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

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 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

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

Abstract BACKGROUND Dengue 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.METHODS Blood samples were collected randomly between June 2019 and August 2019. These samples were first screened for dengue IgG and IgM antibodies 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 colloidal gold method. The demographic data of DENV positive donors were collected.RESULTS A total of 2,254 donor samples were collected and tested for dengue IgG and IgM antibodies by ELISA between June 2019 and August 2019. ELISA testing revealed that 598 donor samples were anti-IgG and/or anti-IgM reactive, with a serological prevalence rate 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 rate of DENV in terms of 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 the first-hand data about the prevalence of DENV and allows development of a screening strategy for clinical use.

scholarly journals Comparison of Two Commercially Available Dengue Virus (DENV) NS1 Capture Enzyme-Linked Immunosorbent Assays Using a Single Clinical Sample for Diagnosis of Acute DENV Infection

2008 ◽  
Vol 15 (10) ◽  
pp. 1513-1518 ◽  
Author(s):  
Kovi Bessoff ◽  
Mark Delorey ◽  
Wellington Sun ◽  
Elizabeth Hunsperger
Keyword(s):  
Dengue Virus ◽  
Real Time ◽  
Virus Isolation ◽  
Nonstructural Protein ◽  
Clinical Sample ◽  
Denv Infection ◽  
Acute Stage ◽  
Rt Pcr ◽  
Nonstructural Protein 1 ◽  
Immunosorbent Assays

ABSTRACT Dengue virus (DENV) nonstructural protein 1 (NS1) has shown promise as a novel diagnostic marker of acute DENV infection. Current techniques used to diagnose acute DENV infection, including virus isolation and reverse transcription-PCR (RT-PCR), are costly and difficult to perform, while traditional serological assays have low sensitivities during the acute stage of infection. Two commercially available NS1 antigen capture enzyme-linked immunosorbent assays (ELISAs), the Platelia dengue NS1Ag test (Bio-Rad Laboratories, Marnes La Coquette, France) and the Pan-E dengue early ELISA test (Panbio Diagnostics, Brisbane, Australia), were evaluated against a well-characterized panel of 208 real-time RT-PCR- and virus isolation-positive sera, as well as 45 real-time RT-PCR- and serologically negative sera from patients with other acute febrile illnesses. The overall sensitivities were 64.9% (95% confidence interval [CI95], 58.2 to 71.1%) for the Panbio test and 83.2% (CI95, 77.5 to 87.7%) for the Bio-Rad test, with interserotype variation, especially for DENV serotype 4. Predictive models were constructed to identify factors that had a significant influence on a test's outcome with respect to this panel of samples in order to identify the conditions in which the test will be most effective as a diagnostic tool. The immunoglobulin G titer was found to be the only covariate that significantly influenced results in the Bio-Rad test, while serotype and the day postonset were found to significantly influence results in the Panbio test. We concluded that the NS1 capture ELISA is a useful tool that can improve testing algorithms to diagnose DENV infection in single samples from acute and early convalescent cases.

scholarly journals Potential Phosphorylation of Viral Nonstructural Protein 1 in Dengue Virus Infection

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1393
Author(s):  
Thanyaporn Dechtawewat ◽  
Sittiruk Roytrakul ◽  
Yodying Yingchutrakul ◽  
Sawanya Charoenlappanit ◽  
Bunpote Siridechadilok ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Nonstructural Protein ◽  
Antiviral Drug ◽  
Denv Infection ◽  
Site Directed Mutagenesis ◽  
Phosphorylation Sites ◽  
Post Translational Modification ◽  
Multifunctional Protein ◽  
Nonstructural Protein 1 ◽  
Underlying Mechanisms

Dengue virus (DENV) infection causes a spectrum of dengue diseases that have unclear underlying mechanisms. Nonstructural protein 1 (NS1) is a multifunctional protein of DENV that is involved in DENV infection and dengue pathogenesis. This study investigated the potential post-translational modification of DENV NS1 by phosphorylation following DENV infection. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), 24 potential phosphorylation sites were identified in both cell-associated and extracellular NS1 proteins from three different cell lines infected with DENV. Cell-free kinase assays also demonstrated kinase activity in purified preparations of DENV NS1 proteins. Further studies were conducted to determine the roles of specific phosphorylation sites on NS1 proteins by site-directed mutagenesis with alanine substitution. The T27A and Y32A mutations had a deleterious effect on DENV infectivity. The T29A, T230A, and S233A mutations significantly decreased the production of infectious DENV but did not affect relative levels of intracellular DENV NS1 expression or NS1 secretion. Only the T230A mutation led to a significant reduction of detectable DENV NS1 dimers in virus-infected cells; however, none of the mutations interfered with DENV NS1 oligomeric formation. These findings highlight the importance of DENV NS1 phosphorylation that may pave the way for future target-specific antiviral drug design.

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.

