scholarly journals Utility of a Commercial Nonstructural Protein 1 Antigen Capture Kit as a Dengue Virus Diagnostic Tool

2010 ◽  
Vol 17 (6) ◽  
pp. 949-953 ◽  
Author(s):  
Kovi Bessoff ◽  
Elena Phoutrides ◽  
Mark Delorey ◽  
Luz N. Acosta ◽  
Elizabeth Hunsperger
Keyword(s):  
Dengue Virus ◽  
Acute Phase ◽  
Nonstructural Protein ◽  
Acute Infection ◽  
False Negative ◽  
Diagnostic Tools ◽  
Enzyme Linked Immunosorbent Assay ◽  
Convalescent Phase ◽  
Nonstructural Protein 1 ◽  
Antigen Capture

ABSTRACT Annually, over 2.5 billion people are at risk for infection with dengue virus (DENV), while between 50 and 100 million people contract the infection. There is an urgent need for alternative diagnostic tools that can detect DENV during acute infection. Recent studies have shown that DENV nonstructural protein 1 (NS1) is detectable in the blood as early as the onset of symptoms and persists well into the convalescent phase of the infection. We evaluated the utility of the Bio-Rad Platelia DENV NS1 antigen capture kit in combination with real-time reverse transcriptase PCR (RT-PCR) and an IgM antibody capture enzyme-linked immunosorbent assay (MAC-ELISA) for refining a new algorithm for the diagnosis of acute- or convalescent-phase DENV infection with a single clinical sample. We tested the Bio-Rad kit with three panels of sera. These panels were designed to evaluate the sensitivities of the NS1 kit for (i) early-convalescent-phase samples, (ii) acute-phase samples with false-negative PCR results, and (iii) IgM-negative convalescent-phase samples from patients with confirmed secondary DENV infections. Results show that NS1 can be detected in 22% of serum samples collected more than 10 days after the onset of illness and in 22% of samples that did not elicit an IgM response. Additionally, NS1 was detected in 37% of the tested acute-phase samples with false-negative PCR results, suggesting that NS1 detection may be valuable in increasing the sensitivity of current acute-phase diagnostics. These results will improve diagnosis with a single acute-phase or early-convalescent-phase sample for disease surveillance and clinical diagnosis.

scholarly journals An Antigen Capture Enzyme-Linked Immunosorbent Assay Reveals High Levels of the Dengue Virus Protein NS1 in the Sera of Infected Patients

2000 ◽  
Vol 38 (3) ◽  
pp. 1053-1057 ◽  
Author(s):  
Paul R. Young ◽  
Paige A. Hilditch ◽  
Cheryl Bletchly ◽  
Wendy Halloran
Keyword(s):  
Monoclonal Antibodies ◽  
Dengue Virus ◽  
Acute Phase ◽  
Immunosorbent Assay ◽  
Nonstructural Protein ◽  
Surrogate Marker ◽  
Detection Sensitivity ◽  
Severe Dengue ◽  
Clinical Samples ◽  
Enzyme Linked Immunosorbent Assay

We describe the development of a capture enzyme-linked immunosorbent assay for the detection of the dengue virus nonstructural protein NS1. The assay employs rabbit polyclonal and monoclonal antibodies as the capture and detection antibodies, respectively. Immunoaffinity-purified NS1 derived from dengue 2 virus-infected cells was used as a standard to establish a detection sensitivity of approximately 4 ng/ml for an assay employing monoclonal antibodies recognizing a dengue 2 serotype-specific epitope. A number of serotype cross-reactive monoclonal antibodies were also shown to be suitable probes for the detection of NS1 expressed by the remaining three dengue virus serotypes. Examination of clinical samples demonstrated that the assay was able to detect NS1 with minimal interference from serum components at the test dilutions routinely used, suggesting that it could form the basis of a useful additional diagnostic test for dengue virus infection. Furthermore, quantitation of NS1 levels in patient sera may prove to be a valuable surrogate marker for viremia. Surprisingly high levels of NS1, as much as 15 μg/ml, were found in acute-phase sera taken from some of the patients experiencing serologically confirmed dengue 2 virus secondary infections but was not detected in the convalescent sera of these patients. In contrast, NS1 could not be detected in either acute-phase or convalescent serum samples taken from patients with serologically confirmed primary infection. The presence of high levels of secreted NS1 in the sera of patients experiencing secondary dengue virus infections, and in the context of an anamnestic antibody response, suggests that NS1 may contribute significantly to the formation of the circulating immune complexes that are suspected to play an important role in the pathogenesis of severe dengue disease.

