Preliminary multiplex microarray IgG immunoassay for the diagnosis of toxoplasmosis and rubella

Preliminary multiplex microarray IgG immunoassay for the diagnosis of toxoplasmosis and rubella

BACKGROUND During pregnancy, toxoplasmosis and rubella can cause serious damage to the mother and the foetus through vertical transmission. Early diagnosis enables implementation of health measures aimed at preventing vertical transmission and minimising damage caused by these diseases. OBJECTIVE Here, we report the development of a multiplex assay for simultaneous detection of IgG antibodies produced during toxoplasmosis and rubella infection. METHODS This assay is based on xMap technology. Initially, by singleplex assays, we evaluated the following antigens: one Toxoplasma gondii lysate; two antigenic extracts of T. gondii (TOX8131 and TOX8122); fragments of T. gondii antigens [SAG-1 (amino acids 45-198), GRA-7 (24-100), GRA-1 (57-149), ROP-4, and MIC-3 (234-306)]; two chimeric antigens composed of fragments of SAG-1, GRA-7, and P35 (CTOX and CTOXH); and fragments of Rubella virus antigens [E-1 (157-176, 213-239, 374-390), E-2 (31-105), and C (1-123)]. FINDINGS A multiplex assay to simultaneously diagnose toxoplasmosis and rubella was designed with the best-performing antigens in singleplex and multiplex assays, which included CTOXH, T. gondii lysate, TOX8131, E-1, and E-2. The multiplex assay showed 100% sensitivity and specificity for anti-T. gondii IgG detection and 95.6% sensitivity and 100% specificity for anti-R. virus IgG detection. MAIN CONCLUSIONS We found that, despite the difficulties related to developing multiplex systems, different types of antigens (extracts and recombinant proteins) can be used to develop high-performance diagnostic tests. The assay developed is suitable to screen for prior T. gondii and R. virus infections, because it is a rapid, high-throughput, low-cost alternative to the current standard diagnostic tools, which require multiple individual tests.

