A simple one-step real-time RT-PCR for diagnosis of dengue virus infection

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.

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Imunocompetent Mice Model for Dengue Virus Infection

The Scientific World JOURNAL ◽

10.1100/2012/525947 ◽

2012 ◽

Vol 2012 ◽

pp. 1-12 ◽

Cited By ~ 11

Author(s):

Denise Gonçalves ◽

Rafael de Queiroz Prado ◽

Eric Almeida Xavier ◽

Natália Cristina de Oliveira ◽

Paulo Marcos da Matta Guedes ◽

...

Keyword(s):

Animal Model ◽

Virus Infection ◽

Dengue Virus ◽

Viral Disease ◽

Mice Model ◽

Rt Pcr ◽

Dengue Virus Infection ◽

Dengue Disease ◽

The Family ◽

The World

Dengue fever is a noncontagious infectious disease caused by dengue virus (DENV). DENV belongs to the familyFlaviviridae, genusFlavivirus, and is classified into four antigenically distinct serotypes: DENV-1, DENV-2, DENV-3, and DENV-4. The number of nations and people affected has increased steadily and today is considered the most widely spread arbovirus (arthropod-borne viral disease) in the world. The absence of an appropriate animal model for studying the disease has hindered the understanding of dengue pathogenesis. In our study, we have found that immunocompetent C57BL/6 mice infected intraperitoneally with DENV-1 presented some signs of dengue disease such as thrombocytopenia, spleen hemorrhage, liver damage, and increase in production of IFNγand TNFαcytokines. Moreover, the animals became viremic and the virus was detected in several organs by real-time RT-PCR. Thus, this animal model could be used to study mechanism of dengue virus infection, to test antiviral drugs, as well as to evaluate candidate vaccines.

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

Bangladesh Journal of Medical Science ◽

10.3329/bjms.v17i4.38334 ◽

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

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Spatio-temporal distribution analysis of circulating genotypes of dengue virus type 1 in western and southern states of India by a one-step real-time RT-PCR assay

Infection Genetics and Evolution ◽

10.1016/j.meegid.2019.103989 ◽

2019 ◽

Vol 75 ◽

pp. 103989 ◽

Cited By ~ 2

Author(s):

K. Alagarasu ◽

J.A. Patil ◽

M.B. Kakade ◽

A.M. More ◽

M. Bote ◽

...

Keyword(s):

Dengue Virus ◽

Real Time ◽

Temporal Distribution ◽

Distribution Analysis ◽

Rt Pcr ◽

Pcr Assay ◽

Southern States ◽

One Step ◽

Spatio Temporal

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One-step multiplex real-time RT-PCR for detection and typing of dengue virus

Molecular and Cellular Probes ◽

10.1016/j.mcp.2018.10.001 ◽

2019 ◽

Vol 43 ◽

pp. 86-91 ◽

Cited By ~ 4

Author(s):

Myung-Jin Mun ◽

Joon-Yong Bae ◽

Jin Hyuck Kim ◽

Soo Bok Kim ◽

Ilseob Lee ◽

...

Keyword(s):

Dengue Virus ◽

Real Time ◽

Rt Pcr ◽

One Step

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

Journal of Laboratory Physicians ◽

10.4103/jlp.jlp_169_16 ◽

2017 ◽

Vol 9 (04) ◽

pp. 296-302 ◽

Cited By ~ 5

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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Real time PCR. Application in dengue studies

Colombia Medica ◽

10.25100/cm.v42i2.778 ◽

2011 ◽

pp. 243-258 ◽

Cited By ~ 4

Author(s):

Jeanette Prada-Arismendy ◽

Jaime E Castellanos

Keyword(s):

Virus Infection ◽

Dengue Virus ◽

Real Time ◽

Real Time Pcr ◽

Dengue Virus Infection ◽

Virus Serotype ◽

Human Genome Sequencing ◽

Polymerase Chain ◽

Detection Of Mutations ◽

Dengue Virus Serotype 2

PCR (polymerase chain reaction) is a routinely used tool in every diagnostic and research laboratory. This technique has been used in detection of mutations and pathogens, forensic investigation, and even is the base tool for human genome sequencing. A modification of PCR technique, real time PCR, allows the quantification of nucleic acids with higher sensibility, specificity and reproducibility. This article is intended to clarify the foundations of real-time PCR, using an application model for virology. In the actual work, it was quantified the viral load of dengue virus serotype 2 produced from infected murine macrophages; the obtained results in this work established that murine strain BALB/c presents a greater susceptibility to dengue virus infection, which establishes BALB/c murine strain as a best model of study for investigation of dengue virus infection physiopathology.

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