Enhancing the sensitivity of Dengue virus serotype detection by RT-PCR among infected children in India

2017 ◽  
Vol 244 ◽  
pp. 46-54 ◽  
Author(s):  
Syed Fazil Ahamed ◽  
Rosario Vivek ◽  
Shalini Kotabagi ◽  
Kaustuv Nayak ◽  
Anmol Chandele ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Rt Pcr ◽  
Virus Serotype

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 Rapid Detection and Molecular Typing of Dengue Virus by Using Multiplex-Nested-RT-PCR

2015 ◽  
Vol 11 (2) ◽  
Author(s):  
Nastiti Wijayanti ◽  
Hera Nirwati ◽  
Tri Wibawa ◽  
Aris Haryanto ◽  
S. Sutaryo
Keyword(s):  
Dengue Virus ◽  
Nested Pcr ◽  
Genotypic Variation ◽  
Clinical Samples ◽  
Rt Pcr ◽  
Virus Serotype ◽  
Pcr Multiplex ◽  
Pcr Products ◽  
One Step ◽  
Nested Pcr Assays

world. We have evaluated the combination of one-step RT-PCR and multiplex nested PCR assays for detectingdengue viruses from clinical samples. Twelve patients were screened for the dengue virus, using a pair of primersthat conserve for several Flavivirus. The results showed that in 12 suspect patients, 100% were positive for Flavivirusand there are some genotypic variation among them, that indicated by several RT-PCR products higher than 511 bp,the expected product for RT-PCR. Further assay was performed to clarify the presence and serotypes of dengue virususing multiplex nested PCR. Serotyping results indicated that 83,3% of samples can be confirmed for dengue virus.Among the dengue virus positive 16,7 % are dengue-2, 16.7 % are dengue-3, and the most common 50% are dengue-4,whereas dengue-1 were not found among the patients. The combination of RT-PCR and multiplex nested PCR assaycan be used for rapid analysis dengue samples in early phase which is potentially useful for clinical, epidemiologyand also evolutionary studies.Key words: Flavivirus, dengue virus, serotype, RT-PCR, multiplex nested PCR

scholarly journals Dengue virus serotype 4 and chikungunya virus coinfection in a traveller returning from Luanda, Angola, January 2014

2014 ◽  
Vol 19 (10) ◽  
Author(s):  
R Parreira ◽  
S Centeno-Lima ◽  
A Lopes ◽  
D Portugal-Calisto ◽  
A Constantino ◽  
...  
Keyword(s):  
Virus Infection ◽  
Dengue Virus ◽  
Chikungunya Virus ◽  
Clinical Laboratory ◽  
Phylogenetic Analyses ◽  
Genomic Sequences ◽  
Rt Pcr ◽  
Virus Serotype ◽  
First Time

A concurrent dengue virus serotype 4 and chikungunya virus infection was detected in a woman in her early 50s returning to Portugal from Luanda, Angola, in January 2014. The clinical, laboratory and molecular findings, involving phylogenetic analyses of partial viral genomic sequences amplified by RT-PCR, are described. Although the circulation of both dengue and chikungunya viruses in Angola has been previously reported, to our knowledge this is the first time coinfection with both viruses has been detected there.

scholarly journals Intrinsic variation in the vertically transmitted insect-specific core virome of Aedes aegypti

2021 ◽  
Author(s):  
H. Coatsworth ◽  
J Bozic ◽  
J. Carrillo ◽  
E.A. Buckner ◽  
A.R. Rivers ◽  
...  
Keyword(s):  
United States ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Index Case ◽  
Natural Populations ◽  
The United States ◽  
Sample Type ◽  
Rt Pcr ◽  
Fundamental Information ◽  
Virus Serotype

AbstractSince 2009, local outbreaks of dengue (serotypes 1-3) mediated by Aedes aegypti mosquitoes have occurred in the United States, particularly in Florida (FL). In 2016 and 2017, dengue virus serotype 4 was found alongside several insect-specific viruses (ISVs) in pools of A. aegypti from sites in Manatee County, FL, in the absence of an index case. Although ISVs have been characterized in A. aegypti globally, the constitution of a core virome in natural populations remains unclear. Using mosquitoes sampled from the same area in 2018, we compared baseline ovary viromes of field (G0) and lab (Orlando) A. aegypti via metagenomic RNA sequencing. Across all samples, virome composition varied by sample type (field- or colony-derived). Four ISVs comprised >97% of virus sequences: a novel partiti-like virus (Partitiviridae), a previously described toti-like virus (Totiviridae), unclassified Riboviria, and four previously described orthomyxo-like viruses (Orthormyxoviridae). Whole or partial genomes for the toti-like virus, partiti-like virus, and one orthomyxo-like virus were assembled and analyzed phylogenetically. Multigenerational maintenance of these ISVs was confirmed orthogonally by RT-PCR in G0 and G7 mosquitoes, indicating vertical transmission as the mechanism for ISV sustentation. This study provides fundamental information regarding ISV ecology, persistence, and variation in A. aegypti in nature.

