta individuos entre 4-80 años, de ambos sexos, de comunidades rurales, fueron clasificados según la presencia o ausencia de quistes del parásito en las heces. Las proteínas fueron obtenidas a partir de un extracto soluble y fracciones por centrifugación diferencial de la cepa WB, analizadas por electroforesis en geles de poliacrilamida, transferidas a membranas de nitrocelulosa y probadas por Western Blot para la respuesta IgG anti-Giardia. Fue observada una prevalencia general entre 9 y 12% de parasitosis intestinales, con 22% de individuos positivos para Giardia duodenalis, 46% negativos en heces y 32% positivos para Giardia duodenalis y otras parasitosis. El perfil de proteínas reveló bandas entre 190-15kDa, donde fueron reconocidas para la respuesta específica de anticuerpos IgG, proteínas en la fracción 2 de 190, 176 y 51kDa, en la fracción 3 una proteína de 23kDa. La identificación de proteínas reactivas para la respuesta IgG en las fracciones 2 y 3 permitirá el diseño de pruebas serológicas más específica para discriminar seropositivos de seronegativos.

          Translated abstract

          Giardiasis is an infection caused by Giardia duodenalis, transmitted by consuming water or food contaminated with Giardia cysts, mainly in children living in rural areas. Little is known about proteins associated to the specific antibody response, the identification of specific antigens is necessary for studies of seroprevalence. The aim of this study was to select antigenic proteins from soluble extract or fractions of the parasite for the design of specific diagnostic tests. Were evaluated eighty individuals between 4-80 years, from rural communities were classified according to the presence or absence of cysts in the stool. The soluble extract and the fractions were obtained from WB (ATCC) strain of Giardia sp by differential centrifugation, analyzed by polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes and finally tested by Western Blots for the IgG antibody response. The general prevalence was between 9-12%, and 22% of them were positive for G. duodenalis, 46% of individuals were negative in feces; and 32% were positive for G. duodenalis and other parasites. The protein profile revealed several bands from 190-15kDa; in which were recognized mainly proteins of 190, 176 and 51kDa in the fraction 2, and a protein of 23kDa in the fraction 3. The infection by G. duodenalis represents a human public health, and the identification of reactive proteins for the IgG response in the fractions 2 and 3, would allow us the design of serological test more specific, in order to discriminate the seropositive individuals from those seronegative.

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          Axenic culture of Giardia lamblia in TYI-S-33 medium supplemented with bile

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            Comparative genomic analyses of freshly isolated Giardia intestinalis assemblage A isolates

            Background The diarrhea-causing protozoan Giardia intestinalis makes up a species complex of eight different assemblages (A-H), where assemblage A and B infect humans. Comparative whole-genome analyses of three of these assemblages have shown that there is significant divergence at the inter-assemblage level, however little is currently known regarding variation at the intra-assemblage level. We have performed whole genome sequencing of two sub-assemblage AII isolates, recently axenized from symptomatic human patients, to study the biological and genetic diversity within assemblage A isolates. Results Several biological differences between the new and earlier characterized assemblage A isolates were identified, including a difference in growth medium preference. The two AII isolates were of different sub-assemblage types (AII-1 [AS175] and AII-2 [AS98]) and showed size differences in the smallest chromosomes. The amount of genetic diversity was characterized in relation to the genome of the Giardia reference isolate WB, an assemblage AI isolate. Our analyses indicate that the divergence between AI and AII is approximately 1 %, represented by ~100,000 single nucleotide polymorphisms (SNP) distributed over the chromosomes with enrichment in variable genomic regions containing surface antigens. The level of allelic sequence heterozygosity (ASH) in the two AII isolates was found to be 0.25–0.35 %, which is 25–30 fold higher than in the WB isolate and 10 fold higher than the assemblage AII isolate DH (0.037 %). 35 protein-encoding genes, not found in the WB genome, were identified in the two AII genomes. The large gene families of variant-specific surface proteins (VSPs) and high cysteine membrane proteins (HCMPs) showed isolate-specific divergences of the gene repertoires. Certain genes, often in small gene families with 2 to 8 members, localize to the variable regions of the genomes and show high sequence diversity between the assemblage A isolates. One of the families, Bactericidal/Permeability Increasing-like protein (BPIL), with eight members was characterized further and the proteins were shown to localize to the ER in trophozoites. Conclusions Giardia genomes are modular with highly conserved core regions mixed up by variable regions containing high levels of ASH, SNPs and variable surface antigens. There are significant genomic variations in assemblage A isolates, in terms of chromosome size, gene content, surface protein repertoire and gene polymorphisms and these differences mainly localize to the variable regions of the genomes. The large genetic differences within one assemblage of G. intestinalis strengthen the argument that the assemblages represent different Giardia species. Electronic supplementary material The online version of this article (doi:10.1186/s12864-015-1893-6) contains supplementary material, which is available to authorized users.
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              Aprendizaje de la parasitología basado en problemas

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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                s
                Salus
                Salus
                Universidad de Carabobo
                1316-7138
                August 2016
                : 20
                : 2
                : 13-17
                Affiliations
                [1 ] Instituto de Inmunología Dr. Nicolás E. Bianco C Venezuela
                [2 ] Universidad Central de Venezuela Venezuela
                Article
                S1316-71382016000200004
                978b27cf-189d-4a48-b3f8-a26d0eb6cf82

                http://creativecommons.org/licenses/by/4.0/

                History
                Product

                SciELO Venezuela

                Self URI (journal page): http://www.scielo.org.ve/scielo.php?script=sci_serial&pid=1316-7138&lng=en
                Categories
                HEALTH CARE SCIENCES & SERVICES
                HEALTH POLICY & SERVICES

                Health & Social care,Public health
                Giardiasis,Western Blot en giardasis,anticuerpos IgG para G. duodenalis,Giardiasis` Western Blot,G. duodenalis´ IgG-antibodies

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