Anti-dengue virus nonstructural protein 1 antibodies contribute to platelet phagocytosis by macrophages

2016 ◽  
Vol 115 (03) ◽  
pp. 646-656 ◽  
Author(s):  
Ya-Ting Chu ◽  
Chiou-Feng Lin ◽  
Chih-Peng Chang ◽  
Trai-Ming Yeh ◽  
Robert Anderson ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Platelet Adhesion ◽  
Molecular Mimicry ◽  
Nonstructural Protein ◽  
Denv Infection ◽  
Dengue Disease ◽  
Nonstructural Protein 1 ◽  
Tnf Α ◽  
Β3 Integrin ◽  
Lines Of Evidence

SummaryThrombocytopenia is an important clinical manifestation of dengue disease. The hypotheses concerning the pathogenesis of thrombocytopenia include decreased production and increased destruction or consumption of platelets. We previously suggested a mechanism of molecular mimicry in which antibodies (Abs) directed against dengue virus (DENV) nonstructural protein 1 (NS1) cross-react with platelets. Furthermore, several lines of evidence show activation of endothelial cells (ECs) and macrophages are related to dengue disease severity. Previous studies also suggested that Ab-opsonised platelets facilitate the engulfment of platelets by macrophages. Here we show that TNF-α-activated ECs upregulate adhesion molecule expression to enhance the binding of platelets and macrophages and lead to anti-DENV NS1 Ab-mediated platelet phagocytosis. We further demonstrate that the interaction between macrophages and TNF-α-activated ECs requires binding of FcγR with the Fc region of platelet-bound anti-DENV NS1 Abs. Importantly, the binding of anti-DENV NS1 Abs to platelets did not interfere with platelet adhesion to ECs. The adhesion molecules ICAM-1 and β3 integrin expressed on ECs as well as the FcγR expressed on macrophages were critical in anti-DENV NS1 Ab-mediated platelet phagocytosis on activated ECs. Moreover, anti-DENV NS1 Abs dramatically enhanced platelet engulfment by macrophages in a murine model of DENV infection. Our study provides evidence for a novel role for anti-DENV NS1 Abs in the pathogenesis of thrombocytopenia in dengue disease by enhancing platelet phagocytosis by macrophages.

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 Comprehensive Evaluation of Differential Serodiagnosis between Zika and Dengue Viral Infections

2018 ◽  
Vol 57 (3) ◽  
Author(s):  
Day-Yu Chao ◽  
Matthew T. Whitney ◽  
Brent S. Davis ◽  
Freddy A. Medina ◽  
Jorge L. Munoz ◽  
...  
Keyword(s):  
Nonstructural Protein ◽  
Statistical Significance ◽  
Virus Titer ◽  
Denv Infection ◽  
Immunoglobulin M ◽  
Nucleic Acid Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Sample Collection ◽  
Serum Samples ◽  
Nonstructural Protein 1

ABSTRACT Diagnostic testing for Zika virus (ZIKV) or dengue virus (DENV) infection can be accomplished by a nucleic acid detection method; however, a negative result does not exclude infection due to the low virus titer during infection depending on the timing of sample collection. Therefore, a ZIKV- or DENV-specific serological assay is essential for the accurate diagnosis of patients and to mitigate potential severe health outcomes. A retrospective study design with dual approaches of collecting human serum samples for testing was developed. All serum samples were extensively evaluated by using both noninfectious wild-type (wt) virus-like particles (VLPs) and soluble nonstructural protein 1 (NS1) in the standard immunoglobulin M (IgM) antibody-capture enzyme-linked immunosorbent assay (MAC-ELISA). Both ZIKV-derived wt-VLP- and NS1-MAC-ELISAs were found to have similar sensitivities for detecting anti-premembrane/envelope and NS1 antibodies from ZIKV-infected patient sera, although lower cross-reactivity to DENV2/3-NS1 was observed. Furthermore, group cross-reactive (GR)-antibody-ablated homologous fusion peptide-mutated (FP)-VLPs consistently showed higher positive-to-negative values than homologous wt-VLPs. Therefore, we used DENV-2/3 and ZIKV FP-VLPs to develop a novel, serological algorithm for differentiating ZIKV from DENV infection. Overall, the sensitivity and specificity of the FP-VLP-MAC-ELISA and the NS1-MAC-ELISA were each higher than 80%, with no statistical significance. The accuracy can reach up to 95% with the combination of FP-VLP and NS1 assays. In comparison to current guidelines using neutralization tests to measure ZIKV antibody, this approach can facilitate laboratory screening for ZIKV infection, especially in regions where DENV infection is endemic and capacity for neutralization testing does not exist.

scholarly journals Serological Evaluation of Suspected West Nile Virus Human Cases following Its Introduction during a Dengue Outbreak in Puerto Rico in 2007

2011 ◽  
Vol 18 (6) ◽  
pp. 978-983 ◽  
Author(s):  
Elizabeth Hunsperger ◽  
Manuela Beltran ◽  
Luz Nereida Acosta ◽  
Jorge Jordan-Munoz ◽  
Jomil Torres ◽  
...  
Keyword(s):  
Puerto Rico ◽  
West Nile Virus ◽  
Nonstructural Protein ◽  
Diagnostic Testing ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Serum Samples ◽  
West Nile ◽  
Nonstructural Protein 1 ◽  
Testing Algorithm

ABSTRACTA laboratory testing algorithm was evaluated to confirm West Nile virus (WNV) infection in human serum following the introduction of the virus in Puerto Rico in 2007. This testing algorithm used two standard diagnostic assays, the IgM antibody capture enzyme-linked immunosorbent assay (MAC ELISA) and real-time reverse transcriptase PCR (RT-PCR), along with two nonconventional assays, the nonstructural protein 1 (NS1) ELISA and a 90%-plaque-reduction neutralization test (PRNT90) with IgG depletion for dengue virus (DENV) and WNV. A total of 2,321 serum samples from suspected WNV human cases were submitted for testing. Approximately one-third (867, 37%) were cross-reactive for DENV and WNV by MAC ELISA and had negative RT-PCR results for both viruses. Of a subset of 43 samples tested, 31 (72%) of these cases were identified as positive for DENV in the PRNT90with IgG depletion and 8 (19%) were positive in the DENV NS1 antigen ELISA. These two assays combined differentiated 36 (84%) of the samples that could not be diagnosed using the standard diagnostic testing methods.

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