scholarly journals Evaluation of VIDAS® diagnostic assay prototypes detecting dengue virus NS1 antigen and anti-dengue virus IgM and IgG antibodies

2021 ◽  
Author(s):  
Sompavanh Somlor ◽  
Ludovic Brossault ◽  
Marc Grandadam
Keyword(s):  
Dengue Virus ◽  
High Sensitivity ◽  
Denv Infection ◽  
Diagnostic Assay ◽  
Igg Antibodies ◽  
Rt Pcr ◽  
Ns1 Antigen ◽  
Comparable Performance ◽  
Adults And Children ◽  
Automated Platform

AbstractDengue is a tropical disease caused by the mosquito-borne dengue virus (DENV) and affecting an estimated 96 million people each year. Performant, rapid and easy-to-use assays are needed for the accurate diagnosis of acute DENV infection. This study evaluated the performance of three prototype assays developed for the VIDAS® automated platform to detect dengue NS1 antigen and anti-dengue IgM and IgG antibodies. Positive and negative agreement with competitor enzyme-linked immunosorbent assays (ELISA) and rapid diagnostic tests (RDT) was evaluated in 91 Lao patients (57 adults, 34 children) with acute DENV infection. The VIDAS® NS1 assay showed the best overall agreement (95.6%; 95% confidence interval [CI]: 89.1-98.8%) with the competitor NS1 ELISA (Focus Diagnostics). Both assays also demonstrated high sensitivity relative to DENV RNA real-time RT-PCR set as gold standard (85.7% [95% CI: 74.3-92.6%] and 83.9% [95% CI: 72.2-91.3%], respectively). Concordance of the VIDAS® results was overall higher with the competitor ELISA than with the RDT, suggesting a better performance of VIDAS® assays over RDT. The overall agreement of VIDAS® IgM and IgG assays with the competitor ELISA (Panbio/Abbott) was moderate (72.5%, 95% CI [62.6-80.6%] for IgM; 76.9%, 95% CI [67.3-84.4%] for IgG), highlighting differences in sensitivity and/or specificity between these assays. In all comparisons, test agreements did not differ significantly between adults and children, indicating comparable performance of the VIDAS® assays in both populations. Altogether, the VIDAS® dengue prototypes performed very well and appear to be suitable for routine detection of dengue NS1 antigen and anti-dengue IgM/IgG antibodies.

scholarly journals Evaluation of VIDAS® Diagnostic Assay Prototypes Detecting Dengue Virus NS1 Antigen and Anti-Dengue Virus IgM and IgG Antibodies

Diagnostics ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1228
Author(s):  
Somphavanh Somlor ◽  
Ludovic Brossault ◽  
Marc Grandadam
Keyword(s):  
Dengue Virus ◽  
High Sensitivity ◽  
Denv Infection ◽  
Diagnostic Assay ◽  
Igg Antibodies ◽  
Rt Pcr ◽  
Igm Elisa ◽  
Ns1 Antigen ◽  
Adults And Children ◽  
Automated Platform

Dengue is a serious tropical disease caused by the mosquito-borne dengue virus (DENV). Performant, rapid, and easy-to-use assays are needed for the accurate diagnosis of acute DENV infection. We evaluated the performance of three prototype assays developed for the VIDAS® automated platform to detect dengue NS1 antigen and anti-dengue IgM and IgG antibodies. Positive and negative agreement with competitor enzyme-linked immunosorbent assays (ELISA) and rapid diagnostic tests (RDT) was evaluated in 91 Lao patients (57 adults, 34 children) with acute DENV infection. The VIDAS® NS1 assay showed the best overall agreement (95.6%) with the competitor NS1 ELISA. Both VIDAS® NS1 and NS1 ELISA assays also demonstrated high sensitivity relative to DENV RNA RT-PCR set as gold standard (85.7% and 83.9%, respectively). In contrast, NS1 RDT was less sensitive relative to DENV RNA RT-PCR (72.7%). The overall agreement of VIDAS® IgM and IgG assays with the competitor assays was moderate (72.5% for IgM ELISA, 76.9% for IgG ELISA, and 68.7% for IgM and IgG RDT). In most analyses, test agreements of the VIDAS® assays were comparable in adults and children. Altogether, the VIDAS® dengue prototypes performed very well and appear to be suitable for routine detection of dengue NS1 antigen and anti-dengue IgM/IgG antibodies.

scholarly journals Dengue NS1 antigen kit shows high sensitivity for detection of recombinant dengue virus-2 NS1 antigen spiked with Aedes aegypti mosquitoes

