Simultaneous detection and quantitation of Chikungunya, Dengue and West Nile viruses by multiplex RT-PCR assays and Dengue virus typing using High Resolution Melting

Naze, Florence; Roux, K. Le; Schuffenecker, Isabelle; Zeller, H.; Staïkowsky, F.; Grivard, Philippe; Michault, Alain; Laurent, Philippe

doi:10.1016/j.jviromet.2009.03.006

Cited by 58 publications

(47 citation statements)

References 30 publications

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“…The IC RNA included in the assay is amplified competitively with the same set of primers dedicated to the specific amplification of the EV template. Another major interest of the IC RNA is its incorporation directly in the biological specimens to be tested before RNA extraction, which allows close monitoring of the entire sample process (Naze et al, 2009;Villanova et al, 2007;Xiao et al, 2009). Because of its competitive property, particular attention has been paid to the molecular design of IC to avoid cross hybridization with the EV template genome and to ensure optimal amplification of both IC and EV templates.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Quantitative real-time RT-PCR assay for research studies on enterovirus infections in the central nervous system

Volle

Nourrisson

Mirand

et al. 2012

Journal of Virological Methods

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Section: Discussionmentioning

confidence: 99%

“…Internal control (IC) is an RNA chimera synthesized using a previously described method with minor modifications (Naze et al, 2009). Briefly, EV1pBR and EV2pBR primers were used to amplify a 217-bp segment chosen in the plasmid vector pBR322.…”

Section: Internal Control Constructionmentioning

confidence: 99%

Quantitative real-time RT-PCR assay for research studies on enterovirus infections in the central nervous system

Volle

Nourrisson

Mirand

et al. 2012

Journal of Virological Methods

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“…Though numerous DENV species-specific and serotype-specific assays have since been developed, direct comparisons between these assays are notably rare (11,(13)(14)(15)(16)(17). Rather, studies evaluating new DENV molecular diagnostic tests typically use samples collected within the first 5 days of fever from patients who had positive testing by viral isolation, seroconversion, or both (10,(18)(19)(20)(21)(22)(23)(24)(25). This practice ensures that only the highest viral loads are evaluated and it likely does not reflect clinical reality, where patients may present at any time during their illness, including five or more days after fever onset.…”

mentioning

confidence: 99%

“…Despite the large number of reported DENV NAATs, few of these assays have been designed to include an internal control (IC), either as an extrinsic molecule spiked into each sample before or after extraction or as a heterologous intrinsic target that is coextracted with DENV RNA (14,17,21,25,31). It has been advocated that ICs be used in settings where PCR inhibitors present a significant source of false-negative results, which may be particularly important in the performance of NAATs in countries where dengue is endemic (26,32).…”

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confidence: 99%

Development of an Internally Controlled Real-Time Reverse Transcriptase PCR Assay for Pan-Dengue Virus Detection and Comparison of Four Molecular Dengue Virus Detection Assays

et al. 2013

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A number of diagnostic tests are available for dengue virus (DENV) detection, including a variety of nucleic acid amplification tests (NAATs). However, reports describing a direct comparison of different NAATs have been limited. In this study, we report the design of an internally controlled real-time reverse transcriptase PCR (rRT-PCR) that detects all four DENV serotypes but does not distinguish between them (the pan-DENV assay). Two hundred clinical samples were then tested using four different DENV RT-PCR assays: the pan-DENV assay, a commercially produced, internally controlled DENV rRT-PCR (the Altona assay), a widely used heminested RT-PCR, and a serotype-specific multiplex rRT-PCR assay. The pan-DENV assay had a linear range extending from 1.0 to 7.0 log 10 cDNA equivalents/l and a lower limit of 95% detection ranging from 1.7 to 7.6 cDNA equivalents/l, depending on the serotype. When measured against a composite reference standard, the pan-DENV assay proved to be more clinically sensitive than either the Altona or heminested assays, with a sensitivity of 98.0% compared to 72.3% and 78.8%, respectively (P < 0.0001 for both comparisons). The pan-DENV assay detected DENV in significantly more samples collected on or after day 5 of illness and in a subgroup of patients with detectable anti-DENV IgM at presentation. No significant difference in sensitivity was observed between the pan-DENV assay and the multiplex rRT-PCR, despite the presence of an internal control in the former. The detection of DENV RNA late in the course of clinical illness should serve to lengthen the period during which a confirmed molecular diagnosis of DENV infection can be provided. Dengue virus (DENV) is the most common vector-borne human viral pathogen worldwide (1). Infection with one or more of the four closely related virus serotypes (designated DENV-1 to -4) results in a range of clinical manifestations spanning asymptomatic infection, dengue fever (DF), and severe dengue, a category that includes entities previously classified as dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) (1, 2). Infection with one serotype (primary infection) results in immunity to that serotype, but infection can still occur with any of the remaining serotypes (secondary infection). Secondary DENV infection has been shown to be a significant risk factor for the development of DHF or DSS (3, 4). DENV transmission largely occurs in the tropical and subtropical regions of the world, though the number of countries where DENV is endemic has been increasing (1, 5). Recent reports estimate that 230 million DENV infections occur annually worldwide, including 2 million cases of severe disease and 21,000 deaths (6). Over 3.6 billion people live in regions where this pathogen is endemic and are at risk for infection (6). DF is also one of the most common causes of a systemic febrile illness in travelers returning from countries where the virus is endemic and remains a major concern for military personnel stationed in these areas (7-9).A wide array o...

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“…Dengue RT-PCR can serotype the four different dengue viruses and can also be multiplexed for simultaneous detection of Chikungunya and West Nile viruses [74]. Dengue may also be isolated by viral culture in cell lines, although this method is slow, labor-intensive and requires specialized laboratory equipment and personnel.…”

Section: Diagnosis Of Viral Arthritidesmentioning

confidence: 99%

Viral arthritides

Outhred

Kok

Dwyer

2011

Expert Review of Anti-infective Therapy

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Viral infections may manifest as acute or chronic arthritis. Joint involvement arises from either direct infection of the joint, through an immunological response directed towards the virus or autoimmunity. Epidemiological clues to the diagnosis include geographic location and exposure to vector-borne, blood-borne or sexually transmitted viruses. Although not always possible, it is important to diagnose the pathogenic virus, usually by serology, nucleic acid tests or rarely, viral culture. In general, viral arthritides are self-limiting and treatment is targeted at symptomatic relief. This article focuses on the causes, clinical features, diagnosis and treatment of viral arthritides.

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Simultaneous detection and quantitation of Chikungunya, Dengue and West Nile viruses by multiplex RT-PCR assays and Dengue virus typing using High Resolution Melting

Cited by 58 publications

References 30 publications

Quantitative real-time RT-PCR assay for research studies on enterovirus infections in the central nervous system

Quantitative real-time RT-PCR assay for research studies on enterovirus infections in the central nervous system

Development of an Internally Controlled Real-Time Reverse Transcriptase PCR Assay for Pan-Dengue Virus Detection and Comparison of Four Molecular Dengue Virus Detection Assays

Viral arthritides

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