Detection of toxoplasmic encephalitis in HIV positive patients in urine with hydrogel nanoparticles.
Background Diagnosis of toxoplasmic encephalitis (TE) is challenging under the best clinical circumstances. The poor clinical sensitivity of quantitative polymerase chain reaction (qPCR) for Toxoplasma in blood and CSF and the limited availability of molecular diagnostics and imaging technology leav...
Published in: | PLOS Neglected Tropical Diseases |
---|---|
Main Authors: | , , , , , , , , , , , , , , , , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
Public Library of Science (PLoS)
2021
|
Subjects: | |
Online Access: | https://doi.org/10.1371/journal.pntd.0009199 https://doaj.org/article/5bb57224791f4f32be819732c3f0343a |
id |
ftdoajarticles:oai:doaj.org/article:5bb57224791f4f32be819732c3f0343a |
---|---|
record_format |
openpolar |
spelling |
ftdoajarticles:oai:doaj.org/article:5bb57224791f4f32be819732c3f0343a 2023-05-15T15:15:35+02:00 Detection of toxoplasmic encephalitis in HIV positive patients in urine with hydrogel nanoparticles. Hannah E Steinberg Natalie M Bowman Andrea Diestra Cusi Ferradas Paul Russo Daniel E Clark Deanna Zhu Ruben Magni Edith Malaga Monica Diaz Viviana Pinedo-Cancino Cesar Ramal Asayag Maritza Calderón Vern B Carruthers Lance A Liotta Robert H Gilman Alessandra Luchini Toxoplasmosis working group in Peru and Bolivia 2021-03-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0009199 https://doaj.org/article/5bb57224791f4f32be819732c3f0343a EN eng Public Library of Science (PLoS) https://doi.org/10.1371/journal.pntd.0009199 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0009199 https://doaj.org/article/5bb57224791f4f32be819732c3f0343a PLoS Neglected Tropical Diseases, Vol 15, Iss 3, p e0009199 (2021) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2021 ftdoajarticles https://doi.org/10.1371/journal.pntd.0009199 2022-12-31T02:54:22Z Background Diagnosis of toxoplasmic encephalitis (TE) is challenging under the best clinical circumstances. The poor clinical sensitivity of quantitative polymerase chain reaction (qPCR) for Toxoplasma in blood and CSF and the limited availability of molecular diagnostics and imaging technology leaves clinicians in resource-limited settings with few options other than empiric treatment. Methology/principle findings Here we describe proof of concept for a novel urine diagnostics for TE using Poly-N-Isopropylacrylamide nanoparticles dyed with Reactive Blue-221 to concentrate antigens, substantially increasing the limit of detection. After nanoparticle-concentration, a standard western blotting technique with a monoclonal antibody was used for antigen detection. Limit of detection was 7.8pg/ml and 31.3pg/ml of T. gondii antigens GRA1 and SAG1, respectively. To characterize this diagnostic approach, 164 hospitalized HIV-infected patients with neurological symptoms compatible with TE were tested for 1) T. gondii serology (121/147, positive samples/total samples tested), 2) qPCR in cerebrospinal fluid (11/41), 3) qPCR in blood (10/112), and 4) urinary GRA1 (30/164) and SAG1 (12/164). GRA1 appears to be superior to SAG1 for detection of TE antigens in urine. Fifty-one HIV-infected, T. gondii seropositive but asymptomatic persons all tested negative by nanoparticle western blot and blood qPCR, suggesting the test has good specificity for TE for both GRA1 and SAG1. In a subgroup of 44 patients, urine samples were assayed with mass spectrometry parallel-reaction-monitoring (PRM) for the presence of T. gondii antigens. PRM identified antigens in 8 samples, 6 of which were concordant with the urine diagnostic. Conclusion/significances Our results demonstrate nanoparticle technology's potential for a noninvasive diagnostic test for TE. Moving forward, GRA1 is a promising target for antigen based diagnostics for TE. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 15 3 e0009199 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Hannah E Steinberg Natalie M Bowman Andrea Diestra Cusi Ferradas Paul Russo Daniel E Clark Deanna Zhu Ruben Magni Edith Malaga Monica Diaz Viviana Pinedo-Cancino Cesar Ramal Asayag Maritza Calderón Vern B Carruthers Lance A Liotta Robert H Gilman Alessandra Luchini Toxoplasmosis working group in Peru and Bolivia Detection of toxoplasmic encephalitis in HIV positive patients in urine with hydrogel nanoparticles. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Background Diagnosis of toxoplasmic encephalitis (TE) is challenging under the best clinical circumstances. The poor clinical sensitivity of quantitative polymerase chain reaction (qPCR) for Toxoplasma in blood and CSF and the limited availability of molecular diagnostics and imaging technology leaves clinicians in resource-limited settings with few options other than empiric treatment. Methology/principle findings Here we describe proof of concept for a novel urine diagnostics for TE using Poly-N-Isopropylacrylamide nanoparticles dyed with Reactive Blue-221 to concentrate antigens, substantially increasing the limit of detection. After nanoparticle-concentration, a standard western blotting technique with a monoclonal antibody was used for antigen detection. Limit of detection was 7.8pg/ml and 31.3pg/ml of T. gondii antigens GRA1 and SAG1, respectively. To characterize this diagnostic approach, 164 hospitalized HIV-infected patients with neurological symptoms compatible with TE were tested for 1) T. gondii serology (121/147, positive samples/total samples tested), 2) qPCR in cerebrospinal fluid (11/41), 3) qPCR in blood (10/112), and 4) urinary GRA1 (30/164) and SAG1 (12/164). GRA1 appears to be superior to SAG1 for detection of TE antigens in urine. Fifty-one HIV-infected, T. gondii seropositive but asymptomatic persons all tested negative by nanoparticle western blot and blood qPCR, suggesting the test has good specificity for TE for both GRA1 and SAG1. In a subgroup of 44 patients, urine samples were assayed with mass spectrometry parallel-reaction-monitoring (PRM) for the presence of T. gondii antigens. PRM identified antigens in 8 samples, 6 of which were concordant with the urine diagnostic. Conclusion/significances Our results demonstrate nanoparticle technology's potential for a noninvasive diagnostic test for TE. Moving forward, GRA1 is a promising target for antigen based diagnostics for TE. |
format |
Article in Journal/Newspaper |
author |
Hannah E Steinberg Natalie M Bowman Andrea Diestra Cusi Ferradas Paul Russo Daniel E Clark Deanna Zhu Ruben Magni Edith Malaga Monica Diaz Viviana Pinedo-Cancino Cesar Ramal Asayag Maritza Calderón Vern B Carruthers Lance A Liotta Robert H Gilman Alessandra Luchini Toxoplasmosis working group in Peru and Bolivia |
author_facet |
Hannah E Steinberg Natalie M Bowman Andrea Diestra Cusi Ferradas Paul Russo Daniel E Clark Deanna Zhu Ruben Magni Edith Malaga Monica Diaz Viviana Pinedo-Cancino Cesar Ramal Asayag Maritza Calderón Vern B Carruthers Lance A Liotta Robert H Gilman Alessandra Luchini Toxoplasmosis working group in Peru and Bolivia |
author_sort |
Hannah E Steinberg |
title |
Detection of toxoplasmic encephalitis in HIV positive patients in urine with hydrogel nanoparticles. |
title_short |
Detection of toxoplasmic encephalitis in HIV positive patients in urine with hydrogel nanoparticles. |
title_full |
Detection of toxoplasmic encephalitis in HIV positive patients in urine with hydrogel nanoparticles. |
title_fullStr |
Detection of toxoplasmic encephalitis in HIV positive patients in urine with hydrogel nanoparticles. |
title_full_unstemmed |
Detection of toxoplasmic encephalitis in HIV positive patients in urine with hydrogel nanoparticles. |
title_sort |
detection of toxoplasmic encephalitis in hiv positive patients in urine with hydrogel nanoparticles. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2021 |
url |
https://doi.org/10.1371/journal.pntd.0009199 https://doaj.org/article/5bb57224791f4f32be819732c3f0343a |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 15, Iss 3, p e0009199 (2021) |
op_relation |
https://doi.org/10.1371/journal.pntd.0009199 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0009199 https://doaj.org/article/5bb57224791f4f32be819732c3f0343a |
op_doi |
https://doi.org/10.1371/journal.pntd.0009199 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
15 |
container_issue |
3 |
container_start_page |
e0009199 |
_version_ |
1766345954248622080 |