Dengue virus infections: comparison of methods for diagnosing the acute disease

Poersch, Celina de Oliveira; Pavoni, Daniela Parada; Queiroz, Mário H.; Borba, Luana de; Goldenberg, Samuel; Santos, Claudia Nunes Duarte dos; Krieger, Marco Aurélio

doi:10.1016/j.jcv.2004.08.008

Cited by 59 publications

(27 citation statements)

References 20 publications

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“…This hypothesis is strengthened by data demonstrating NS1 and the circulating virus in the serum with the absence of immunoglobulins (9,14,12,20). The sensitivity of the test used in the hospital was fundamental to the early diagnosis of DENV infection in these patients.…”

Section: Discussionmentioning

confidence: 98%

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Dengue Virus 4 (DENV-4) Re-Emerges after 30 Years in Brazil: Cocirculation of DENV-2, DENV-3, and DENV-4 in Bahia

Campos

Pinho

Brandão³

et al. 2015

Jpn J Infect Dis

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SUMMARY: Dengue fever (DF) is a mosquito-borne viral disease of great concern in tropical and subtropical regions of the world. One important cause of the increase in DF is rapid development and urbanization has led to proliferation of the Aedes aegypti mosquito, the vector responsible for transmission of the illness. Surveillance of dengue virus (DENV) infection in Brazil shows the predominance of DENV-1, DENV-2, and DENV-3 until 2010. This study reports the reappearance of DENV-4 in Brazil for the first time in 30 years. Serum samples were collected from individuals (n ＝ 214) exhibiting fever and muscular pain in Bahia, Brazil, during 2011-2012. These samples were subjected to reverse transcription-polymerase chain reaction (RT-PCR)/nested PCR, which revealed that 82z of samples were positive for DENV-4; most were older age groups and exhibited a serological pattern consistent with a primary infection. The cocirculation of multiple DENV serotypes within the same city places the population at risk for a fatal form of the disease. Therefore, with the increasing incidence of severe DF cases, early diagnosis will be a priority for public health efforts in Brazil.

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Section: Discussionmentioning

confidence: 98%

“…With the increasing incidence of DENV infection, early diagnosis principally during the acute phase of the illness will be a public health priority in Brazil (9,10). Routine diagnostic test for DENV are based on the detection of NS1 viral protein and serum anti-DENV IgM/IgG (11,12).…”

Section: Introductionmentioning

confidence: 99%

Dengue Virus 4 (DENV-4) Re-Emerges after 30 Years in Brazil: Cocirculation of DENV-2, DENV-3, and DENV-4 in Bahia

Campos

Pinho

Brandão³

et al. 2015

Jpn J Infect Dis

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“…Molecular methods for detecting DEN nucleic acid include nested RT-PCR and real-time RT-PCR, in which the serotype is identified in separate reaction mixtures (1,20,22,30). The nested RT-PCR assay that is widely used in laboratories to identify DEN serotypes was found to be 10-fold more sensitive than the fourplex real-time RT-PCR assay (11,20,27). However, the fourplex real-time RT-PCR assay is faster than standard nested RT-PCR, as results from the real-time RT-PCR are available in approximately 3 h, compared to 10 h or more for the nested RT-PCR, which is the time required to run separate RT-PCR and PCRs and to visualize the PCR product by electrophoresis.…”

Section: Discussionmentioning

confidence: 99%

Serotype-Specific Detection of Dengue Viruses in a Fourplex Real-Time Reverse Transcriptase PCR Assay

2005

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The dengue (DEN) viruses are positive-strand RNA viruses in the genus Flavivirus. Dengue fever and dengue hemorrhagic fever/dengue shock syndrome are important human arboviral diseases caused by infection with one of four closely related but serologically distinct DEN viruses, designated DEN-1, DEN-2, DEN-3, and DEN-4 viruses. All four DEN serotypes are currently cocirculating throughout the subtropics and tropics, and genotypic variation occurs among isolates within a serotype. A real-time quantitative nucleic acid amplification assay has been developed to detect viral RNA of a single DEN virus serotype. Each primer-probe set is DEN serotype specific, yet detects all genotypes in a panel of 7 to 10 representative isolates of a serotype. In single reactions and in fourplex reactions (containing four primer-probe sets in a single reaction mixture), standard dilutions of virus equivalent to 0.002 PFU of DEN-2, DEN-3, and DEN-4 viruses were detected; the limit of detection of DEN-1 virus was 0.5 equivalent PFU. Singleplex and fourplex reactions were evaluated in a panel of 40 viremic serum specimens with 10 specimens per serotype, containing 0.002 to 6,000 equivalent PFU/ reaction (0.4 to 1.2 ؋ 10 6 PFU/ml). Viral RNA was detected in all viremic serum specimens in singleplex and fourplex reactions. Thus, this serotype-specific, fourplex real-time reverse transcriptase PCR nucleic acid detection assay can be used as a method for differential diagnosis of a specific DEN serotype in viremic dengue patients and as a tool for rapid identification and serotyping of DEN virus isolates.Dengue (DEN) viruses are a group of four closely related arboviruses in the genus Flavivirus, designated DEN-1, DEN-2, DEN-3, and DEN-4 viruses (7). Aedes aegypti is the primary mosquito vector of the DEN viruses, and humans are the primary host. As a result of the expansion of the range of A. aegypti in urban environments throughout the tropics and subtropics, transmission of DEN viruses has also increased, with an estimated 2.5 billion people at risk of infection (4, 7-9). As many as 100 million people per year may become infected, including approximately 400,000 cases of DEN hemorrhagic fever (DHF) and DEN shock syndrome (DSS), which are more serious manifestations often associated with secondary DEN virus infections (34). In areas of dengue hyperendemicity, all four of the DEN virus serotypes may be circulating simultaneously, and an increase in DHF and DSS has been reported in these areas (4, 7, 9, 10).The DEN viruses are closely related serologically; however, they are antigenically distinct, and primary infection by one DEN virus serotype does not protect against infection from another DEN virus serotype (8). The antibodies raised against the primary infecting DEN virus serotype may cross-react with the subsequent infecting DEN virus serotype and through opsomization may function to enhance the ability of the second DEN virus serotype to infect the host, in a process called antibody-dependent enhancement of infection (28). DHF and DSS...

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“…In contrast none of the other two DENV-3 viruses used induced any detectable viremia after plaque assay on Vero cells. We have analyzed vire- 1.4 1.9 2.2 2.9 2.8 2.5 1.8 0.9 8 mia for the 3 monkeys that received the DENV-3 74886 virus by real-time RT-PCR according to the methodology described by Poersch et al (2005). Monkey N44 showed 4 days of viremia whereas monkey O48 presented 1 day only and monkey M50 was not viremic by this assay either.…”

Section: Resultsmentioning

confidence: 99%

“…It can be argued that Vero cell plaque efficiency may differ among strains or serotypes and that the assays could have detected only a subpopulation not represented in the inoculum. In this regard the use of nucleic acid amplification procedures such as real-time PCR (Houng et al 2001, Poersch et al 2005 has been established for the quantification of DEN viral RNA in biological samples. Although it is true that measuring RNA copies may better reflect ongoing viral replication and would, therefore, be more appropriate to assess protective efficacy, the plaque assay reflects fully functional viral particles and not RNA molecules alone.…”

Section: Discussionmentioning

confidence: 99%