<i>Strongyloides</i>-Specific IgE Phage cDNA Clones and Development of a Novel ELISA for Strongyloidiasis
Strongyloidiasis, caused mainly by the nematode <i>Strongyloides stercoralis</i>, is prevalent worldwide and potentially fatal in immunosuppressed patients. We report on a new IgE biomarker to diagnose <i>Strongyloides</i> infection. Sera from two groups infected with <i&g...
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doaj-b3d0567edbff41c79bb4358ff2f40b7f2021-06-01T01:33:33ZengMDPI AGDiagnostics2075-44182021-05-011198598510.3390/diagnostics11060985<i>Strongyloides</i>-Specific IgE Phage cDNA Clones and Development of a Novel ELISA for StrongyloidiasisHussain Ahmad0Norsyahida Arifin1Thomas J. Nolan2James B. Lok3Nor Suhada Anuar4Rahmah Noordin5Institute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang 11800, MalaysiaInstitute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang 11800, MalaysiaDepartment of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, USADepartment of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA 19104, USAInstitute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang 11800, MalaysiaInstitute for Research in Molecular Medicine (INFORMM), Universiti Sains Malaysia, Penang 11800, MalaysiaStrongyloidiasis, caused mainly by the nematode <i>Strongyloides stercoralis</i>, is prevalent worldwide and potentially fatal in immunosuppressed patients. We report on a new IgE biomarker to diagnose <i>Strongyloides</i> infection. Sera from two groups infected with <i>Strongyloides</i> served as positive samples: Group 1A, in which infection was confirmed by stool-microscopy and/or stool-polymerase chain reaction (PCR) and was seropositive by an IgG-enzyme linked immunosorbent assay (ELISA) and an IgG4 rapid test, and Group 1B in which infection was confirmed by stool-PCR but was seronegative. Negative samples (controls) comprised infections with other parasites (Group II) and healthy donors (Group III). Immunoscreenings of an <i>S. stercoralis</i> complementary DNA (cDNA) library were performed, and the cDNA clone with the highest diagnostic potential (clone A133) was selected for recombinant protein production and then evaluated using IgE Western blot and ELISA. The Western blot showed that the recombinant protein (rA133) was 100% reactive with Group IA (<i>n </i>= 10) and Group IB (<i>n </i>= 5), and 96% non-reactive with Groups II and III (<i>n </i>= 25). Subsequently, the IgE-ELISA was developed and showed 100% diagnostic sensitivity in Groups IA (<i>n </i>= 32) and IB (<i>n </i>= 11); and 99.3% specificity in Groups II and III (<i>n </i>= 144). In conclusion, this study has identified rA133 as a novel recombinant protein with potential diagnostic value, and that the IgE-ELISA incorporating this protein may be useful for patient diagnosis and epidemiological studies.https://www.mdpi.com/2075-4418/11/6/985<i>Strongyloides stercoralis</i>strongyloidiasisimmunoscreeningcomplementary DNA (cDNA) libraryrecombinant antigenserodiagnosis |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Hussain Ahmad Norsyahida Arifin Thomas J. Nolan James B. Lok Nor Suhada Anuar Rahmah Noordin |
spellingShingle |
Hussain Ahmad Norsyahida Arifin Thomas J. Nolan James B. Lok Nor Suhada Anuar Rahmah Noordin <i>Strongyloides</i>-Specific IgE Phage cDNA Clones and Development of a Novel ELISA for Strongyloidiasis Diagnostics <i>Strongyloides stercoralis</i> strongyloidiasis immunoscreening complementary DNA (cDNA) library recombinant antigen serodiagnosis |
author_facet |
Hussain Ahmad Norsyahida Arifin Thomas J. Nolan James B. Lok Nor Suhada Anuar Rahmah Noordin |
author_sort |
Hussain Ahmad |
title |
<i>Strongyloides</i>-Specific IgE Phage cDNA Clones and Development of a Novel ELISA for Strongyloidiasis |
title_short |
<i>Strongyloides</i>-Specific IgE Phage cDNA Clones and Development of a Novel ELISA for Strongyloidiasis |
title_full |
<i>Strongyloides</i>-Specific IgE Phage cDNA Clones and Development of a Novel ELISA for Strongyloidiasis |
title_fullStr |
<i>Strongyloides</i>-Specific IgE Phage cDNA Clones and Development of a Novel ELISA for Strongyloidiasis |
title_full_unstemmed |
<i>Strongyloides</i>-Specific IgE Phage cDNA Clones and Development of a Novel ELISA for Strongyloidiasis |
title_sort |
<i>strongyloides</i>-specific ige phage cdna clones and development of a novel elisa for strongyloidiasis |
publisher |
MDPI AG |
series |
Diagnostics |
issn |
2075-4418 |
publishDate |
2021-05-01 |
description |
Strongyloidiasis, caused mainly by the nematode <i>Strongyloides stercoralis</i>, is prevalent worldwide and potentially fatal in immunosuppressed patients. We report on a new IgE biomarker to diagnose <i>Strongyloides</i> infection. Sera from two groups infected with <i>Strongyloides</i> served as positive samples: Group 1A, in which infection was confirmed by stool-microscopy and/or stool-polymerase chain reaction (PCR) and was seropositive by an IgG-enzyme linked immunosorbent assay (ELISA) and an IgG4 rapid test, and Group 1B in which infection was confirmed by stool-PCR but was seronegative. Negative samples (controls) comprised infections with other parasites (Group II) and healthy donors (Group III). Immunoscreenings of an <i>S. stercoralis</i> complementary DNA (cDNA) library were performed, and the cDNA clone with the highest diagnostic potential (clone A133) was selected for recombinant protein production and then evaluated using IgE Western blot and ELISA. The Western blot showed that the recombinant protein (rA133) was 100% reactive with Group IA (<i>n </i>= 10) and Group IB (<i>n </i>= 5), and 96% non-reactive with Groups II and III (<i>n </i>= 25). Subsequently, the IgE-ELISA was developed and showed 100% diagnostic sensitivity in Groups IA (<i>n </i>= 32) and IB (<i>n </i>= 11); and 99.3% specificity in Groups II and III (<i>n </i>= 144). In conclusion, this study has identified rA133 as a novel recombinant protein with potential diagnostic value, and that the IgE-ELISA incorporating this protein may be useful for patient diagnosis and epidemiological studies. |
topic |
<i>Strongyloides stercoralis</i> strongyloidiasis immunoscreening complementary DNA (cDNA) library recombinant antigen serodiagnosis |
url |
https://www.mdpi.com/2075-4418/11/6/985 |
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