A Dry-format Field-deployable Quantitative Reverse Transcriptase-polymerase Chain Reaction Assay for Diagnosis of Dengue Infections

Wu, Shuenn-Jue; Pal, Subhamoy; Ekanayake, Sajeewane; Greenwald, David A.; Lara, Silvia Hunold; Raviprakash, Kanakatte; Kochel, Tadeusz J.; Porter, Kevin R.; Hayes, Curtis G.; Nelson, William M.; Callahan, Johnny D.

doi:10.4269/ajtmh.2008.79.505

Cited by 14 publications

(11 citation statements)

References 14 publications

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“…The developed rRT-PCR assay allowed the detection of 0.5 pfu/ml. This detection limit value is similar to that found by Wu et al, [29] and is therefore sensitive enough to diagnose ZIKV in clinical cases. Indeed, viraemia found in human infection ranges from 10 2 to 10 6 pfu/ml [8,10,21,32].…”

Section: Discussionsupporting

confidence: 88%

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Quantitative real-time PCR detection of Zika virus and evaluation with field-caught Mosquitoes

Faye

Diallo

et al. 2013

Virol J

346

270

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BackgroundZika virus (ZIKV), a mosquito borne flavivirus is a pathogen affecting humans in Asia and Africa. ZIKV infection diagnosis relies on serology–which is challenging due to cross-reactions with other flaviviruses and/or absence or low titer of IgM and IgG antibodies at early phase of infection- virus isolation, which is labor intensive, time consuming and requires appropriate containment. Therefore, real-time RT-PCR (rRT-PCR) is an appealing option as a rapid, sensitive and specific method for detection of ZIKV in the early stage of infection. So far, only one rRT-PCR assay has been described in the context of the outbreak in Micronesia in 2007. In this study, we described a one step rRT-PCR for ZIKV which can detect a wider genetic diversity of ZIKV isolates from Asia and Africa.ResultsThe NS5 protein coding regions of African ZIKV isolates were sequenced and aligned with representative flaviviruses sequences from GenBank to design primers and probe from conserved regions. The analytical sensitivity of the assay was evaluated to be 32 genome-equivalents and 0.05 plaque forming unit (pfu). The assay was shown to detect 37 ZIKV isolates covering a wide geographic in Africa and Asia over 36 years but none of the 31 other flaviviruses tested showing high analytical specificity. The rRT-PCR could be performed in less than 3 hours. This method was used successfully to detect ZIKV strains from field-caught mosquitoes.ConclusionWe have developed a rapid, sensitive and specific rRT – PCR for detection of ZIKV. This assay is a useful tool for detection of ZIKV infection in regions where a number of other clinically indistinguishable arboviruses like dengue or chikungunya co-circulate. Further studies are needed to validate this assay in clinical positive samples collected during acute ZIKV infection.

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Section: Discussionsupporting

confidence: 88%

“…The mosquitoes pools were also screened for dengue an yellow fever using primers and probes described previously [28,29]. …”

Section: Methodsmentioning

confidence: 99%

Quantitative real-time PCR detection of Zika virus and evaluation with field-caught Mosquitoes

Faye

Diallo

et al. 2013

Virol J

346

270

View full text Add to dashboard Cite

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“…The standard curve was completed by serially diluting the virus stock and extracting the RNA from each dilution according to the previously mentioned RNA extraction protocol while simultaneously titrating each dilution in a standard plaque assay (PFU/ml). A curve correlation coefficient of Ն0.950 and a 90 to 100% PCR efficiency was used to validate each detection assay, and the RNA amounts were correlated with numbers of PFU equivalents per milliliter as previously reported (22,45). While an alternative approach is to calculate numbers of RNA copies per ml, we chose the presentation of PFU equivalents per ml as this is more relevant in a diagnostic setting.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of Widely Used Diagnostic Tests To Detect West Nile Virus Infections in Horses Previously Infected with St. Louis Encephalitis Virus or Dengue Virus Type 2

