Serial real-time RT-PCR and serology measurements substantially improve Zika and Dengue virus infection classification in a co-circulation area

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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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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Laboratory Diagnosis of Dengue Virus Infection by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) and IgM-Capture Enzyme-Linked Immunosorbent Assay (ELISA)

Japanese Journal of Infectious Diseases ◽

10.7883/yoken.52.150 ◽

1999 ◽

Vol 52 (4) ◽

pp. 150-155

Author(s):

Ken-Ichiro Yamada ◽

Masaru Nawa ◽

Tomohiko Takasaki ◽

Sadao Yabe ◽

Ichiro Kurane

Keyword(s):

Polymerase Chain Reaction ◽

Virus Infection ◽

Dengue Virus ◽

Reverse Transcriptase ◽

Laboratory Diagnosis ◽

Enzyme Linked Immunosorbent Assay ◽

Rt Pcr ◽

Chain Reaction ◽

Dengue Virus Infection ◽

Polymerase Chain

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Serotype and Clinical Performance of Dengue Virus Infection on the Year 2009

Indonesian Journal of Tropical and Infectious Disease ◽

10.20473/ijtid.v1i2.2161 ◽

2010 ◽

Vol 1 (2) ◽

pp. 55 ◽

Cited By ~ 1

Author(s):

Soegeng Soegijanto ◽

Widodo Darmowandowo ◽

Amor Peraten Ginting ◽

Atsushi Yamanaka

Keyword(s):

Virus Infection ◽

Dengue Virus ◽

Clinical Manifestation ◽

Clinical Performance ◽

Secondary Infection ◽

Severe Disease ◽

Dengue Shock Syndrome ◽

Severe Dengue ◽

Rt Pcr ◽

Dengue Virus Infection

Dengue hemorrhagic fever is one of the important health problem in Indonesia, mortality rate is becoming decrease but many dengue shock syndrome cases is very difficult to be help. Previous study showed that some of DEN 2 and DEN 3 virus cases could show a clinical performance of severe dengue virus infection such as dengue shock syndrome. There are four serotype of dengue virus infection can cause primary and secondary infection. The aim of this research is to know the relationship between clinical performance of dengue virus infection and serotype dengue virus and also to know the role of primary and secondary infection and age of dengue virus cases. A prospective analytic observational study, which was conducted in Dr. Soetomo hospital since January 2009. RT-PCR was used to attempt to identify the infecting serotype from dengue virus isolated using vero cell. Antibody responses were measured by ELISA and clinical manifestation were measured with the WHO criteria 1997. Dengue serotype identification by RT-PCR was 70 patients. Virus types were DEN-2 65(92.8%), DEN-1 3(4.2%), and DEN-3 2(2.8%). Patients with DEN-1 genotype IV were more trend severe disease DSS and unusual infection. Commanly usually secondary exposure cause more severe clinical manifestation than primary exposure (p = 0.035) but in this study found that all of DEN-1 genotype IV, primary or secondary infection to show severe clinical manifestation of dengue virus infection. We can conclude that DEN-2 was the most dominant serotype in Dr. Soetomo Hospital. On Primary and secondary infection, DEN-1 genotype IV showing more severe than DEN-2 and DEN-3.

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EVALUASI PEMERIKSAAN IMUNOKROMATOGRAFI UNTUK MENDETEKSI ANTIBODI IgM DAN IgG DEMAM BERDARAH DENGUE ANAK

INDONESIAN JOURNAL OF CLINICAL PATHOLOGY AND MEDICAL LABORATORY ◽

10.24293/ijcpml.v12i2.850 ◽

2018 ◽

Vol 12 (2) ◽

pp. 88

Author(s):

Ety Retno Setyowati ◽

Aryati Aryati ◽

Prihatini Prihatini ◽

M.Y. Probohoesodo

Keyword(s):

Virus Infection ◽

Dengue Virus ◽

Diagnostic Value ◽

Rt Pcr ◽

Diagnostic Specificity ◽

Elisa Method ◽

Serum Samples ◽

Dengue Virus Infection ◽

Serological Tests ◽

Complex Technology

The gold standard diagnosis of DHF by RT-PCR needs a complex technology and is time consuming. Serological tests have beendeveloped to detect IgM and IgG anti dengue to determine primary as well as secondary acute phase infection. IgM and IgG antidenguetests by immunochromatography have been used, due to a high diagnostic validity, also because they are simple, practicable, easy, rapid(15–30 minutes), can be used in a single serum sample. ELISA method has been used as a confirmation method. The aim of this studyis to evaluate the immunochromatography method in detecting IgG and IgM anti dengue of DHF patients. The study was performedon 50 serum samples from patients of the ICU Department of Paediatrics Dr. Soetomo Hospital, Surabaya during July–August 2005with dengue virus infection according to the 1997, WHO criterion and 27 serum samples from non dengue virus infection patients.ELISA method showed positive infection in 44 samples. Immunochromatography method showed positive infection in 43 samples, butwas negative in 1 sample. Diagnostic sensitivity of Immunochromatography is 97.7% (43/44) and the diagnostic specificity is 92.6%(25/27). Immunochromatography method has a high diagnostic value in assisting the diagnosis of DHF.

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