scholarly journals Co-circulation of all the Four Serotype of Dengue Virus in Endemic Region of Saurashtra, Gujarat during 2019-2020 Season

2021 ◽  
Author(s):  
Binita Joseph Aring ◽  
Dipali Magan Bhai Gavali ◽  
Pushpa Ramjibhai Kateshiya ◽  
Hiral Modbhai Gadhvi ◽  
Summaiya Mullan ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Nonstructural Protein ◽  
Geographical Area ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Endemic Region ◽  
Male Population ◽  
Monsoon Period ◽  
Cross Sectional ◽  
Nonstructural Protein 1

Introduction: Dengue has rapidly emerged as a vector - borne viral disease in recent years and also endemic in all continents. The agent of dengue, i.e., dengue viruses, are categorised under the genus Flavivirus, with the four dengue virus serotypes: designated as DENV-1, DENV-2, DENV-3 and DENV-4. These all four serotypes are in circulation either singly, or more than one at the same time. Aim: To study the epidemiological update of dengue with circulating serotype and co-infection in Saurashtra region, Gujarat, India. Materials and Methods: A cross-sectional study was conducted during January 2019 to December 2020 and total samples received were 12,563 which were clinically suspected dengue samples case. After receiving blood samples, serum was separated and proceeded for Dengue NS1Ag (nonstructural protein 1 antigen), and Dengue IgM Ab (Immunoglobulin M antibody). After serological confirmation, 151 samples from different geographical area were selected for Dengue specific Reverse Transcriptase- Polymerase Chain Reaction (RT-PCR) for serotyping. results: Total 4069 (32%) had confirmed dengue positive by Enzyme Linked Immunosorbent Assay (ELISA). The ratio of male cases was higher than female, and in age group 21-35 year (47%). Seasonal trend showed a gradual increase in positivity from June with high peak in October. Circulation of all the four serotypes in area, higher monotypic infection by DENV-1 serotype (41%), followed by DENV-4, DENV-2 and DENV-3. Co-infection of different serotypes were also found. conclusion: The present study concluded that all four serotypes circulate with predominant being DENV-1 type and co-infection of different serotypes in the saurashtra region. Dengue mainly affected adult male population, and seasonal peak during monsoon and post-monsoon period.

scholarly journals Circulation of DENV-3 Genotype 3 during 2017 to 2018 in Delhi: A Single-Center Hospital-Based Study

2021 ◽  
Author(s):  
Abhishek Padhi ◽  
Ekta Gupta ◽  
Gaurav Singh ◽  
Shama Parveen ◽  
Arshi Islam ◽  
...  
Keyword(s):  
Acute Phase ◽  
Nonstructural Protein ◽  
Indian Subcontinent ◽  
Enzyme Linked Immunosorbent Assay ◽  
Care Hospital ◽  
Rt Pcr ◽  
Female Ratio ◽  
Genotype Iii ◽  
Nonstructural Protein 1 ◽  
New Variant

Abstract Introduction Delhi is hyperendemic for dengue virus (DENV) where all the four DENV have previously been reported. A constant vigilance of circulating DENV serotypes is important in surveillance, since the introduction of a new variant to areas affected by preexisting serotypes constitutes a risk factor for dengue hemorrhagic fever and dengue shock syndrome. Objectives This retrospective study was performed with an objective to determine the circulating serotype and genotype of DENV in acute phase blood samples of patients who have reported to a tertiary liver care hospital in New Delhi during the last 2 years (2017–2018). Methods The data of clinician-initiated testing for dengue nonstructural protein 1 (NS1) antigen (Ag) was searched in the institutional hospital information system. The serum sample of dengue NS1 Ag-positive cases confirmed by enzyme-linked immunosorbent assay (ELISA; PANBIO, Gyeonggi-do, ROK) and a fever duration of less than 5 days were retrieved from the laboratory archive. The DENV serotyping on these sample was performed by reverse transcriptase polymerase chain reaction (RT-PCR). Sequencing and phylogenetic analysis was done for the capsid premembrane (CprM) region to determine the genotype. Results A total of 440 acute-phase samples were received. Twenty one (4.77%) were positive for dengue NS1 Ag with a mean age of 35.1 years and male-to-female ratio of 1.1:1. Eight cases (38.09%) were positive by dengue RT-PCR and all belonged to DENV-3 serotypes. Phylogenetic tree analysis revealed DENV-3 clustered to genotype III with 100% homology with 2008 Indian subcontinent strain. Conclusion This study revealed circulation of DENV-3, genotype III in Delhi from 2017 to 2018, similar to the 2008 viral type. Virological surveillance is an important exercise to be done for viral infections with public threat and outbreak potential.

