Improving dengue viral antigens detection in dengue patient serum specimens using a low pH glycine buffer treatment

Shen, Wen-Fan; Galula, Jedhan Ucat; Chang, Gwong‐Jen J.; Wu, Han‐Chung; King, Chwan-Chuen; Chao, Day‐Yu

doi:10.1016/j.jmii.2015.05.008

Cited by 14 publications

(6 citation statements)

References 29 publications

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“…In this context, a number of developed immunosensors have shown excellent sensitivity. For example, Dias and co-workers reported 0.12 ng/mL while Cavalcanti and co-workers reported 0.33ng/mL limit of detection for their immunosensors, which is lower than 7ng/mL and 3.04 ng/mL limit of detection reported in some ELISA-based assays [ 73 , 74 ].…”

Section: Laboratory Diagnosis and Its Significancementioning

confidence: 87%

Diagnosis of Dengue Infection Using Conventional and Biosensor Based Techniques

Parkash

Shueb

2015

Viruses

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Dengue is an arthropod-borne viral disease caused by four antigenically different serotypes of dengue virus. This disease is considered as a major public health concern around the world. Currently, there is no licensed vaccine or antiviral drug available for the prevention and treatment of dengue disease. Moreover, clinical features of dengue are indistinguishable from other infectious diseases such as malaria, chikungunya, rickettsia and leptospira. Therefore, prompt and accurate laboratory diagnostic test is urgently required for disease confirmation and patient triage. The traditional diagnostic techniques for the dengue virus are viral detection in cell culture, serological testing, and RNA amplification using reverse transcriptase PCR. This paper discusses the conventional laboratory methods used for the diagnosis of dengue during the acute and convalescent phase and highlights the advantages and limitations of these routine laboratory tests. Subsequently, the biosensor based assays developed using various transducers for the detection of dengue are also reviewed.

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Section: Laboratory Diagnosis and Its Significancementioning

confidence: 87%

Diagnosis of Dengue Infection Using Conventional and Biosensor Based Techniques

Parkash

Shueb

2015

Viruses

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“…These complexes may hinder epitope recognition by anti-NS1 antibodies in the NS1 assay. Dissociation of these immune complexes by various pretreatment methods (for example, heat or acid treatment) has been shown to enhance the sensitivity of DENV NS1 assays [49,[51][52][53]. However, to our experiences, we observed that heat-or acid-pretreated specimens sometimes altered serotype-specific interpretation to be negative or cross reactive to other serotypes, possibly due to the disruption of epitopes recognized by conformation-dependent antibodies.…”

Section: Plos Neglected Tropical Diseasesmentioning

confidence: 47%

High performance dengue virus antigen-based serotyping-NS1-ELISA (plus): A simple alternative approach to identify dengue virus serotypes in acute dengue specimens

et al. 2021

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Dengue hemorrhagic fever (DHF) is caused by infection with dengue virus (DENV). Four different serotypes (DENV1-4) co-circulate in dengue endemic areas. The viral RNA genome-based reverse-transcription PCR (RT-PCR) is the most widely used method to identify DENV serotypes in patient specimens. However, the non-structural protein 1 (NS1) antigen as a biomarker for DENV serotyping is an emerging alternative method. We modified the serotyping-NS1-enzyme linked immunosorbent assay (stNS1-ELISA) from the originally established assay which had limited sensitivity overall and poor specificity for the DENV2 serotype. Here, four biotinylated serotype-specific antibodies were applied, including an entirely new design for detection of DENV2. Prediction of the infecting serotype of retrospective acute-phase plasma from dengue patients revealed 100% concordance with the standard RT-PCR method for all four serotypes and 78% overall sensitivity (156/200). The sensitivity of DENV1 NS1 detection was greatly improved (from 62% to 90%) by the addition of a DENV1/DENV3 sub-complex antibody pair. Inclusive of five antibody pairs, the stNS1-ELISA (plus) method showed an overall increased sensitivity to 85.5% (171/200). With the same clinical specimens, a commercial NS1 rapid diagnostic test (NS1-RDT) showed 72% sensitivity (147/200), significantly lower than the stNS1-ELISA (plus) performance. In conclusion, the stNS1-ELISA (plus) is an improved method for prediction of DENV serotype and for overall sensitivity. It could be an alternative assay not only for early dengue diagnosis, but also for serotype identification especially in remote resource-limited dengue endemic areas.