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Philip Raj Abraham ◽  
Bharathy R ◽  
Pradeep Kumar N ◽  
Ashwani Kumar
Keyword(s):  
Public Health ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Infected Patient ◽  
Specific Treatment ◽  
High Sensitivity ◽  
Field Evaluation ◽  
Denv Infection ◽  
Ns1 Protein ◽  
Ns1 Antigen

AbstractDengue, caused by the dengue virus (DENV) is a significant vector-borne disease. In absence of a specific treatment and vaccine, dengue is becoming a rising threat to public health. Currently, control of dengue mainly focuses on the surveillance of the mosquito vectors. Improved surveillance methods for DENV in mosquito populations would be highly beneficial to the public health. However, current methods of DENV detection in mosquitoes requires specialized equipment and expensive reagents and highly trained personnel. As an alternative, commercially available dengue NS1 antigen ELISA kits could be used for detection of DENV infection in Aedes aegypti mosquitoes. In this study, we explored the utility of commercially available Dengue NS1 antigen kit (J. Mitra & Co. Pvt. Ltd) for the detection of recombinant dengue virus-2 (rDENV-2) NS1 protein and serum of dengue infected patient spiked with Ae. aegypti mosquito pools. The kit was found to be highly sensitive and specific towards detection of all serotypes of DENV. Further, it could detect as low as 750 femto gram rDENV-2 NS1 protein. It was also observed that rDENV-2 NS1 antigen spiked with blood-fed and unfed mosquito pools could be detected. In addition, the kit also detected dengue infected patient serum spiked with Ae. aegypti mosquito pools. Overall, the Dengue NS1 antigen kit displayed high sensitivity towards detection of recombinant as well as serum NS1 protein spiked with Ae. aegypti mosquito pools and could be considered for the dengue virus surveillance after a field evaluation in Ae. aegypti mosquitoes.

scholarly journals Seroprevalence of Asymptomatic Dengue Virus Infection and Its Antibodies Among Healthy/Eligible Saudi Blood Donors: Findings From Holy Makkah City

2017 ◽  
Vol 8 ◽  
pp. 1178122X1769126 ◽  
Author(s):  
Ahmed M Ashshi ◽  
Saad Alghamdi ◽  
Adel G El-Shemi ◽  
Sabir Almdani ◽  
Bassem Refaat ◽  
...  
Keyword(s):  
Saudi Arabia ◽  
Dengue Virus ◽  
Blood Donors ◽  
Denv Infection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibodies ◽  
Dengue Disease ◽  
High Prevalence ◽  
Ns1 Antigen ◽  
Makkah City

Background: Threat to blood transfusion–transmitted dengue virus (DENV) and its antibodies has recently emerged worldwide. Dengue fever is an endemic disease in Saudi Arabia, particularly in its Western region. The aim of this study was to estimate the seroprevalence of asymptomatic DENV infection and its antibodies among eligible Saudi blood donors. Methods: Serum samples from 910 healthy/eligible adult male Saudi blood donors, who reside in Holy Makkah City of Saudi Arabia, were collected between March 2015 and August 2016 and screened for the detection of DENV nonstructural protein 1 (NS1) antigen and anti-DENV IgM and IgG antibodies using commercial enzyme-linked immunosorbent assay kits (Panbio, Brisbane, QLD, Australia). Results: Among the tested donors, 48 (5.3%) were seropositive for DENV-NS1 antigen, whereas 50 (5.5%) and 354 (38.9%) were seropositive for anti-DENV IgM and IgG antibodies, respectively. Seropositivity for DENV-NS1 antigen and/or anti-DENV IgM antibody among the tested donors reflects their ongoing asymptomatic viremic infectious stage with DENV during their donation time, whereas high prevalence of anti-DENV IgG seropositivity reflects the high endemicity of dengue disease in this region of Saudi Arabia. Conclusions: These results show high prevalence of asymptomatic DENV infection and its antibodies among Saudi blood donors, raising the importance of establishing blood screening for dengue disease at different blood donation services and units in Saudi Arabia to improve the guarantee of blood transfusions and to control DENV dissemination.

scholarly journals NS-1 antigen positive Dengue Infection and molecular characterization of Dengue Viruses in a private Medical College Hospitalin Dhaka, Bangladesh

