Using Rapid Diagnostic Tests as a Source of Viral RNA for Dengue Serotyping by RT-PCR - A Novel Epidemiological Tool.
BACKGROUND:Dengue virus infection causes major public health problems in tropical and subtropical areas. In many endemic areas, including the Lao PDR, inadequate access to laboratory facilities is a major obstacle to surveillance and study of dengue epidemiology. Filter paper is widely used for bloo...
Published in: | PLOS Neglected Tropical Diseases |
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ftdoajarticles:oai:doaj.org/article:baef6345a0f64c309edcc88120035c00 2023-05-15T15:12:57+02:00 Using Rapid Diagnostic Tests as a Source of Viral RNA for Dengue Serotyping by RT-PCR - A Novel Epidemiological Tool. Manivanh Vongsouvath Koukeo Phommasone Onanong Sengvilaipaseuth Nathamon Kosoltanapiwat Narisara Chantratita Stuart D Blacksell Sue J Lee Xavier de Lamballerie Mayfong Mayxay Sommay Keomany Paul N Newton Audrey Dubot-Pérès 2016-05-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0004704 https://doaj.org/article/baef6345a0f64c309edcc88120035c00 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC4861341?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0004704 https://doaj.org/article/baef6345a0f64c309edcc88120035c00 PLoS Neglected Tropical Diseases, Vol 10, Iss 5, p e0004704 (2016) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2016 ftdoajarticles https://doi.org/10.1371/journal.pntd.0004704 2022-12-31T00:49:30Z BACKGROUND:Dengue virus infection causes major public health problems in tropical and subtropical areas. In many endemic areas, including the Lao PDR, inadequate access to laboratory facilities is a major obstacle to surveillance and study of dengue epidemiology. Filter paper is widely used for blood collection for subsequent laboratory testing for antibody and nucleic acid detection. For the first time, we demonstrate that dengue viral RNA can be extracted from dengue rapid diagnostic tests (RDT) and then submitted to real-time RT-PCR for serotyping. METHODOLOGY/PRINCIPAL FINDINGS:We evaluated the Standard Diagnostics (SD) Bioline Dengue Duo RDT, a commonly used test in dengue endemic areas. First, using the QIAamp RNA kit, dengue RNA was purified from the sample pad of the NS1 RDT loaded with virus isolates of the four serotypes, then quantified by RT-PCR. We observed greater recovery of virus, with a mean of 27 times more RNA recovered from RDT, than from filter paper. Second, we evaluated dengue NS1 RDTs from patients at Mahosot Hospital, Vientiane, (99 patients) and from rural Salavan Provincial Hospital (362 patients). There was good agreement between dengue RT-PCR from NS1 RDT with RT-PCR performed on RNA extracted from patient sera, either using RDT loaded with blood (82.8% and 91.4%, in Vientiane and Salavan, respectively) or serum (91.9% and 93.9%). There was 100% concordance between RDT and serum RT-PCR of infecting dengue serotype. CONCLUSIONS/SIGNIFICANCE:Therefore, the collection of NS1 positive RDTs, which do not require cold storage, may be a novel approach for dengue serotyping by RT-PCR and offers promising prospects for the collection of epidemiological data from previously inaccessible tropical areas to aid surveillance and public health interventions. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 10 5 e0004704 |
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 Manivanh Vongsouvath Koukeo Phommasone Onanong Sengvilaipaseuth Nathamon Kosoltanapiwat Narisara Chantratita Stuart D Blacksell Sue J Lee Xavier de Lamballerie Mayfong Mayxay Sommay Keomany Paul N Newton Audrey Dubot-Pérès Using Rapid Diagnostic Tests as a Source of Viral RNA for Dengue Serotyping by RT-PCR - A Novel Epidemiological Tool. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
