Determination of Viremia and Concentration of Circulating Nonstructural Protein 1 in Patients Infected with Dengue Virus in Mexico

Cruz-Hernández, Sergio I. de la; Flores-Aguilar, Hilario; González-Mateos, Silvia; López-Martı́nez, Irma; Alpuche-Aranda, Celia; Ludert, Juan E.; Ángel, Rosa M. del

doi:10.4269/ajtmh.12-0023

Cited by 74 publications

(70 citation statements)

References 36 publications

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“…A previous study also observed that there was no evidence of positive selection in the structural proteins of DENV-2 including the NS1 protein [29]. This indicates that NS1 needs to retain its structure and amino acid sequences to be able to perform its function effectively.…”

Section: Discussionmentioning

confidence: 91%

Performance of commercial dengue NS1 ELISA and molecular analysis of NS1 gene of dengue viruses obtained during surveillance in Indonesia

et al. 2013

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BackgroundEarly diagnosis of dengue infection is crucial for better management of the disease. Diagnostic tests based on the detection of dengue virus (DENV) Non Structural Protein 1 (NS1) antigen are commercially available with different sensitivities and specificities observed in various settings. Dengue is endemic in Indonesia and clinicians are increasingly using the NS1 detection for dengue confirmation. This study described the performance of Panbio Dengue Early NS1 and IgM Capture ELISA assays for dengue detection during our surveillance in eight cities in Indonesia as well as the genetic diversity of DENV NS1 genes and its relationship with the NS1 detection.MethodsThe NS1 and IgM/IgG ELISA assays were used for screening and confirmation of dengue infection during surveillance in 2010–2012. Collected serum samples (n = 440) were subjected to RT-PCR and virus isolation, in which 188 samples were confirmed for dengue infection. The positivity of the ELISA assays were correlated with the RT-PCR results to obtain the sensitivity of the assays. The NS1 genes of 48 Indonesian virus isolates were sequenced and their genetic characteristics were studied.ResultsUsing molecular data as gold standard, the sensitivity of NS1 ELISA assay for samples from Indonesia was 56.4% while IgM ELISA was 73.7%. When both NS1 and IgM results were combined, the sensitivity increased to 89.4%. The NS1 sensitivity varied when correlated with city/geographical origins and DENV serotype, in which the lowest sensitivity was observed for DENV-4 (19.0%). NS1 sensitivity was higher in primary (67.6%) compared to secondary infection (48.2%). The specificity of NS1 assay for non-dengue samples were 100%. The NS1 gene sequence analysis of 48 isolates revealed the presence of polymorphisms of the NS1 genes which apparently did not influence the NS1 sensitivity.ConclusionsWe observed a relatively low sensitivity of NS1 ELISA for dengue detection on RT-PCR-positive dengue samples. The detection rate increased significantly when NS1 data was combined with IgM. In our study, the low sensitivity of NS1 antigen detection did not relate to NS1 genetic diversity. Rather, the performance of the NS1 antigen test was affected by the infection status of patients and geographical origin of samples.

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Section: Discussionmentioning

confidence: 91%

Performance of commercial dengue NS1 ELISA and molecular analysis of NS1 gene of dengue viruses obtained during surveillance in Indonesia

et al. 2013

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“…We first validated our assay and demonstrated dose-response curves in buffer ( Figure 3) for each dengue serotype. We could demonstrate a lower limit of detection (LoD) of 0.5 ng/mL (DENV-1), 0.2 ng/mL (DENV-2), 0.8 ng/mL (DENV-3) and 1 ng/mL (DENV-4), well below reported levels of circulating NS1 in patients affected from primary or secondary infection (reference values of circulating NS1: 13 ng/mL -1880 ng/mL for patients infected with DENV-1 and 4 ng/mL -266 ng/mL for patients infected with DENV-2 in a cohort study of 225 patients) 37 . We were able to demonstrate sensitive and selective detection of each dengue serotype with minimal cross talk between serotypes.…”

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confidence: 85%

Improved Immunoassay Sensitivity in Serum as a Result of Polymer-Entrapped Quantum Dots: ‘Papaya Particles’

et al. 2015

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Fluorescent labels are widely employed in biomarker quantification and diagnostics, however they possess narrow Stokes shifts and can photobleach, limiting multiplexed detection applications and compromising sensitivity. In contrast, quantum dots do not photobleach and have much wider Stokes shifts, but a paucity of robust surface attachment chemistries for bioconjugation has limited their uptake in biomedical diagnostics. We report a novel class of biofunctional fluorescent labels based on trapping of ∼10(4) quantum dots within a core nanoparticle. The doped particles act as scaffolds for generation of a multilayered shell consisting of a functionalized hydrophilic polymer with covalently attached receptors for analyte capture. These constructs, which conceptually resemble a papaya fruit, are chemically stable, remain monodispersed for >6 months in buffer, and show utility in immunoassay applications. Using monoclonal antibody fragments against nonstructural protein dengue NS1, an early biomarker for dengue fever, antibody immobilization capacity was 75-fold higher compared with traditional carbodiimide protein coupling. In the model dengue immunoassay, we observed a 15-fold lower limit of detection and 4-fold higher fluorescence intensity with the "papaya particles" compared to current "best-in-class" commercial reagents. Direct deployment in human serum allowed sensitive detection of different NS1 serotypes with lower limits of detection within the clinically relevant range (1-10 ng/mL), and sufficient specificity for identification of the dengue serotype was achieved for concentrations >10 ng/mL (DV1-3) and >50 ng/mL (DV4). The combination of chemical and physical stability and high binding capacity combined with the intrinsic advantages of quantum dots may enable more simple, robust diagnostic assays in the future.