Ledermann

Loroño-Pino

Ellis

et al. 2011

Clin Vaccine Immunol

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Primary West Nile virus (WNV) infections can be diagnosed using a number of tests that detect infectious particles, nucleic acid, and specific IgM and/or IgG antibodies. However, serological identification of the infecting agent in secondary or subsequent flavivirus infections is problematic due to the extensive cross-reactivity of flavivirus antibodies. This is particularly difficult in the tropical Americas where multiple flaviviruses cocirculate. A study of sequential flavivirus infection in horses was undertaken using three medically important flaviviruses and five widely utilized diagnostic assays to determine if WNV infection in horses that had a previous St. Louis encephalitis virus (SLEV) or dengue virus type 2 (DENV-2) infection could be diagnosed. Following the primary inoculation, 25% (3/12) and 75% (3/4) of the horses mounted antibody responses against SLEV and DENV-2, respectively. Eighty-eight percent of horses subsequently inoculated with WNV had a WNV-specific antibody response that could be detected with one of these assays. The plaque reduction neutralization test (PRNT) was sensitive in detection but lacked specificity, especially following repeated flavivirus exposure. The WNV-specific IgM enzyme-linked immunosorbent assay (IgM ELISA) was able to detect an IgM antibody response and was not crossreactive in a primary SLEV or DENV response. The WNV-specific blocking ELISA was specific, showing positives only following a WNV injection. Of great importance, we demonstrated that timing of sample collection and the need for multiple samples are important, as the infecting etiology could be misdiagnosed if only a single sample is tested.

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“…In addition to weather, the impact of the dynamics of circulation of dengue virus serotypes on dengue epidemiology has been well documented [28]. Infection with one serotype confers life-long immunity to that particular serotype [29], [30]. Some studies have also reported a time-lagged correlation between dengue virus serotype dynamics and disease incidence rates [31].…”

Section: Introductionmentioning

confidence: 99%

Statistical Modeling Reveals the Effect of Absolute Humidity on Dengue in Singapore

Lee

et al. 2014

PLoS Negl Trop Dis

100

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Weather factors are widely studied for their effects on indicating dengue incidence trends. However, these studies have been limited due to the complex epidemiology of dengue, which involves dynamic interplay of multiple factors such as herd immunity within a population, distinct serotypes of the virus, environmental factors and intervention programs. In this study, we investigate the impact of weather factors on dengue in Singapore, considering the disease epidemiology and profile of virus serotypes. A Poisson regression combined with Distributed Lag Non-linear Model (DLNM) was used to evaluate and compare the impact of weekly Absolute Humidity (AH) and other weather factors (mean temperature, minimum temperature, maximum temperature, rainfall, relative humidity and wind speed) on dengue incidence from 2001 to 2009. The same analysis was also performed on three sub-periods, defined by predominant circulating serotypes. The performance of DLNM regression models were then evaluated through the Akaike's Information Criterion. From the correlation and DLNM regression modeling analyses of the studied period, AH was found to be a better predictor for modeling dengue incidence than the other unique weather variables. Whilst mean temperature (MeanT) also showed significant correlation with dengue incidence, the relationship between AH or MeanT and dengue incidence, however, varied in the three sub-periods. Our results showed that AH had a more stable impact on dengue incidence than temperature when virological factors were taken into consideration. AH appeared to be the most consistent factor in modeling dengue incidence in Singapore. Considering the changes in dominant serotypes, the improvements in vector control programs and the inconsistent weather patterns observed in the sub-periods, the impact of weather on dengue is modulated by these other factors. Future studies on the impact of climate change on dengue need to take all the other contributing factors into consideration in order to make meaningful public policy recommendations.

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A Dry-format Field-deployable Quantitative Reverse Transcriptase-polymerase Chain Reaction Assay for Diagnosis of Dengue Infections

Cited by 14 publications

References 14 publications

Quantitative real-time PCR detection of Zika virus and evaluation with field-caught Mosquitoes

Quantitative real-time PCR detection of Zika virus and evaluation with field-caught Mosquitoes

Evaluation of Widely Used Diagnostic Tests To Detect West Nile Virus Infections in Horses Previously Infected with St. Louis Encephalitis Virus or Dengue Virus Type 2

Statistical Modeling Reveals the Effect of Absolute Humidity on Dengue in Singapore

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