2018 ◽  
Vol 17 (4) ◽  
pp. 669-673
Author(s):  
Mahmuda Siddiqua ◽  
Ahmed Nawsher Alam ◽  
AKM Muraduzzaman ◽  
Tahmina Shirin
Keyword(s):  
Virus Infection ◽  
Dengue Virus ◽  
Dengue Infection ◽  
Medical Science ◽  
Cost Effective ◽  
Rt Pcr ◽  
Dengue Virus Infection ◽  
Medical College ◽  
Virus Serotype ◽  
Ns1 Antigen

Introduction: Detection of dengue virus infection as soon as possible is critical for management of dengue virus infected patients. Immuno-chromatographic (ICT) tests are easy, cost effective method for dengue virus antigen detection.The sensitivity and specificity of ICT should compare with a gold standard test like RT-PCR. Aim of this study was to compare two test methods (ICT and RT-PCR), observe dengue serotype and seasonal impact on dengue infection.Methodology & result: The patients of Ibn Sina Medical College Hospital from October 2015 to October 2017 were tested for dengue NS1 antigen by ICT method. Out of 3201 sample tested 32.39% were found positive and 89 of which were re-tested for RT-PCR for comparison. Eighty eight of 89 NS1 positive cases showed positive by RT-PCR method giving an accuracy of 98.87%. Among the RT-PCR positive cases 45 were further analyzed for serotype. DEN-1, DEN-2 or both DEN- 1 and DEN-2 were found in 21, 23 and 1cases respectively. No cases of DEN-3 or DEN-4 were detected.Conclusion: This study showed that easily available and cost effective dengue NS1 antigen detection method (ICT) is as effective as molecular test (RT-PCR). DEN-1 and DEN-2 serotype were prevalent during last few years in Bangladesh. Continuous monitoring of dengue virus serotype is important for prevention and control of sudden epidemic by other serotype. Alert to be more during post monsoon when the peak of dengue virus infection was observed.Bangladesh Journal of Medical Science Vol.17(4) 2018 p.669-673

scholarly journals Antiviral Efficacy of the Anesthetic Propofol against Dengue Virus Infection and Cellular Inflammation

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Ting-Jing Shen ◽  
Chia-Ling Chen ◽  
Ming-Kai Jhan ◽  
Po-Chun Tseng ◽  
Rahmat Dani Satria ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Cell Model ◽  
Antiviral Agents ◽  
Denv Infection ◽  
Severe Dengue ◽  
Cellular Inflammation ◽  
Antiviral Effects ◽  
Endosomal Acidification ◽  
Adults And Children

Propofol, 2,6-diisopropylphenol, is a short-acting intravenous sedative agent used in adults and children. Current studies show its various antimicrobial as well as anti-inflammatory effects. Dengue virus (DENV) is an emerging infectious pathogen transmitted by mosquitoes that causes mild dengue fever and progressive severe dengue diseases. In the absence of safe vaccines and antiviral agents, adjuvant treatments and supportive care are generally administered. This study investigated the antiviral effects of propofol against DENV infection and cellular inflammation by using an in vitro cell model. Treatment with propofol significantly inhibited DENV release 24 h postinfection in BHK-21 cells. Furthermore, it also blocked viral protein expression independent of the translational blockade. Propofol neither caused inhibitory effects on endosomal acidification nor prevented dsRNA replication. Either the proinflammatory TNF-α or the antiviral STAT1 signaling was reduced by propofol treatment. These results provide evidence to show the potential antiviral effects of the sedative propofol against DENV infection and cellular inflammation.

scholarly journals A Pan-Dengue Virus Reverse Transcription-Insulated Isothermal PCR Assay Intended for Point-of-Need Diagnosis of Dengue Virus Infection by Use of the POCKIT Nucleic Acid Analyzer

2016 ◽  
Vol 54 (6) ◽  
pp. 1528-1535 ◽  
Author(s):  
Yun Young Go ◽  
R. P. V. Jayanthe Rajapakse ◽  
Senanayake A. M. Kularatne ◽  
Pei-Yu Alison Lee ◽  
Keun Bon Ku ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Virus Infection ◽  
Dengue Virus ◽  
Rapid Test ◽  
Diagnostic Assay ◽  
Pcr Assay ◽  
Dengue Virus Infection ◽  
Qrt Pcr ◽  
Ns1 Antigen ◽  
Insulated Isothermal Pcr

