Molecular techniques for the study and diagnosis of. Four denv serotypes 1, 2, 3, and 4 circulate in tropical and. These devices are gold coated silicon arrays with 20,000. Laboratory diagnosis 37 contamination, exposure to ambient temperatures for up to 7 days while in transit will not signi. Focus diagnostics innovative solutions for infectious. Clinical and laboratory diagnosis of dengue virus infection. Given that dengue virus infection elicits such a broad range of clinical symptoms, early and accurate laboratory diagnosis is essential for appropriate patient management. Early diagnosis of dengue, the most common mosquitoborne disease globally, remains challenging. With the spread of molecular testing and the development of new technologies and their opportunities, laboratory professionals and. Molecular techniques for the study and diagnosis of parasite.
A negative ns1 test result does not rule out infection. Current concepts in dengue pathogenesis pdf 92p this note describes about the global impact of dengue and its epidemiology, the clinical presentation of acute dengue and the pathogenesis of acute dengue infection. Dengue viruses are spread to people through the bite of an infected aedes species ae. Vaccines free fulltext efficient delivery of dengue. Tests have widely variable performance, depending on the manufacturer, circulating dengue types in a region, a patients past medical history, and duration of symptoms before presentation. Efficient and accurate diagnosis of dengue is of primary importance for clinical care i. Dengue virus infection presents a wide spectrum of manifestations including asymptomatic condition, dengue fever df, or severe forms, such as dengue hemorrhagic fever dhf and dengue shock syndrome dss in affected individuals. Several sensitive molecular diagnostics are currently deployed for the diagnosis of both denv and chikv, it is very difficult to delineate the two viral infections based on symptomology, as both share similar clinical presentation. Molecular diagnostics in dengue clinical research centre. Introduction dengue is the most rapidly spreading mosquito borne viral disease in the world.
This article highlights the clinical manifestations of dengue virus infection and the various molecular tests that were used for its laboratory. Dengue virus denv and chikungunya virus chikv are arboviruses that share the same aedes mosquito vectors and thus overlap in their endemic areas. Patients will be diagnosed sooner than before, and public health laboratories will have a clearer picture of the true number of dengue cases. Diagnostic tests based on detection of dengue virus denv genome are available with varying sensitivities and specificities. Performance of simplexa dengue molecular assay compared to. Antidengue virus denv immunoglobulin m igm seroconversion has been the reference standard for dengue diagnosis. It is transmitted via the bite of infected female aedes aegypti mosquito. In vitro diagnostics can detect diseases or other conditions, and can be. Dengue is common in more than 100 countries around the world. This report discusses the diagnostic tests used to confirm the presence of dengue in one individual. The median age was 28 years range 4 months to 68 years, 49% were males and 27% of patients were pediatric cases.
Dengue fever testing is used to determine whether a person with signs and symptoms and recent potential exposure has been infected with the dengue virus. Based on clinical criteria alone, zikv cannot be reliably distinguished from infections with other pathogens that cause an undifferentiated systemic febrile illness, including infections with two common arboviruses, dengue virus and. With the spread of molecular testing and the development of new technologies and their opportunities, laboratory professionals and physicians more than ever. Dgx have closed a purchase agreement under which diasorin has bought quests focus diagnostics, inc. Dengue fever is common in tropical areas, so take extra precautions if you travel somewhere with this type of climate. Performance of dengue diagnostic tests in a singlespecimen. Polymerase chain reaction pcr gel electrophoresis blotting techniques northern, southern gene expression analysis. The characteristics of denguepositive patients are summarized in table 2. Molecular methods in diagnosis of infectious diseases. Techniques in molecular biology to study the function of genes analysis of nucleic acids. Dengue presents the most uptodate overview of the field, encompassing various aspects of epidemiology, virus research, clinical management of the disease, pathogenesis, vector control and diagnosis. Dengue diagnosis is important for both management of patient as well as epidemiological surveillance in malaysia. Laboratory diagnosis of dengue viral infection learn.