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“…This is potentially due to the presence of NS1 immune complex (IC) which is generated by circulating NS1 and existing anti-NS1 antibodies [ 50 , 51 , 52 ] which reduces the amount of free NS1 protein. Disassociation of NS1 IC by heat or acid/base methods explored in previous studies [ 53 , 54 , 55 ] were found to enhance assay sensitivity. However, our NS1-Cygnus device and NS1-microplate ELISA gave above 82% sensitivity despite patient plasma used in this study being mostly from secondary infections (98%).…”

Section: Discussionmentioning

confidence: 94%

Smartphone multiplex microcapillary diagnostics using Cygnus: Development and evaluation of rapid serotype-specific NS1 detection with dengue patient samples

et al. 2022

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Laboratory diagnosis of dengue virus (DENV) infection including DENV serotyping requires skilled labor and well-equipped settings. DENV NS1 lateral flow rapid test (LFT) provides simplicity but lacks ability to identify serotype. A simple, economical, point-of-care device for serotyping is still needed. We present a gravity driven, smartphone compatible, microfluidic device using microcapillary film (MCF) to perform multiplex serotype-specific immunoassay detection of dengue virus NS1. A novel device–termed Cygnus–with a stackable design allows analysis of 1 to 12 samples in parallel in 40 minutes. A sandwich enzyme immunoassay was developed to specifically detect NS1 of all four DENV serotypes in one 60-μl plasma sample. This test aims to bridge the gap between rapid LFT and laboratory microplate ELISAs in terms of sensitivity, usability, accessibility and speed. The Cygnus NS1 assay was evaluated with retrospective undiluted plasma samples from 205 DENV infected patients alongside 50 febrile illness negative controls. Against the gold standard RT-PCR, clinical sensitivity for Cygnus was 82% in overall (with 78, 78, 80 and 76% for DENV1-4, respectively), comparable to an in-house serotyping NS1 microplate ELISA (82% vs 83%) but superior to commercial NS1-LFT (82% vs 74%). Specificity of the Cygnus device was 86%, lower than that of NS1-microplate ELISA and NS1-LFT (100% and 98%, respectively). For Cygnus positive samples, identification of DENV serotypes DENV2-4 matched those by RT-PCR by 100%, but for DENV1 capillaries false positives were seen, suggesting an improved DENV1 capture antibody is needed to increase specificity. Overall performance of Cygnus showed substantial agreement to NS1-microplate ELISA (κ = 0.68, 95%CI 0.58–0.77) and NS1-LFT (κ = 0.71, 95%CI 0.63–0.80). Although further refinement for DENV-1 NS1 detection is needed, the advantages of multiplexing and rapid processing time, this Cygnus device could deliver point-of-care NS1 antigen testing including serotyping for timely DENV diagnosis for epidemic surveillance and outbreak prediction.

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Improving dengue viral antigens detection in dengue patient serum specimens using a low pH glycine buffer treatment

Abstract: Inclusion of a low-pH glycine buffer treatment step in the commercially available Ag-ELISA is crucial for clinical diagnosis and E-containing viral particles could be a valuable target for acute DENV diagnosis, similar to NS1 detection.

Cited by 14 publications

References 29 publications

Diagnosis of Dengue Infection Using Conventional and Biosensor Based Techniques

Diagnosis of Dengue Infection Using Conventional and Biosensor Based Techniques

High performance dengue virus antigen-based serotyping-NS1-ELISA (plus): A simple alternative approach to identify dengue virus serotypes in acute dengue specimens

Smartphone multiplex microcapillary diagnostics using Cygnus: Development and evaluation of rapid serotype-specific NS1 detection with dengue patient samples

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