BACKGROUND:Dengue virus infection causes major public health problems in tropical and subtropical areas. In many endemic areas, including the Lao PDR, inadequate access to laboratory facilities is a major obstacle to surveillance and study of dengue epidemiology. Filter paper is widely used for blood collection for subsequent laboratory testing for antibody and nucleic acid detection. For the first time, we demonstrate that dengue viral RNA can be extracted from dengue rapid diagnostic tests (RDT) and then submitted to real-time RT-PCR for serotyping. METHODOLOGY/PRINCIPAL FINDINGS:We evaluated the Standard Diagnostics (SD) Bioline Dengue Duo RDT, a commonly used test in dengue endemic areas. First, using the QIAamp RNA kit, dengue RNA was purified from the sample pad of the NS1 RDT loaded with virus isolates of the four serotypes, then quantified by RT-PCR. We observed greater recovery of virus, with a mean of 27 times more RNA recovered from RDT, than from filter paper. Second, we evaluated dengue NS1 RDTs from patients at Mahosot Hospital, Vientiane, (99 patients) and from rural Salavan Provincial Hospital (362 patients). There was good agreement between dengue RT-PCR from NS1 RDT with RT-PCR performed on RNA extracted from patient sera, either using RDT loaded with blood (82.8% and 91.4%, in Vientiane and Salavan, respectively) or serum (91.9% and 93.9%). There was 100% concordance between RDT and serum RT-PCR of infecting dengue serotype. CONCLUSIONS/SIGNIFICANCE:Therefore, the collection of NS1 positive RDTs, which do not require cold storage, may be a novel approach for dengue serotyping by RT-PCR and offers promising prospects for the collection of epidemiological data from previously inaccessible tropical areas to aid surveillance and public health interventions. |
format |
Article in Journal/Newspaper |
author |
Manivanh Vongsouvath Koukeo Phommasone Onanong Sengvilaipaseuth Nathamon Kosoltanapiwat Narisara Chantratita Stuart D Blacksell Sue J Lee Xavier de Lamballerie Mayfong Mayxay Sommay Keomany Paul N Newton Audrey Dubot-Pérès |
author_facet |
Manivanh Vongsouvath Koukeo Phommasone Onanong Sengvilaipaseuth Nathamon Kosoltanapiwat Narisara Chantratita Stuart D Blacksell Sue J Lee Xavier de Lamballerie Mayfong Mayxay Sommay Keomany Paul N Newton Audrey Dubot-Pérès |
author_sort |
Manivanh Vongsouvath |
title |
Using Rapid Diagnostic Tests as a Source of Viral RNA for Dengue Serotyping by RT-PCR - A Novel Epidemiological Tool. |
title_short |
Using Rapid Diagnostic Tests as a Source of Viral RNA for Dengue Serotyping by RT-PCR - A Novel Epidemiological Tool. |
title_full |
Using Rapid Diagnostic Tests as a Source of Viral RNA for Dengue Serotyping by RT-PCR - A Novel Epidemiological Tool. |
title_fullStr |
Using Rapid Diagnostic Tests as a Source of Viral RNA for Dengue Serotyping by RT-PCR - A Novel Epidemiological Tool. |
title_full_unstemmed |
Using Rapid Diagnostic Tests as a Source of Viral RNA for Dengue Serotyping by RT-PCR - A Novel Epidemiological Tool. |
title_sort |
using rapid diagnostic tests as a source of viral rna for dengue serotyping by rt-pcr - a novel epidemiological tool. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2016 |
url |
https://doi.org/10.1371/journal.pntd.0004704 https://doaj.org/article/baef6345a0f64c309edcc88120035c00 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 10, Iss 5, p e0004704 (2016) |
op_relation |
http://europepmc.org/articles/PMC4861341?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0004704 https://doaj.org/article/baef6345a0f64c309edcc88120035c00 |
op_doi |
https://doi.org/10.1371/journal.pntd.0004704 |
container_title |
PLOS Neglected Tropical Diseases |
container_volume |
10 |
container_issue |
5 |
container_start_page |
e0004704 |
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1766343566465957888 |