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.

scholarly journals IgA ANTI-DENGUE PROFILE IN SAMPLES WITH POSITIVE DENGUE PCR OR NS1

2019 ◽  
Vol 25 (1) ◽  
pp. 16
Author(s):  
M Thohirin Ramadhani ◽  
Aryati Aryati ◽  
M Vitanata Arfijanto
Keyword(s):  
Platelet Count ◽  
Dengue Virus ◽  
Diagnostic Marker ◽  
Clinical Manifestations ◽  
Rapid Test ◽  
Rt Pcr ◽  
Dengue Virus Infection ◽  
Diagnostic Instruments ◽  
Secondary Infections ◽  
Virus Serotype

Dengue Virus Infection (DVI) causes several clinical manifestations and requires varied diagnostic instruments. IgA anti-dengue as one of the diagnostic markers of DVI is suspected to have a shorter lifespan and greater sensitivity in detecting secondary infections compared to IgM anti-dengue. This study was conducted using 34 sera with positive RT-PCR or NS1 dengue virus. Samples were examined by a reverse flowimmunochromatographic method using AIM Dengue IgA Assure Rapid Test and will be analyzed its profile toward the day of fever, serotype, severity, platelet count, and type of infection. The overall sensitivity of IgA anti-dengue was 61.76% (n=34); in which IgA anti-dengue detected 14.29% primary and 66.67% secondary cases. IgA anti-dengue detected DEN1, DEN2, DEN3, and Mixed DEN1 – DEN3 virus serotype respectively 55.56%, 22.22%, 16.67%, and 5.56% (n=20). The day of fever was dominated by day-4 and day-5 respectively 28.57% (n=21). IgA anti-dengue was detected in DD, DHF grade I, II, and III 42,86%, 28.57%, 19.05%, and 9.52% (n=21) respectively. IgA anti-dengue detected in all levels of platelet count, it detected 60% in < 50,000 cell/mm3, 30% in 50,000 - 100.000 cell/mm3 and 10% in > 100,000 cell/mm3 platelet count sample (n=20). In conclusion, IgA anti-dengue showed a good performance, applicable as a diagnostic marker of DVI.

scholarly journals CORRELATION OF DENGUE VIRUS SEROTYPE AND DVI SEVERITY IN ADULT PATIENTS

2018 ◽  
Vol 24 (2) ◽  
pp. 185
Author(s):  
Suci Andriani ◽  
Aryati Aryati ◽  
Usman Hadi
Keyword(s):  
Virus Infection ◽  
Dengue Virus ◽  
Real Time ◽  
Dengue Hemorrhagic Fever ◽  
Hemorrhagic Fever ◽  
Adult Patients ◽  
Rt Pcr ◽  
Dengue Virus Infection ◽  
Virus Serotype ◽  
Degree Of Severity

The clinical manifestation of dengue virus infection is often not clear, varies widely from mild to severe. Exposure of dengue virus which serotype is different from a previous infection is a risk factor for the severe manifestation of dengue virus infection. Dengue hemorrhagic fever is classified into four degrees of severity based on clinical manifestations and laboratory results. Real-time RT-PCR Dengue can detect dengue virus serotype in early dengue virus infection. The aimed of this study was to prove the correlation between dengue virus serotype and degree of severity in adult patients. This study was a cross-sectional observational design done in February until July 2016. Subjects consisted of 100 dengue virus infection patients. Serum of the patients was examined using Real-time RT-PCR Dengue (Simplexa™ Dengue). It was shown that from 46 patients with DENV-3 serotype was 63%, DENV-2 serotype 17.4%, DENV-1 serotypes 17.4% and mixed infection of DENV-1 and DENV-3 serotype 2.2%. There was not any DENV-4 serotype. Dengue Hemorrhagic Fever (DHF) stage I was 47.8%, DHF stage II was 30.4%, DHF stage III was 10.9% and Dengue Fever was 10.9%. There was not any DHF stage IV. There was not enough evidence that DENV-3 correlated with the degree of severity (p= 0.510). Based on this research, DENV-3 serotype was the dominant serotype prevalent at the Dr. Soetomo Hospital. There was no correlation between viral dengue serotype and severity in dengue adult patients in this study. 

scholarly journals Micafungin Inhibits Dengue Virus Infection through the Disruption of Virus Binding, Entry, and Stability

2021 ◽  
Vol 14 (4) ◽  
pp. 338
Author(s):  
Yen-Chen Chen ◽  
Jeng-Wei Lu ◽  
Chia-Tsui Yeh ◽  
Te-Yu Lin ◽  
Feng-Cheng Liu ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Dengue Fever ◽  
Early Stage ◽  
Quantitative Polymerase Chain Reaction ◽  
Enterovirus 71 ◽  
Denv Infection ◽  
Immunofluorescence Assay ◽  
Dependent Manner ◽  
Virus Binding ◽  
Virus Serotype

Dengue fever is an arbovirus disease caused by infection with the dengue virus (DENV). Half of the world’s population lives under the threat of dengue fever, however, researchers have yet to develop any drugs that are clinically applicable to this infection. Micafungin is a member of the echinocandins family of anti-fungal drugs, capable of blocking the synthesis of β-1,3-D-glucan in the walls of fungal cells. Previous studies have demonstrated the effectiveness of Micafungin against infections of enterovirus 71 (EV71) and chikungunya virus (CHIKV). This is the first study demonstrating the effectiveness of micafungin in inhibiting the cytopathic effects of dengue virus serotype 2 (DENV-2) in a dose-dependent manner. Time-of-addition assays verified the inhibitory effects of micafungin in pre-treated, co-treated, and full-treatment groups. Binding and entry assays also demonstrated the effectiveness of micafungin in the early stage of DENV-2 infection. The virucidal efficacy of micafungin appears to lie in its ability to destroy the virion. Molecular docking assays revealed the binding of micafungin to the envelope protein of DENV-2, thereby revealing the mechanism by which micafungin affects the early stage of DENV infection and the stability of DENV. Two other micafungin analogs, caspofungin and anidulafungin, were also shown to have the antiviral effects on DENV-2. Finally, immunofluorescence assay (IFA) and reverse-transcription quantitative polymerase chain reaction (RT-qPCR) confirmed the broad anti-DENV ability of micafungin against dengue virus serotypes 1, 3, and 4 (DENV-1, DENV-3, and DENV-4). Taken together, these results demonstrate the potential of micafungin and its analogs as candidates for the development of broad-spectrum treatments for DENV infection.

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