An ELISA-based method for detection of rabies virus nucleoprotein-specific antibodies in human antemortem samples.
Rabies is a fatal encephalitic disease in humans and animals caused by lyssaviruses, most commonly rabies virus (RABV). Human antemortem diagnosis of rabies is a complex process involving multiple sample types and tests for the detection of antibodies, antigen (protein), and nucleic acids (genomic R...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2018-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC6221316?pdf=render |
| _version_ | 1818319960866291712 |
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| author | Susan Realegeno Michael Niezgoda Pamela A Yager Amrita Kumar Laboni Hoque Lillian Orciari Suryaprakash Sambhara Victoria A Olson Panayampalli Subbian Satheshkumar |
| author_facet | Susan Realegeno Michael Niezgoda Pamela A Yager Amrita Kumar Laboni Hoque Lillian Orciari Suryaprakash Sambhara Victoria A Olson Panayampalli Subbian Satheshkumar |
| author_sort | Susan Realegeno |
| collection | DOAJ |
| description | Rabies is a fatal encephalitic disease in humans and animals caused by lyssaviruses, most commonly rabies virus (RABV). Human antemortem diagnosis of rabies is a complex process involving multiple sample types and tests for the detection of antibodies, antigen (protein), and nucleic acids (genomic RNA). Serological diagnosis of human rabies includes the detection of either neutralizing or binding antibodies in the cerebrospinal fluid (CSF) or serum samples from unimmunized individuals without prior rabies vaccination or passive immunization with purified immunoglobulins. While neutralizing antibodies are targeted against the surface-expressed glycoprotein (G protein), binding antibodies to viral antigens are predominantly against the nucleoprotein (N protein), although there can be antibodies against all RABV-expressed proteins. To determine N protein-specific antibody responses in the CSF and serum during RABV infection, we developed an enzyme-linked immunosorbent assay (ELISA) with purified recombinant N protein expressed in E. coli. N protein-specific immunoglobulin (Ig) subtypes IgG and IgM were detected in the CSF or serum of previously diagnosed human rabies cases. In addition, anti-N protein seroconversion was demonstrated over the course of illness in individual rabies cases. We compared the N protein ELISA results to those of an indirect fluorescent antibody (IFA) test, the current binding antibody assay used in diagnosis, and show that our ELISA is consistent with the IFA test. Sensitivity and specificity of the N protein ELISA ranged from 78.38-100% and 75.76-96.77% with respect to the IFA results. Our data provide evidence for the use of an N protein ELISA as an additional option for the detection of RABV-specific IgG or IgM antibodies in human CSF or serum specimens. |
| first_indexed | 2024-12-13T10:17:25Z |
| format | Article |
| id | doaj.art-3f8b0bfd72b4452d951dc2907a142a07 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-13T10:17:25Z |
| publishDate | 2018-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-3f8b0bfd72b4452d951dc2907a142a072022-12-21T23:51:16ZengPublic Library of Science (PLoS)PLoS ONE1932-62032018-01-011311e020700910.1371/journal.pone.0207009An ELISA-based method for detection of rabies virus nucleoprotein-specific antibodies in human antemortem samples.Susan RealegenoMichael NiezgodaPamela A YagerAmrita KumarLaboni HoqueLillian OrciariSuryaprakash SambharaVictoria A OlsonPanayampalli Subbian SatheshkumarRabies is a fatal encephalitic disease in humans and animals caused by lyssaviruses, most commonly rabies virus (RABV). Human antemortem diagnosis of rabies is a complex process involving multiple sample types and tests for the detection of antibodies, antigen (protein), and nucleic acids (genomic RNA). Serological diagnosis of human rabies includes the detection of either neutralizing or binding antibodies in the cerebrospinal fluid (CSF) or serum samples from unimmunized individuals without prior rabies vaccination or passive immunization with purified immunoglobulins. While neutralizing antibodies are targeted against the surface-expressed glycoprotein (G protein), binding antibodies to viral antigens are predominantly against the nucleoprotein (N protein), although there can be antibodies against all RABV-expressed proteins. To determine N protein-specific antibody responses in the CSF and serum during RABV infection, we developed an enzyme-linked immunosorbent assay (ELISA) with purified recombinant N protein expressed in E. coli. N protein-specific immunoglobulin (Ig) subtypes IgG and IgM were detected in the CSF or serum of previously diagnosed human rabies cases. In addition, anti-N protein seroconversion was demonstrated over the course of illness in individual rabies cases. We compared the N protein ELISA results to those of an indirect fluorescent antibody (IFA) test, the current binding antibody assay used in diagnosis, and show that our ELISA is consistent with the IFA test. Sensitivity and specificity of the N protein ELISA ranged from 78.38-100% and 75.76-96.77% with respect to the IFA results. Our data provide evidence for the use of an N protein ELISA as an additional option for the detection of RABV-specific IgG or IgM antibodies in human CSF or serum specimens.http://europepmc.org/articles/PMC6221316?pdf=render |
| spellingShingle | Susan Realegeno Michael Niezgoda Pamela A Yager Amrita Kumar Laboni Hoque Lillian Orciari Suryaprakash Sambhara Victoria A Olson Panayampalli Subbian Satheshkumar An ELISA-based method for detection of rabies virus nucleoprotein-specific antibodies in human antemortem samples. PLoS ONE |
| title | An ELISA-based method for detection of rabies virus nucleoprotein-specific antibodies in human antemortem samples. |
| title_full | An ELISA-based method for detection of rabies virus nucleoprotein-specific antibodies in human antemortem samples. |
| title_fullStr | An ELISA-based method for detection of rabies virus nucleoprotein-specific antibodies in human antemortem samples. |
| title_full_unstemmed | An ELISA-based method for detection of rabies virus nucleoprotein-specific antibodies in human antemortem samples. |
| title_short | An ELISA-based method for detection of rabies virus nucleoprotein-specific antibodies in human antemortem samples. |
| title_sort | elisa based method for detection of rabies virus nucleoprotein specific antibodies in human antemortem samples |
| url | http://europepmc.org/articles/PMC6221316?pdf=render |
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