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  1. Article: A Chikungunya Virus Multiepitope Recombinant Protein Expressed from the Binary System Insect Cell/Recombinant Baculovirus Is Useful for Laboratorial Diagnosis of Chikungunya

    Silva, Leonardo Assis da / Lima, Monique da Rocha Queiroz / de Camargo, Brenda Rabello / Guimarães, Dyeferson Kened da Silva Coelho / Barbastefano, Anabele Azevedo Lima / Lima, Raquel Curtinhas de / Damasco, Paulo Vieira / Cunha, Rivaldo Venâncio da / de Souza, Luiz José / de Azeredo, Elzinandes Leal / de-Oliveira-Pinto, Luzia Maria / Nagata, Tatsuya / Ardisson-Araújo, Daniel M. P. / dos Santos, Flavia Barreto / Morais Ribeiro, Bergmann

    Microorganisms. 2022 July 18, v. 10, no. 7

    2022  

    Abstract: Chikungunya virus (CHIKV) is an arbovirus currently distributed worldwide, causing a disease that shares clinical signs and symptoms with other illnesses, such as dengue and Zika and leading to a challenging clinical differential diagnosis. In Brazil, ... ...

    Abstract Chikungunya virus (CHIKV) is an arbovirus currently distributed worldwide, causing a disease that shares clinical signs and symptoms with other illnesses, such as dengue and Zika and leading to a challenging clinical differential diagnosis. In Brazil, CHIKV emerged in 2014 with the simultaneous introduction of both Asian and East/Central/South African (ECSA) genotypes. Laboratorial diagnosis of CHIKV is mainly performed by molecular and serological assays, with the latter more widely used. Although many commercial kits are available, their costs are still high for many underdeveloped and developing countries where the virus circulates. Here we described the development and evaluation of a multi-epitope recombinant protein-based IgG-ELISA (MULTREC IgG-ELISA) test for the specific detection of anti-CHIKV antibodies in clinical samples, as an alternative approach for laboratorial diagnosis. The MULTREC IgG-ELISA showed 86.36% of sensitivity and 100% of specificity, and no cross-reactivity with other exanthematic diseases was observed. The recombinant protein was expressed from the binary system insect cell/baculovirus using the crystal-forming baculoviral protein polyhedrin as a carrier of the target recombinant protein to facilitate recovery. The crystals were at least 10 times smaller in size and had an amorphous shape when compared to the polyhedrin wild-type crystal. The assay uses a multi-epitope antigen, representing two replicates of 18 amino acid sequences from the E2 region and a sequence of 17 amino acids from the nsP3 region of CHIKV. The recombinant protein was highly expressed, easy to purify and has demonstrated its usefulness in confirming chikungunya exposure, indeed showing a good potential tool for epidemiological surveillance.
    Keywords Baculoviridae ; Chikungunya virus ; antigens ; arboviruses ; cross reaction ; dengue ; insects ; monitoring ; recombinant proteins ; Brazil
    Language English
    Dates of publication 2022-0718
    Publishing place Multidisciplinary Digital Publishing Institute
    Document type Article
    ZDB-ID 2720891-6
    ISSN 2076-2607
    ISSN 2076-2607
    DOI 10.3390/microorganisms10071451
    Database NAL-Catalogue (AGRICOLA)

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  2. Article: A Chikungunya Virus Multiepitope Recombinant Protein Expressed from the Binary System Insect Cell/Recombinant Baculovirus Is Useful for Laboratorial Diagnosis of Chikungunya.

    Silva, Leonardo Assis da / Lima, Monique da Rocha Queiroz / de Camargo, Brenda Rabello / Guimarães, Dyeferson Kened da Silva Coelho / Barbastefano, Anabele Azevedo Lima / Lima, Raquel Curtinhas de / Damasco, Paulo Vieira / Cunha, Rivaldo Venâncio da / de Souza, Luiz José / de Azeredo, Elzinandes Leal / de-Oliveira-Pinto, Luzia Maria / Nagata, Tatsuya / Ardisson-Araújo, Daniel M P / Dos Santos, Flavia Barreto / Morais Ribeiro, Bergmann

    Microorganisms

    2022  Volume 10, Issue 7

    Abstract: Chikungunya virus (CHIKV) is an arbovirus currently distributed worldwide, causing a disease that shares clinical signs and symptoms with other illnesses, such as dengue and Zika and leading to a challenging clinical differential diagnosis. In Brazil, ... ...

    Abstract Chikungunya virus (CHIKV) is an arbovirus currently distributed worldwide, causing a disease that shares clinical signs and symptoms with other illnesses, such as dengue and Zika and leading to a challenging clinical differential diagnosis. In Brazil, CHIKV emerged in 2014 with the simultaneous introduction of both Asian and East/Central/South African (ECSA) genotypes. Laboratorial diagnosis of CHIKV is mainly performed by molecular and serological assays, with the latter more widely used. Although many commercial kits are available, their costs are still high for many underdeveloped and developing countries where the virus circulates. Here we described the development and evaluation of a multi-epitope recombinant protein-based IgG-ELISA (MULTREC IgG-ELISA) test for the specific detection of anti-CHIKV antibodies in clinical samples, as an alternative approach for laboratorial diagnosis. The MULTREC IgG-ELISA showed 86.36% of sensitivity and 100% of specificity, and no cross-reactivity with other exanthematic diseases was observed. The recombinant protein was expressed from the binary system insect cell/baculovirus using the crystal-forming baculoviral protein polyhedrin as a carrier of the target recombinant protein to facilitate recovery. The crystals were at least 10 times smaller in size and had an amorphous shape when compared to the polyhedrin wild-type crystal. The assay uses a multi-epitope antigen, representing two replicates of 18 amino acid sequences from the E2 region and a sequence of 17 amino acids from the nsP3 region of CHIKV. The recombinant protein was highly expressed, easy to purify and has demonstrated its usefulness in confirming chikungunya exposure, indeed showing a good potential tool for epidemiological surveillance.
    Language English
    Publishing date 2022-07-18
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2720891-6
    ISSN 2076-2607
    ISSN 2076-2607
    DOI 10.3390/microorganisms10071451
    Database MEDical Literature Analysis and Retrieval System OnLINE

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