scholarly journals Epidemiological survey and screening strategy for dengue virus in blood donors from Yunnan Province

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Ling Li ◽  
Ying Li ◽  
Shaofang Lu ◽  
Jing Dong ◽  
Haixia Xu ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Southern China ◽  
Nonstructural Protein ◽  
Denv Infection ◽  
Screening Strategy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Donor Screening ◽  
Blood Center ◽  
Nonstructural Protein 1

Abstract Background Dengue virus (DENV) infection is increasingly common in southern China and can be transmitted through blood transfusion but is not currently part of donor screening throughout the region. We assessed DENV prevalence among donors at the Xishuangbanna Blood Center, Yunnan, to support development of DENV screening strategies. Methods Blood samples were collected randomly between June 2019 and August 2019. These were screened for anti-DENV IgG and IgM using enzyme-linked immunosorbent assay (ELISA). Then, all reactive samples and some randomly-chosen non-reactive samples were used to detect DENV RNAs using real-time polymerase-chain-reaction (RT-PCR) assays. After RT-PCR, samples were further tested for soluble nonstructural protein 1 (NS1) using the colloidal gold method. Donors demographics were also collected and assessed. Results Over the study period, 2254 donor samples were collected and tested for anti-DENV IgG and IgM by ELISA. This revealed 598 anti-DENV IgG and/or IgM reactive samples, a serological prevalence of 26.53%. Of these, 26 were RT-PCR positive and/or NS1 positive. Significant differences in DENV prevalence were noted by occupation (P = 0.001), education (P < 0.001), and ethnicity (P = 0.026). Conclusion The prevalence of DENV in Xishuangbanna Blood Center was higher than most other blood centers that have implemented DENV donor screening. Our study provides first-hand data about the prevalence of DENV and allows the development of a screening strategy for clinical use.

scholarly journals Epidemiological Survey and Screening Strategy for Dengue Virus in Blood Donors from Yunnan Province

2020 ◽  
Author(s):  
LING LI ◽  
YING LI ◽  
Shaofang Lu ◽  
Jing Dong ◽  
Haixia Xu ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Nonstructural Protein ◽  
Demographic Data ◽  
Screening Strategy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Blood Center ◽  
Nonstructural Protein 1 ◽  
Elisa Testing

Abstract BACKGROUNDDengue virus (DENV) can be transmitted through blood transfusion. DENV was not screened regularly in Xishuangbanna blood center. This study was conducted in Xishuangbanna blood center with an attempt to develop DENV screening strategies in one of China’s high-incidence areas.METHODSBlood samples were collected randomly between June 2019 and August 2019. These samples were first screened for anti-DENV IgG and IgM using enzyme-linked immunosorbent assay (ELISA). All reactive samples and some randomly-chosen non-reactive samples were used to detect DENV RNAs using real-time polymerase-chain-reaction (RT-PCR) assay. After RT-PCR assay, these samples were further tested for soluble nonstructural protein 1 (NS1) using the colloidal gold method. The demographic data of donors were collected.RESULTSA total of 2,254 donor samples were collected and tested for anti-DENV IgG and IgM by ELISA between June 2019 and August 2019. ELISA testing revealed that 598 donor samples were anti-DENV IgG and/or IgM reactive, with a serological prevalence of 26.53%. Among all the donor samples, 26 were RT-PCR positive and/or NS1 positive. Moreover, there were significant differences in the prevalence of DENV in terms of occupation (P=0.001), education(P<0.001) and ethnicity (P=0.026).CONCLUSIONThe prevalence of DENV in Xishuangbanna Blood Center was higher than most other blood centers that have implemented DENV donor screening. Our study provides the first-hand data about the prevalence of DENV and allows the development of a screening strategy for clinical use.

