Dataset Information

Using Rapid Diagnostic Tests as a Source of Viral RNA for Dengue Serotyping by RT-PCR - A Novel Epidemiological Tool.

ABSTRACT:

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.

SUBMITTER: Vongsouvath M

PROVIDER: S-EPMC4861341 | biostudies-literature | 2016 May

REPOSITORIES: biostudies-literature

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Publications

Using Rapid Diagnostic Tests as a Source of Viral RNA for Dengue Serotyping by RT-PCR - A Novel Epidemiological Tool.

Vongsouvath Manivanh M Phommasone Koukeo K Sengvilaipaseuth Onanong O Kosoltanapiwat Nathamon N Chantratita Narisara N Blacksell Stuart D SD Lee Sue J SJ de Lamballerie Xavier X Mayxay Mayfong M Keomany Sommay S Newton Paul N PN Dubot-Pérès Audrey A

PLoS neglected tropical diseases 20160509 5

<h4>Background</h4>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 t ...[more]

PMID: 27159058

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Project description:Four antigenically different dengue virus serotypes (DENV-1, DENV-2, DENV-3 and DENV-4) are known to cause infections in humans. Some of these are known to cause more severe disease than the others. Chances for developing Dengue hemorrhagic fever-dengue shock syndrome (DHF-DSS) increases significantly with history of previous infection with one of the four serotypes. Therefore, early diagnosis, serotyping and providing early warning of dengue fever epidemics to concerned authorities becomes very important for better patient outcome and to curb the rapid spread in the community. During the 2014 outbreak, a total of 100 samples from suspected cases of dengue were collected. NS1 antigen based rapid test was used for serological diagnosis. Dengue complex one step reverse transcription-polymerase chain reaction was performed to look for presence of viral RNA. Single tube multiplex RT-PCR was also performed to look for infecting serotype. CDC Dengue Multiplex Real Time PCR assay was performed for rapid diagnosis and simultaneous serotyping of the dengue virus. Out of the 100 samples screened, 69 were found to be positive by NS1Ag Rapid test. 34 samples were found positive by dengue consensus RT-PCR assay. 22 samples were found to be positive by single tube Dengue multiplex RT-PCR assay. Serotype DEN-2 was present in maximum numbers followed by DEN-3. 44 samples were found positive by DENV CDC Multiplex Real time PCR assay. DEN-2 was found in maximum numbers followed by DEN-1. Dengue remains to be an important health problem in India and across the globe. Few serotypes of dengue are more dangerous than the others. Rapid diagnosis and serotyping remains the key for better patient management and prevention of disease spreading in the community. Highly sensitive, specific and rapid CDC real time RT-PCR assay was found to be most promising tool among all available molecular diagnostic methods. This will serve a rapid and reliable simultaneous dengue virus detection as well serotyping assay in near future for rapid identification of dengue suspected sample screening.

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Project description:The coronavirus disease 2019 (COVID-19) cases could be symptomatic or asymptomatic. We (1) characterized and analyzed data collected from the first cohort of reverse transcriptase polymerase chain reaction (RT-PCR)-confirmed COVID-19 cases reported in the Emirate of Abu Dhabi, United Arab Emirates, according to the symptomatic state, and (2) identified factors associated with the symptomatic state. The association between the symptomatic state and testing positive in three subsequent RT-PCR testing rounds was also quantified. Between February 28 and April 8, 2020, 1,249 cases were reported. Sociodemographic characteristics, working status, travel history, and chronic comorbidities of 791 cases were analyzed according to the symptomatic state (symptomatic or asymptomatic). After the first confirmatory test, the results of three subsequent tests were analyzed. The mean age of the 791 cases was 35.6 ± 12.7 years (range: 1-81). Nearly 57.0% of cases were symptomatic. The two most frequent symptoms were fever (58.0%) and cough (41.0%). Symptomatic cases (mean age 36.3 ± 12.6 years) were significantly older than asymptomatic cases (mean age 34.5 ± 12.7 years). Compared with nonworking populations, working in public places (adjusted odds ratio (aOR), 1.76, 95% confidence interval (95% CI): 1.11-2.80), healthcare settings (aOR, 2.09, 95% CI: 1.01-4.31), or in the aviation and tourism sectors (aOR, 2.24, 95% CI: 1.14-4.40) was independently associated with the symptomatic state. Reporting at least one chronic comorbidity was also associated with symptomatic cases (aOR, 1.76, 95% CI: 1.03-3.01). Compared with asymptomatic cases, symptomatic cases had a prolonged duration of viral shedding and consistent odds of ?2 positive COVID-19 tests result out of the three subsequent testing rounds. A substantial proportion of the diagnosed COVID-19 cases in the Emirate of Abu Dhabi were asymptomatic. Quarantining asymptomatic cases, implementing prevention measures, and raising awareness among populations working in high-risk settings are warranted.

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Dataset Information

Using Rapid Diagnostic Tests as a Source of Viral RNA for Dengue Serotyping by RT-PCR - A Novel Epidemiological Tool.

Background

Methodology/principal findings

Conclusions/significance

Publications

Using Rapid Diagnostic Tests as a Source of Viral RNA for Dengue Serotyping by RT-PCR - A Novel Epidemiological Tool.

Similar Datasets

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