Real-time RT-PCR for the detection and quantitation of Oropouche virus

Rojas, Alejandra; Stittleburg, Victoria; Cardozo, Fátima; Bopp, Nathen E.; Cantero, César; López, Sanny; Bernal, Cynthia; Mendoza, Laura; Aguilar, Patricia V.; Pinsky, Benjamin A.; Guillén, Yvalena; Páez, Malvina; Waggoner, Jesse J.

doi:10.1016/j.diagmicrobio.2019.114894

Cited by 16 publications

(16 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Positive and negative controls were included on each run, and rRT-PCRs were analyzed and interpreted as previously described. 42-45…”

Section: Methodsmentioning

confidence: 99%

“…Eluates from optimization and analytical evaluation experiments were tested in a single-reaction multiplex rRT-PCR for ZIKV, CHIKV, and DENV or a singleplex rRT-PCR for OROV, both performed as previously described. 44, 45 For the clinical evaluation, eluates were tested in the DENV multiplex rRT-PCR, which is a serotype-specific assay for DENV detection and quantitation. 42, 43 All rRT-PCR reactions were performed in 20μL reactions of the SuperScript III Platinum One-Step qRT-PCR kit (Thermo Fisher) containing 5μL of eluate and run on a Rotor-Gene Q instrument (Qiagen).…”

Section: Methodsmentioning

confidence: 99%

“…Positive and negative controls were included on each run, and rRT-PCRs were analyzed and interpreted as previously described. [42][43][44][45] Statistics. Basic statistical analyses were performed in Excel (Microsoft, Redmond, WA).…”

Section: Rrt-pcrmentioning

confidence: 99%

See 2 more Smart Citations

Simple and economical RNA extraction and storage packets for viral detection from serum or plasma

Hernandez

Cardozo

Myers

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

RNA extraction is an essential step for detection and surveillance of common viral pathogens. Currently, sample processing and RNA extraction are costly and rely on proprietary materials that are difficult to acquire, maintain, and safely discard in low-resource settings. We developed an economical RNA extraction and storage protocol that eliminates the use of instrumentation, expensive materials, and cold chain requirements. Through an iterative process, we optimized viral lysis and RNA binding to and elution from glass fiber membranes. Protocol changes were evaluated by testing eluates in virus-specific real-time RT-PCRs (rRT-PCRs). Efficient, non-toxic viral lysis was achieved with a sucrose buffer including KCl, proteinase K and carrier RNA. Glass fiber membranes demonstrated concentration-dependent RNA binding of three arthropod-borne RNA viruses (arboviruses): dengue, chikungunya and Oropouche. Membrane binding was significantly increased in an acidic arginine binding buffer. For the clinical evaluation, 36 dengue virus (DENV)-positive serum samples were extracted in duplicate in the optimized protocol and results were compared to a commercial method. DENV RNA was successfully extracted from 71/72 replicates (98.6%) in the extraction packets, and rRT-PCR Ct values correlated between the techniques. Five clinical samples were selected to evaluate ambient-temperature storage up to 7 days on dried glass fiber membranes. DENV RNA was stable at 1, 3 and 7 days post extraction, with a mean difference in eluate RNA concentration of 0.14 log10 copies/uL. At a cost of $0.08 /sample, RNA extraction and storage packets address key limitations to available protocols and may increase capacity for molecular detection of RNA viruses. Keywords: ribonucleic acid, extraction, dengue virus, molecular testing

show abstract

“…Positive and negative controls were included on each run, and rRT-PCRs were analyzed and interpreted as previously described. 42-45…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Simple and economical RNA extraction and storage packets for viral detection from serum or plasma

Hernandez

Cardozo

Myers

et al. 2022

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Across all 56 articles included for OROV surveillance, there were eight separate countries where these investigations took place (Figure 3). There were 37 studies performed in Brazil (63%) [2,5,7,16,17,24,32,, followed by 11 studies in Peru (18.6%) [6,18,34,[68][69][70][71][72][73][74][75], with the remaining (18.6%) taking place in Ecuador [3,8,15,70,76], Paraguay [70,77], Suriname [78], Trinidad [4], Bolivia [70], and Costa Rica [79].…”

Section: Orov Surveillancementioning

confidence: 99%

“…,15,34,47,49,53-55,60,61,66,72,73,76], or both (13 studies)[4,7,16,17,24,32,42,44,51,56,57,65,70]. Humans were tested for OROV antibodies (11 studies)[5,18,[38][39][40]58,63,64,68,69,78], OROV nucleic acid (19 of studies)[3,6,8,34,43,[47][48][49][53][54][55]59,61,62,66,72,73,76,77], or both (17 studies)[4,7,[15][16][17]24,32,41,…”

mentioning

confidence: 99%

Observational Characterization of the Ecological and Environmental Features Associated with the Presence of Oropouche Virus and the Primary Vector Culicoides paraensis: Data Synthesis and Systematic Review

2021

View full text Add to dashboard Cite

Oropouche virus (OROV), a member of the Orthobunyavirus genus, is an arthropod-borne virus (arbovirus) and is the etiologic agent of human and animal disease. The primary vector of OROV is presumed to be the biting midge, Culicoides paraenesis, though Culex quinquefasciatus, Cq. venezuelensis, and Aedes serratus mosquitoes are considered secondary vectors. The objective of this systematic review is to characterize locations where OROV and/or its primary vector have been detected. Synthesis of known data through review of published literature regarding OROV and vectors was carried out through two independent searches: one search targeted to OROV, and another targeted towards the primary vector. A total of 911 records were returned, but only 90 (9.9%) articles satisfied all inclusion criteria. When locations were characterized, some common features were noted more frequently than others, though no one characteristic was significantly associated with presence of OROV using a logistic classification model. In a separate correlation analysis, vector presence was significantly correlated only with the presence of restingas. The lack of significant relationships is likely due to the paucity of data regarding OROV and its eco-epidemiology and highlights the importance of continued focus on characterizing this and other neglected tropical diseases.

show abstract