scholarly journals Epidemiological Survey and Screening Strategy for Dengue Virus in Blood Donors from Yunnan Province

2021 ◽  
Author(s):  
LING LI ◽  
YING LI ◽  
Shaofang Lu ◽  
Jing Dong ◽  
Haixia Xu ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Southern China ◽  
Nonstructural Protein ◽  
Denv Infection ◽  
Screening Strategy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Donor Screening ◽  
Blood Center ◽  
Nonstructural Protein 1

Abstract BACKGROUND Dengue virus (DENV) infection is increasingly common in southern China and can be transmitted through blood transfusion but is not currently part of donor screening throughout the region. We assessed DENV prevalence among donors at the Xishuangbanna Blood Center, Yunnan, to support development of DENV screening strategies.METHODS Blood samples were collected randomly between June 2019 and August 2019. These were screened for anti-DENV IgG and IgM using enzyme-linked immunosorbent assay (ELISA). Then, all reactive samples and some randomly-chosen non-reactive samples were used to detect DENV RNAs using real-time polymerase-chain-reaction (RT-PCR) assays. After RT-PCR, samples were further tested for soluble nonstructural protein 1 (NS1) using the colloidal gold method. Donors demographics were also collected and assessed.RESULTS Over the study period, 2,254 donor samples were collected and tested for anti-DENV IgG and IgM by ELISA. This revealed 598 anti-DENV IgG and/or IgM reactive samples, a serological prevalence of 26.53%. Of these, 26 were RT-PCR positive and/or NS1 positive. Significant differences in DENV prevalence were noted by occupation (P=0.001), education (P<0.001), and ethnicity (P=0.026).CONCLUSION The prevalence of DENV in Xishuangbanna Blood Center was higher than most other blood centers that have implemented DENV donor screening. Our study provides first-hand data about the prevalence of DENV and allows the development of a screening strategy for clinical use.

scholarly journals Development of an Antigen Capture Immunoassay Based on Monoclonal Antibodies Specific for Dengue Virus Serotype 2 Nonstructural Protein 1 for Early and Rapid Identification of Dengue Virus Serotype 2 Infections

2008 ◽  
Vol 16 (1) ◽  
pp. 88-95 ◽  
Author(s):  
Li-wen Qiu ◽  
Biao Di ◽  
Kun Wen ◽  
Xin-shuai Wang ◽  
Wei-hua Liang ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Nonstructural Protein ◽  
Rapid Identification ◽  
Serum Samples ◽  
Denv Serotypes ◽  
Nonstructural Protein 1 ◽  
Antigen Capture ◽  
Virus Serotype ◽  
Ns1 Antigen ◽  
Dengue Virus Serotype 2

ABSTRACT The dengue virus (DENV) has four distinct serotypes (DENV1, DENV2, DENV3, and DENV4) that require differentiation for effective prevention of morbid diseases. The recently developed DENV1-specific NS1 antigen capture enzyme-linked immunosorbent assay (ELISA) based on the monoclonal antibodies (MAbs) that recognize distinct epitopes on nonstructural protein 1 (NS1) of a specific DENV serotype is convenient and cost-effective, but assays have not yet been developed for DENV serotypes 2 to 4. This paper describes the development and validation of a DENV2-specific NS1 antigen capture ELISA by selection and optimization of the pair of well-characterized MAbs that recognized epitopes specific for DENV2 NS1 from a large panel of MAbs. The DENV2 NS1 ELISA displayed exclusive sensitivity with the DENV2 serotype and did not cross-react with the other three DENV serotypes. The sensitivity and specificity of the DENV2 NS1 ELISA were 83.3% (25/30) and 100% (504/504) when used to test 30 acute-phase serum samples from patients infected with DENV2 identified by virus isolation or reverse transcription-PCR serotyping and 504 serum samples from healthy individuals, respectively. The specificity of this assay was also evaluated using a panel of serum samples which were positive for DENV1, other flaviviruses, and nonflaviviruses; no cross-reactions were observed in these clinical samples. The DENV2 NS1 ELISA was eightfold more sensitive than a commercially available serotype-cross-reactive NS1 ELISA (Panbio Diagnostics, Brisbane, Australia) when the two assays were used to test the DENV2-infected cell culture supernatants in parallel. The Panbio NS1 ELISA displayed variation in sensitivity between DENV serotypes. The DENV2-specific NS1 antigen capture ELISA can be used as a tool for the rapid identification of DENV2 infections.

