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 Evaluation of Two New Commercial Tests for the Diagnosis of Acute Dengue Virus Infection Using NS1 Antigen Detection in Human Serum

2008 ◽  
Vol 2 (8) ◽  
pp. e280 ◽  
Author(s):  
Philippe Dussart ◽  
Laure Petit ◽  
Bhety Labeau ◽  
Laetitia Bremand ◽  
Alexandre Leduc ◽  
...  
Keyword(s):  
Virus Infection ◽  
Dengue Virus ◽  
Human Serum ◽  
Antigen Detection ◽  
Dengue Virus Infection ◽  
Ns1 Antigen

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 Clinical Evaluation of a Rapid Immunochromatographic Test for the Diagnosis of Dengue Virus Infection

1998 ◽  
Vol 5 (3) ◽  
pp. 407-409 ◽  
Author(s):  
Chew Theng Sang ◽  
Lim Siew Hoon ◽  
Andrea Cuzzubbo ◽  
Peter Devine
Keyword(s):  
Virus Infection ◽  
Dengue Virus ◽  
Secondary Infection ◽  
Rapid Test ◽  
Immunoglobulin M ◽  
Immunochromatographic Test ◽  
Dengue Virus Infection ◽  
Hemagglutination Inhibition Assay ◽  
Specific Immunoglobulin ◽  
Paired Serum

ABSTRACT A rapid immunochromatographic test was compared to the hemagglutination inhibition assay for separate determinations of dengue virus-specific immunoglobulin M (IgM) and IgG levels in paired serum specimens from 92 patients (34 with primary dengue virus infection, 35 with secondary dengue virus infection, and 23 without dengue virus infection). The rapid test showed 99% sensitivity in the diagnosis of dengue virus infection. The majority (30 of 34 [88%]) of patients with primary infection showed positive IgM but negative IgG, while 34 of 35 (97%) patients with secondary infection showed positive IgG with or without IgM. Specificity in nonflavivirus infections was 96% (1 of 23 positive). The rapid test should be a useful aid in rapid diagnosis of dengue virus infection.

Clinical evaluation of the NS1 antigen-capture ELISA for early diagnosis of dengue virus infection in Brazil

2010 ◽  
Vol 82 (8) ◽  
pp. 1400-1405 ◽  
Author(s):  
Luiza A. Castro-Jorge ◽  
Paula R.L. Machado ◽  
Camila A. Fávero ◽  
Marcos C. Borges ◽  
Luzia M.R. Passos ◽  
...  
Keyword(s):  
Virus Infection ◽  
Early Diagnosis ◽  
Dengue Virus ◽  
Clinical Evaluation ◽  
Dengue Virus Infection ◽  
Capture Elisa ◽  
Antigen Capture Elisa ◽  
Antigen Capture ◽  
Ns1 Antigen

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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