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Main Authors: Baschirotto,Priscila T, Krieger,Marco A, Foti,Leonardo
Format: Digital revista
Language:English
Published: Instituto Oswaldo Cruz, Ministério da Saúde 2017
Online Access:http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762017000600428
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spelling oai:scielo:S0074-027620170006004282017-05-26Preliminary multiplex microarray IgG immunoassay for the diagnosis of toxoplasmosis and rubellaBaschirotto,Priscila TKrieger,Marco AFoti,Leonardo toxoplasmosis rubella diagnosis multiplex liquid microarray BACKGROUND During pregnancy, toxoplasmosis and rubella can cause serious damage to the mother and the foetus through vertical transmission. Early diagnosis enables implementation of health measures aimed at preventing vertical transmission and minimising damage caused by these diseases. OBJECTIVE Here, we report the development of a multiplex assay for simultaneous detection of IgG antibodies produced during toxoplasmosis and rubella infection. METHODS This assay is based on xMap technology. Initially, by singleplex assays, we evaluated the following antigens: one Toxoplasma gondii lysate; two antigenic extracts of T. gondii (TOX8131 and TOX8122); fragments of T. gondii antigens [SAG-1 (amino acids 45-198), GRA-7 (24-100), GRA-1 (57-149), ROP-4, and MIC-3 (234-306)]; two chimeric antigens composed of fragments of SAG-1, GRA-7, and P35 (CTOX and CTOXH); and fragments of Rubella virus antigens [E-1 (157-176, 213-239, 374-390), E-2 (31-105), and C (1-123)]. FINDINGS A multiplex assay to simultaneously diagnose toxoplasmosis and rubella was designed with the best-performing antigens in singleplex and multiplex assays, which included CTOXH, T. gondii lysate, TOX8131, E-1, and E-2. The multiplex assay showed 100% sensitivity and specificity for anti-T. gondii IgG detection and 95.6% sensitivity and 100% specificity for anti-R. virus IgG detection. MAIN CONCLUSIONS We found that, despite the difficulties related to developing multiplex systems, different types of antigens (extracts and recombinant proteins) can be used to develop high-performance diagnostic tests. The assay developed is suitable to screen for prior T. gondii and R. virus infections, because it is a rapid, high-throughput, low-cost alternative to the current standard diagnostic tools, which require multiple individual tests.info:eu-repo/semantics/openAccessInstituto Oswaldo Cruz, Ministério da SaúdeMemórias do Instituto Oswaldo Cruz v.112 n.6 20172017-06-01info:eu-repo/semantics/articletext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762017000600428en10.1590/0074-02760160509
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countrycode BR
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libraryname SciELO
language English
format Digital
author Baschirotto,Priscila T
Krieger,Marco A
Foti,Leonardo
spellingShingle Baschirotto,Priscila T
Krieger,Marco A
Foti,Leonardo
Preliminary multiplex microarray IgG immunoassay for the diagnosis of toxoplasmosis and rubella
author_facet Baschirotto,Priscila T
Krieger,Marco A
Foti,Leonardo
author_sort Baschirotto,Priscila T
title Preliminary multiplex microarray IgG immunoassay for the diagnosis of toxoplasmosis and rubella
title_short Preliminary multiplex microarray IgG immunoassay for the diagnosis of toxoplasmosis and rubella
title_full Preliminary multiplex microarray IgG immunoassay for the diagnosis of toxoplasmosis and rubella
title_fullStr Preliminary multiplex microarray IgG immunoassay for the diagnosis of toxoplasmosis and rubella
title_full_unstemmed Preliminary multiplex microarray IgG immunoassay for the diagnosis of toxoplasmosis and rubella
title_sort preliminary multiplex microarray igg immunoassay for the diagnosis of toxoplasmosis and rubella
description BACKGROUND During pregnancy, toxoplasmosis and rubella can cause serious damage to the mother and the foetus through vertical transmission. Early diagnosis enables implementation of health measures aimed at preventing vertical transmission and minimising damage caused by these diseases. OBJECTIVE Here, we report the development of a multiplex assay for simultaneous detection of IgG antibodies produced during toxoplasmosis and rubella infection. METHODS This assay is based on xMap technology. Initially, by singleplex assays, we evaluated the following antigens: one Toxoplasma gondii lysate; two antigenic extracts of T. gondii (TOX8131 and TOX8122); fragments of T. gondii antigens [SAG-1 (amino acids 45-198), GRA-7 (24-100), GRA-1 (57-149), ROP-4, and MIC-3 (234-306)]; two chimeric antigens composed of fragments of SAG-1, GRA-7, and P35 (CTOX and CTOXH); and fragments of Rubella virus antigens [E-1 (157-176, 213-239, 374-390), E-2 (31-105), and C (1-123)]. FINDINGS A multiplex assay to simultaneously diagnose toxoplasmosis and rubella was designed with the best-performing antigens in singleplex and multiplex assays, which included CTOXH, T. gondii lysate, TOX8131, E-1, and E-2. The multiplex assay showed 100% sensitivity and specificity for anti-T. gondii IgG detection and 95.6% sensitivity and 100% specificity for anti-R. virus IgG detection. MAIN CONCLUSIONS We found that, despite the difficulties related to developing multiplex systems, different types of antigens (extracts and recombinant proteins) can be used to develop high-performance diagnostic tests. The assay developed is suitable to screen for prior T. gondii and R. virus infections, because it is a rapid, high-throughput, low-cost alternative to the current standard diagnostic tools, which require multiple individual tests.
publisher Instituto Oswaldo Cruz, Ministério da Saúde
publishDate 2017
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762017000600428
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AT kriegermarcoa preliminarymultiplexmicroarrayiggimmunoassayforthediagnosisoftoxoplasmosisandrubella
AT fotileonardo preliminarymultiplexmicroarrayiggimmunoassayforthediagnosisoftoxoplasmosisandrubella
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