Dengue virus (DENV) infection is considered a major public health problem in developing tropical countries where the virus is endemic and continues to cause major disease outbreaks every year. Here, we describe the development of a novel, inexpensive, and user-friendly diagnostic assay based on a reverse transcription-insulated isothermal PCR (RT-iiPCR) method for the detection of all four serotypes of DENV in clinical samples. The diagnostic performance of the newly established pan-DENV RT-iiPCR assay targeting a conserved 3′ untranslated region of the viral genome was evaluated. The limit of detection with a 95% confidence was estimated to be 10 copies ofin vitro-transcribed (IVT) RNA. Sensitivity analysis using RNA prepared from 10-fold serial dilutions of tissue culture fluid containing DENVs suggested that the RT-iiPCR assay was comparable to the multiplex real-time quantitative RT-PCR (qRT-PCR) assay for DENV-1, -3, and -4 detection but 10-fold less sensitive for DENV-2 detection. Subsequently, plasma collected from patients suspected of dengue virus infection (n= 220) and individuals not suspected of dengue virus infection (n= 45) were tested by the RT-iiPCR and compared to original test results using a DENV NS1 antigen rapid test and the qRT-PCR. The diagnostic agreement of the pan-DENV RT-iiPCR, NS1 antigen rapid test, and qRT-PCR tests was 93.9%, 84.5%, and 97.4%, respectively, compared to the composite reference results. This new RT-iiPCR assay along with the portable POCKIT nucleic acid analyzer could provide a highly reliable, sensitive, and specific point-of-need diagnostic assay for the diagnosis of DENV in clinics and hospitals in developing countries.

scholarly journals IDENTIFICATION OF DENGUE VIRUS SEROTYPES AT THE DR. SOETOMO HOSPITAL SURABAYA IN 2016 AND ITS CORRELATION WITH NS1 ANTIGEN DETECTION

2018 ◽  
Vol 23 (2) ◽  
pp. 151
Author(s):  
Jeine Stela Akualing ◽  
Aryati Aryati ◽  
Puspa Wardhani ◽  
Usman Hadi
Keyword(s):  
Polymerase Chain Reaction ◽  
Dengue Virus ◽  
Reverse Transcriptase ◽  
Ribonucleic Acid ◽  
Rna Virus ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Ns1 Antigen ◽  
Dengue Virus Serotypes ◽  
Polymerase Chain

Serotipe virus dengue yang beredar terus mengalami perubahan dan berbeda di setiap daerah. Pergeseran serotipe maupun genotipedi dalamnya, mempengaruhi terjadinya wabah dengue di berbagai negara. Perbedaan serotipe diduga bernasab dengan deteksi antigen(Ag) non-structural 1 (NS1), namun belum banyak penelitian yang mendukung hal tersebut. Penelitian potong lintang dikerjakan sejakFebruari-Agustus 2016 dan didapatkan 60 subjek infeksi virus dengue (IVD) dan 25 non-IVD. Ribonucleic acid (RNA) virus denguediperiksa di semua subjek menggunakan Simplexa Dengue Real-Time Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR)termasuk identifikasi serotipe virus dengue dan pemeriksaan NS1 menggunakan uji cepat NS1 Panbio. Perbedaan perbandingan variabelkategorikal dianalisis dengan uji Fisher Exact. Kenasaban antara serotipe dengan deteksi Ag NS1 dianalisis dengan Chi-Kuadrat. RNAvirus dengue terdeteksi di 43 dari 60 subjek IVD (71,7%). Serotipe terbanyak adalah DENV-3 (62,8%). Pergeseran dominasi serotipetelah terjadi di Surabaya, sebelumnya dari DENV-2 ke DENV-1 dan sekarang DENV-3, kemungkinan akibat mobilitas pejamu, transporvirus dan faktor geografis. Kepekaan uji cepat NS1 75% dan kekhasan 100%. Persentase deteksi NS1 antar serotipe berbeda bermakna(p=0,002). Deteksi NS1 lebih rendah pada DENV-1 dibandingkan DENV-2 (p=0,007) ataupun DENV-3 (p=0,003). Serotipe virusdengue bernasab dengan deteksi NS1 (p=0,005). Ciri serotipe maupun genotipe virus dengue kemungkinan mempengaruhi sekresiNS1. Telah terjadi pergeseran serotipe virus dengue di pasien IVD di Surabaya sehingga diperlukan surveillance berkesinambunganuntuk memperkirakan terjadinya wabah. Serotipe bernasab dengan deteksi NS1. Salah satu penyebab hasil negatif palsu NS1 adalahperbedaan serotipe.