scholarly journals Full Serotype- and Group-Specific NS1 Capture Enzyme-Linked Immunosorbent Assay for Rapid Differential Diagnosis of Dengue Virus Infection

2011 ◽  
Vol 18 (3) ◽  
pp. 430-434 ◽  
Author(s):  
Xixia Ding ◽  
Dongmei Hu ◽  
Yue Chen ◽  
Biao Di ◽  
Jing Jin ◽  
...  
Keyword(s):  
Differential Diagnosis ◽  
Virus Infection ◽  
Dengue Virus ◽  
Nonstructural Protein ◽  
Dengue Infection ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dengue Virus Infection ◽  
Effective Prevention ◽  
Nonstructural Protein 1

ABSTRACTDengue virus (DENV), a member of theFlavivirusfamily, has four distinct serotypes (DENV serotype 1 [DENV1], DENV2, DENV3, and DENV4) that require differentiation for the effective prevention of morbid disease. Early and rapid differentiation between flaviviruses remains challenging. Full assays combining four individual, serotype-specific and one group-specific nonstructural protein 1 (NS1) antigen capture enzyme-linked immunosorbent assays (ELISAs) based on monoclonal antibodies (MAbs) against DENV NS1 were developed and validated. The sensitivities and specificities of the full NS1 ELISAs were evaluated with viral cultures and dengue acute-phase sera. Four serotype-specific NS1 ELISAs displayed high specificities for the detection and differentiation of appropriate serotypes. The group-specific NS1 ELISA was broadly reactive with the four dengue virus serotypes. None of the NS1 ELISAs displayed cross-reactivity with the other flaviviruses or samples from febrile patients with non-dengue virus infections. The full serotype- and group-specific MAb-based NS1 capture ELISAs may provide tools for the early detection and typing of dengue infection, which is preferable to reverse transcriptase PCR (RT-PCR) for the rapid differential diagnosis of dengue virus infection in the field.

scholarly journals Development of a novel NS1 competitive enzyme-linked immunosorbent assay for the early detection of Zika virus infection

PLoS ONE ◽  
2021 ◽  
Vol 16 (8) ◽  
pp. e0256220
Author(s):  
Julieta S. Roldán ◽  
Alejandro Cassola ◽  
Daniela S. Castillo
Keyword(s):  
Early Detection ◽  
Zika Virus ◽  
Nonstructural Protein ◽  
Control Strategies ◽  
Acute Infection ◽  
Diagnostic Methods ◽  
Congenital Defects ◽  
Enzyme Linked Immunosorbent Assay ◽  
Ns1 Protein ◽  
Nonstructural Protein 1

Zika virus (ZIKV) is a flavivirus that has emerged as a global health threat after the 2015 outbreak in the Americas, where devastating congenital defects were documented. There are currently no vaccines to prevent ZIKV infections nor commercially available clinical diagnostic tests demonstrated to identify ZIKV without cross-reactive interference of related flaviviruses. Early diagnosis is critical when treating symptomatic patients and in preventing ZIKV transmission. In this context, the development of sensitive and accurate diagnostic methods are urgently needed for the detection of ZIKV acute infection. The aim of this study consisted of obtaining monoclonal antibodies (mAbs) against denatured monomeric ZIKV Nonstructural protein 1 (ZNS1), a useful diagnostic marker for flavivirus early detection, in order to develop a highly specific and sensitive ZNS1 indirect competitive ELISA (icELISA). The production of hybridomas secreting ZNS1 mAbs was carried out through immunizations with denatured monomeric ZNS1. We selected 1F5 and 6E2 hybridoma clones, which recognized the heat-denatured ZNS1 hexameric form by indirect ELISA. Cross-reaction studies indicated that these mAbs specifically bind to a ZNS1 linear epitope, and that they do not cross-react with the NS1 protein from other related flaviviruses. The 1F5 mAb enabled the development of a sensitive and reproducible icELISA to detect and quantify small amounts of ZNS1 disease marker in heat-denatured human sera. Here, we establish a reliable 1F5 based-icELISA that constitutes a promising diagnostic tool for control strategies and the prevention of ZIKV propagation.

Sign in / Sign up

Export Citation Format

Share Document