Development of multiplex real‐time reverse‐transcriptase polymerase chain reaction assay for simultaneous detection of Zika, dengue, yellow fever, and chikungunya viruses in a single tube

Wu, Wenhui; Wang, Jin; Yu, Nan; Yan, Juying; Zhuo, Zhihao; Su, Xiaosong; Fang, Mujin; He, Shuizhen; Zhang, Yanjun; Ge, Shuping; Xia, Ningshao

doi:10.1002/jmv.25253

Cited by 33 publications

(39 citation statements)

References 26 publications

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“…The EV71, an unenveloped RNA virus, was included as negative control. The primers used to detect the RNA sequences of ZIKV, DENV-2, YFV, and EV71 were described previously (Lok et al, 2012;Fernandes-Monteiro et al, 2015;Fischer et al, 2017;Yu et al, 2017;Wu et al, 2018;Yuan et al, 2018) and listed in Table 1.…”

Section: Rnase Digestion Assay and Quantitative Reverse Transcriptionmentioning

confidence: 99%

Montelukast, an Anti-asthmatic Drug, Inhibits Zika Virus Infection by Disrupting Viral Integrity

Chen

Wang

et al. 2020

Front. Microbiol.

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The association of Zika virus (ZIKV) infection and severe complications including neurological sequelae especially fetal microcephaly has aroused global attentions since its outbreak in 2015. Currently, there are no vaccines or therapeutic drugs clinically approved for treatments of ZIKV infection, however. And the drugs used for treating ZIKV in pregnant women require a higher safety profile. Here, we identified an anti-asthmatic drug, montelukast, which is of safety profile for pregnant women and exhibited antiviral efficacy against ZIKV infection in vitro and in vivo. And we showed that montelukast could disrupt the integrity of the virions to release the viral genomic RNA, hence irreversibly inhibiting viral infectivity. In consideration of the neuro-protective activity that montelukast possessed, which was previously reported, it is promising that montelukast could be used for patients with ZIKV infection, particularly for pregnant women.

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Section: Rnase Digestion Assay and Quantitative Reverse Transcriptionmentioning

confidence: 99%

Montelukast, an Anti-asthmatic Drug, Inhibits Zika Virus Infection by Disrupting Viral Integrity

Chen

Wang

et al. 2020

Front. Microbiol.

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“…A variety of molecular assays for CHIKV have been published or are commercially available (see Table S1 in the supplemental material). Reported assays include conventional RT-PCR (69, 108-112), real-time RT-PCR (rRT-PCR) (102,105,(113)(114)(115)(116)(117)(118)(119)(120), isothermal methods (110,(121)(122)(123)(124), and multiplex assays (125)(126)(127)(128)(129)(130)(131)(132)(133)(134)(135)(136)(137)(138). No molecular gold standard by which to evaluate reported assays in practice exists, and the decision to implement a particular test depends on the relative advantages and disadvantages of the method along with the capabilities in a given laboratory.…”

Section: Chikv Diagnosticsmentioning

confidence: 99%

Beyond Fever and Pain: Diagnostic Methods for Chikungunya Virus

2019

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Chikungunya virus (CHIKV) is an alphavirus that is primarily transmitted by Aedes species mosquitoes. Though reports of an illness consistent with chikungunya date back over 200 years, CHIKV only gained worldwide attention during a massive pandemic that began in East Africa in 2004. Chikungunya, the clinical illness caused by CHIKV, is characterized by a rapid onset of high fever and debilitating joint pain, though in practice, etiologic confirmation of CHIKV requires the availability and use of specific laboratory diagnostics. Similar to infections caused by other arboviruses, CHIKV infections are most commonly detected with a combination of molecular and serological methods, though cell culture and antigen detection are reported. This review provides an overview of available CHIKV diagnostics and highlights aspects of basic virology and epidemiology that pertain to viral detection. Although the number of chikungunya cases has decreased since 2014, CHIKV has become endemic in countries across the tropics and will continue to cause sporadic outbreaks in naive individuals. Consistent access to accurate diagnostics is needed to detect individual cases and initiate timely responses to new outbreaks.

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“…The specificity of the ZIKV and DENV2 assays was then determined using a cross-reactivity panel. To address challenges involving primer cross-reactivity with other arboviruses (46, 48) as well as the tendency toward false-positive amplification in negative samples, we tested the specificity of the assay against a panel of Flaviviruses and one Alphavirus . The frequently co-circulating arboviral RNA samples, obtained from BEI resources (Table 2) for this study, were: CHIKV (H20235 ST MARTIN 2013), JEV (India R53567), WNV (CO 1862), and YFV (17D).…”

Section: Resultsmentioning

confidence: 99%

“…We then employed a crude extraction method using squash buffer to extract viral DENV2 and ZIKV RNA from mosquitoes for use in the bCUBE system. To address challenges involving primer cross-reactivity with other arboviruses (46, 48) as well as the tendency toward false-positive amplification in negative samples, we tested the specificity of the assay against a panel of frequently co-circulating arboviruses. The results confirmed the assay’s specificity for DENV2 and ZIKV RNA, with the potential to apply these primer pairs for field conditions.…”

Section: Discussionmentioning

confidence: 99%

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Field-deployable molecular diagnostic platform for arbovirus andWolbachiadetection inAedes aegypti

Rutkowski

Dong

Dimopoulos

2020

Preprint

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26Background: Surveillance of mosquito infection status is critical for planning and deployment 27 of proper mosquito control initiatives. Concurrently, Wolbachia is being widely used as a control 28 method for arboviral transmission. Point-of-care (POC) detection assays are necessary for 29 monitoring the infection prevalence and geographic range of viruses as well as Wolbachia in 30 mosquito vector populations. We therefore assessed the novel qPCR bCUBE molecular 31 diagnostic system as a tool for virus and Wolbachia detection. 32 Results: We developed a reliable, specific, and sensitive diagnostic assay for detecting Zika 33 virus and dengue virus serotype 2 using the real-time qPCR platform bCUBE. With bCUBE-34 based qRT-PCR, both Wolbachia bacterium and virus RNA could be reliably detected in 35 individually infected Ae. aegypti mosquitoes and in pools of 5, 10, or 15 mosquitoes. 36 Conclusions: The portable qPCR bCUBE diagnostic platform is capable of detecting Zika and 37 dengue virus as well as Wolbachia in mosquitoes and therefore has potential as a practical field-38 deployable diagnostic test for vector-borne disease surveillance programs. 39 40 41 42 43 44 45 46 47

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Development of multiplex real‐time reverse‐transcriptase polymerase chain reaction assay for simultaneous detection of Zika, dengue, yellow fever, and chikungunya viruses in a single tube

Cited by 33 publications

References 26 publications

Montelukast, an Anti-asthmatic Drug, Inhibits Zika Virus Infection by Disrupting Viral Integrity

Montelukast, an Anti-asthmatic Drug, Inhibits Zika Virus Infection by Disrupting Viral Integrity

Beyond Fever and Pain: Diagnostic Methods for Chikungunya Virus

Field-deployable molecular diagnostic platform for arbovirus andWolbachiadetection inAedes aegypti

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