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...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2016-05-01
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| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | http://europepmc.org/articles/PMC4861341?pdf=render |
| _version_ | 1818495160054448128 |
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| 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 |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-10T18:16:41Z |
| format | Article |
| id | doaj.art-baef6345a0f64c309edcc88120035c00 |
| institution | Directory Open Access Journal |
| issn | 1935-2727 1935-2735 |
| language | English |
| last_indexed | 2024-12-10T18:16:41Z |
| publishDate | 2016-05-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj.art-baef6345a0f64c309edcc88120035c002022-12-22T01:38:19ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352016-05-01105e000470410.1371/journal.pntd.0004704Using Rapid Diagnostic Tests as a Source of Viral RNA for Dengue Serotyping by RT-PCR - A Novel Epidemiological Tool.Manivanh VongsouvathKoukeo PhommasoneOnanong SengvilaipaseuthNathamon KosoltanapiwatNarisara ChantratitaStuart D BlacksellSue J LeeXavier de LamballerieMayfong MayxaySommay KeomanyPaul N NewtonAudrey Dubot-PérèsBACKGROUND: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.http://europepmc.org/articles/PMC4861341?pdf=render |
| spellingShingle | 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. PLoS Neglected Tropical Diseases |
| title | 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_short | 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 |
| url | http://europepmc.org/articles/PMC4861341?pdf=render |
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