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“…In some cases, it has been described that there is no association between viremia and NS1 levels in severe clinical condition [78,79]. In addition, the infected serotype might have a role in diagnosis sensitivity; it has been reported that NS1 levels are higher in patients infected with DENV1 than with DENV2 [76,78].…”

Section: Differential Diagnosismentioning

confidence: 98%

“…Although it has been described that the highest levels of viremia and NS1 are observed in the first 2-3 days after fever onset, it is well defined that both parameters are higher in patients with secondary DENV infections and the severe form of dengue [39,[73][74][75][76][77]. In some cases, it has been described that there is no association between viremia and NS1 levels in severe clinical condition [78,79]. In addition, the infected serotype might have a role in diagnosis sensitivity; it has been reported that NS1 levels are higher in patients infected with DENV1 than with DENV2 [76,78].…”

Section: Differential Diagnosismentioning

confidence: 98%

“…Although viremia has been associated with the severity of dengue clinical syndrome [74,82], its determination is a laborintensive task and alternatives are needed. In a number of cases, a correlation between viremia and NS1 antigenemia levels have been found and the association of both parameters with dengue prognosis and disease progression has been described [39,74,78].…”

Section: Future Perspectivementioning

confidence: 99%

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Dengue Laboratory Diagnosis: Still Some Room for Improvement

et al. 2015

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Dengue is the most important and widely distributed arthropod-borne viral disease affecting humans. The number of dengue virus infections has steadily grown and more than 100 countries survey dengue incidence every year. Due to the lack of an approved antiviral treatment or licensed preventative vaccine, accurate and opportune diagnosis is commended for efficient dengue epidemiological surveillance, to propose control measures in order to curtail outbreaks timely and treat patients satisfactorily. In this review, the basis, application and indications for different diagnostic tests are described, and their advantages and limitations considered. At the end of this piece, we speculate what the future may hold for the diagnosis of dengue infections. KeywordsThe four serotypes of dengue virus (DENV 1-4) may equally be the cause of dengue infection in humans. DENV is a member of the Flaviviridae family, genus Flavivirus and is transmitted by the bite of female mosquitoes of the Aedes genus, mainly Aedes aegypti; an anthropophilic arthropod extensively distributed [1,2]. The viral genome is constituted by a single RNA strand of positive polarity of approximately 11 kb. It encodes three structural proteins: the envelope (E), pre-membrane/membrane (prM/M) and capsid (C) proteins; and seven nonstructural (NS) proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B and NS5), which are implicated in viral translation, replication and modulation of the host immune response [2,3]. Dengue prevails in 128 countries and it is estimated that more than 2.5 billion people (two fifths of the world's population) are at risk of DENV infection [1,2]. Every year over 50 million DENV infections occur, causing more than 500,000 severe cases of dengue and resulting in at least 125,000 fatalities. The enormous increase in dengue prevalence observed in the last decades is multifactorial in nature. Unplanned and uncontrolled urbanization caused by the global population growth is associated with the deterioration in water supply quality and poor sewer and waste management systems availability. These conditions are ideal for the domestic propagation of the vector and a hallmark for the most recent epidemic outbreaks observed in tropical regions of the world such as Southeast Asia and South America. In addition, the modern trade globalization facilitates the introduction of the vector and all DENV serotypes to previously uninfected populated centers [1,4,5], where a highly dense susceptible population may be at risk. In this way, over the last decades DENV infection has become a global menace not only for tropical regions of the world, where the vector thrives, but also for temperate zones starting to be colonized by Aedes mosquitoes. The annual cost of dengue disease is estimated at US$135 million in Asiatic countries alone [6].

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Determination of Viremia and Concentration of Circulating Nonstructural Protein 1 in Patients Infected with Dengue Virus in Mexico

Cited by 74 publications

References 36 publications

Performance of commercial dengue NS1 ELISA and molecular analysis of NS1 gene of dengue viruses obtained during surveillance in Indonesia

Performance of commercial dengue NS1 ELISA and molecular analysis of NS1 gene of dengue viruses obtained during surveillance in Indonesia

Improved Immunoassay Sensitivity in Serum as a Result of Polymer-Entrapped Quantum Dots: ‘Papaya Particles’

Dengue Laboratory Diagnosis: Still Some Room for Improvement

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