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 Horseradish peroxidase-labeled rabbit anti-non-structural protein 1 of dengue virus-2 for the diagnosis of dengue virus infections

2019 ◽  
Vol 28 (2) ◽  
pp. 103-9
Author(s):  
Evy Suryani Arodes ◽  
Beti Ernawati Dewi ◽  
Tjahjani Mirawati Sudiro
Keyword(s):  
Early Diagnosis ◽  
Horseradish Peroxidase ◽  
Dengue Virus ◽  
Periodate Oxidation ◽  
Denv Infection ◽  
Negative Control ◽  
Enzyme Linked Immunosorbent Assay ◽  
Ns1 Protein ◽  
Structural Protein ◽  
Ns1 Antigen

BACKGROUND Early diagnosis of dengue virus (DENV) infection is essential for patient management and disease control. Detection of the antigen non-structural protein 1 (NS1) has been proven to provide early diagnosis of DENV infection. Thus, commercial NS1 antigen detection assays have been increasingly used and are becoming thetool of choice among clinicians to confirm DENV infection in Indonesia. METHODS To obtain anti-NS1 DENV antibody, NS1 protein (90 µg/ml) from the collection of the Department of Microbiology, Faculty of Medicine, Universitas Indonesia was injected into a rabbit. The anti-NS1 antibody from the rabbit was then labeled with horseradish peroxidase (HRP) using the periodate oxidation method. Sera were tested by enzyme-linked immunosorbent assay (ELISA) to detect NS1 from DENV-infected patients. RESULTS Serially diluted antibody labeled with HRP tested using the direct ELISA method showed the highest absorbance value at a 1:100 dilution (Mean [SD] = 1.35 [0.35]); even at a dilution as high as 1:3,200 (0.22 [0.15]), antibody labeled with HRP was able to detect the NS1 protein, although the absorbance value did not differ greatly from that of the negative control (0.13 [0.01]). CONCLUSIONS In an attempt to develop an NS1-based diagnostic test, polyclonal anti-NS1 DENV antibody was successfully produced as a diagnostic assay to determine the presence of DENV NS1 antigen in patients’ sera.

scholarly journals Nine year trends of dengue virus infection in Mumbai, Western India

2017 ◽  
Vol 9 (04) ◽  
pp. 296-302 ◽  
Author(s):  
Jayanthi Shastri ◽  
Manita Williamson ◽  
Nilima Vaidya ◽  
Sachee Agrawal ◽  
Om Shrivastav
Keyword(s):  
Virus Infection ◽  
Dengue Virus ◽  
Dengue Fever ◽  
Monsoon Season ◽  
Western India ◽  
Rt Pcr ◽  
Dengue Virus Infection ◽  
Virus Serotype ◽  
Wide Range ◽  
Ns1 Antigen

Abstract INTRODUCTION: Dengue virus (DENV) causes a wide range of diseases in humans, from acute febrile illness Dengue fever (DF) to life-threatening Dengue hemorrhagic fever (DHF) or Dengue shock syndrome (DSS). Factors believed to be responsible for spread of Dengue virus infection include explosive population growth, unplanned urban overpopulation with inadequate public health systems, poor standing water and vector control, climate changes and increased international recreational, business, military travel to endemic areas. All of these factors must be addressed to control the spread of Dengue and other mosquito-borne infections. The detection of Dengue virus RNA by reverse transcriptase PCR (RT-PCR) in human serum or plasma samples is highly indicative of acute Dengue fever. Moreover, the method is able to identify the Dengue virus serotype by demonstrating defined sequence homologies in the viral genomic RNA. METHODS AND RESULTS: During the nine year period of this study analysis, 6767 strongly suspected cases were tested by RT-PCR. 1685 (24.9%) were Dengue PCR positive and confirmed as Dengue cases. Observations on the seasonality were based on the nine year's data as the intensity of sampling was at its maximum during monsoon season. Dengue typing was done on 100 positive samples after storage of Dengue RNA at – 80°C. Dengue serotypes were detected in 69 samples of which Dengue 2 was most predominant. 576 samples were processed for NS1 antigen and PCR simultaneously. 19/576 were positive (3.3 %) for NS1 as well as by PCR. 23/576 samples were negative for NS1 antigen, but were positive by RT-PCR. The remaining 534 samples which were negative for NS1 antigen were also negative by Dengue RT-PCR. CONCLUSION: In this study we sought to standardize rapid, sensitive, and specific fluorogenic probe-based RT-PCR assay to screen and serotype a representative range of Dengue viruses that are found in and around Mumbai. Qualitative Dengue virus TaqMan assays could have tremendous utility for the epidemiological investigation of Dengue illness and especially for the study of the viremic response with candidate live-attenuated dengue virus vaccines.

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