Using rapid diagnostic tests as a source of viral RNA for dengue serotyping by RT-PCR : a novel epidemiological tool
| Autor: | Paul N. Newton, Onanong Sengvilaipaseuth, Narisara Chantratita, Audrey Dubot-Pérès, Koukeo Phommasone, Xavier de Lamballerie, Sommay Keomany, Stuart D. Blacksell, Sue J. Lee, Nathamon Kosoltanapiwat, Mayfong Mayxay, Manivanh Vongsouvath |
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| Přispěvatelé: | Wills, B |
| Rok vydání: | 2016 |
| Předmět: |
RNA viruses
0301 basic medicine Serotype Time Factors Physiology Artificial Gene Amplification and Extension Dengue virus Pathology and Laboratory Medicine medicine.disease_cause Polymerase Chain Reaction Chromatography Affinity Dengue fever Dengue 0302 clinical medicine Filter Paper Chlorocebus aethiops Epidemiology Medicine and Health Sciences Reverse Transcriptase Polymerase Chain Reaction lcsh:Public aspects of medicine Hematology Body Fluids 3. Good health Laboratory Equipment RNA isolation Blood Infectious Diseases Real-time polymerase chain reaction Vietnam Medical Microbiology Viral Pathogens Viruses Engineering and Technology RNA Viral RNA extraction Anatomy Pathogens Research Article medicine.medical_specialty lcsh:Arctic medicine. Tropical medicine lcsh:RC955-962 030106 microbiology 030231 tropical medicine Equipment Cold storage Biology Biomolecular isolation Microbiology Virus 03 medical and health sciences Extraction techniques parasitic diseases medicine Animals Humans Serotyping Molecular Biology Techniques Molecular Biology Microbial Pathogens Vero Cells Flaviviruses Organisms Public Health Environmental and Occupational Health Biology and Life Sciences lcsh:RA1-1270 Reverse Transcriptase-Polymerase Chain Reaction Dengue Virus medicine.disease Virology Research and analysis methods Reagent Kits Diagnostic |
| Zdroj: | PLoS Neglected Tropical Diseases, Vol 10, Iss 5, p e0004704 (2016) PLoS Neglected Tropical Diseases |
| Popis: | 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. Author Summary Dengue fever, caused by a virus transmitted by mosquitoes, is a public health problem in tropical and subtropical regions. Dengue Rapid Diagnostic Tests, in which a drop of blood is loaded onto a paper strip in a plastic cassette, are simple to use and have good diagnostic accuracy. They are becoming the test of choice for the management of dengue epidemics, especially in rural areas without laboratory facilities. However, four types of dengue virus circulate in most tropical areas and their patterns of circulation are of epidemiological importance since they play a role in the severity and propagation of the disease. We show, for the first time, that molecular amplification permitting dengue virus detection and typing can be performed directly from used positive RDTs, that can easily be transported. This novel approach has promising prospects for the collection of dengue epidemiological data from previously inaccessible tropical areas to aid surveillance and public health interventions. |
| Databáze: | OpenAIRE |
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