Circulating levels of nonstructural glycoprotein 1 ns1 correlate with titres and increases in levels of specific immunoglobulin m igm, evidencing a recent acute infection with dengue virus. Fundamentals of molecular virology 2nd edition download fundamentals of molecular virology 2nd edition ebook pdf or read online books in pdf, epub, and mobi format. Instances of extreme disease severity are characterized by cytokine storm, life. Malathi postgraduate ii year department of microbiology chengalpattu medical college 2. Each year hundreds of people get infected with dengue in pakistan. Many laboratories now prefer to receive blood specimens in vials or tubes rather than blotted on paper, since the latter require special pretest processing. Lund, in progress in molecular biology and translational science, 2015. To improve particularly early laboratory diagnostics, a novel realtime rtpcr method was developed for the detection of denv14 rna based on taqman. Download pdf fundamentals of molecular virology 2nd. To assess the performance of the simplexa dengue assay, we performed comparison with.
Molecular diagnosis of dengue complements other existing assays antigen detection or serology useful for serotyping, measure viral load or diagnosis of fatal dengue cases increased demand for molecular testing for dengue. There are several virological and serological approaches to diagnose denv in blood with different sensitivities and specificities 3,57. The infection is difficult to diagnose without laboratory tests because symptoms may initially resemble those of other diseases, such as chikungunya infection. Each chapter contains an expert introduction to each subject that is next to technical details and many applications for molecular genetic testing that can be found in comprehensive reference lists at the end of each chapter. Handbook of diagnostic virology testing pdf 41p download book. The early prediction of severe dengue in patients without any warning signs who may later develop severe dhf is very. Laboratory diagnosis methods for confirming dengue virus infection may involve. Molecular diagnostics of infectious diseases harald. Dengue, laboratory test, diagnosis, assay introduction dengue, an arthropodborne disease is a major health concern especially in the tropics and subtropics. Rapid and accurate interpretation of dengue diagnostics in the context of dengue vaccination implementation. These two viruses also cause similar clinical presentations, especially in the initial stages of infection, with neither virus possessing any specific distinguishing clinical features. The clinical, serological and molecular diagnosis of emerging. Pdf dengue caused by four antigenically distinct serotype remains a.
Although the disease caused by denv is selflimiting in the first infection, the antibodydependent enhancement ade effect increases the mortality in the second infection with a heterotypic virus. Tedjo sasmono1 1eijkman institute for molecular biology, jakarta, indonesia, 2health polytechnic. Molecular detection and typing of dengue viruses from. Reprints and eprints sponsored supplements branded books journals career network. Clinical and laboratory diagnosis of dengue virus infection the. Jul 30, 2015 dengue virus epidemiology and clinical disease.
Molecular surveillance of dengue in semarang, indonesia revealed the circulation of an old genotype of dengue virus serotype1 sukmal fahri1,2,3. Newer detection tools that can fill this acknowledged gap in dengue diagnosis are urgently required. Strengthening position as worlds leading innovator in molecular diagnostics and development 031811. Rapid, lateralflow assays exist to detect the nonstructural protein 1 ns1 antigen and antidengue igm and igg.
Electrical detection of dengue biomarker using egg yolk. Of the 270 suspected patients, 65 were confirmed positive for acute zika virus infection by rrtpcr of serum, urine, or both. Since there is no specific efficient medicine in treatment, it is urgent. Handbook of diagnostic virology testing pdf 41p this note covers the following topics. In travelers dengue is rarely a lifethreatening disease, however in the current study, a fatality was documented in a young finnish patient who experienced a prolonged primary dengue infection. Towners presentation, one participant noted that the new molecular technologies reduce risk, so minimal containment levels are needed if pathogens such as ebola and marburg are indigenous to a particular locality. Presenting the latest molecular diagnostic techniques in one comprehensive volume the molecular diagnostics landscape has changed dramatically since the last edition of molecular microbiology. Introducing diasorin acquisition of focus diagnostics.
Molecular diagnostics tietz molecular diagnostics 5th molecular diagnostics laboratory molecular diagnostics 3rd edition pdf tietz textbook of clinical chemistry and molecular diagnostics tietz fundamentals of clinical chemistry and molecular diagnostics e book thm 180 diagnostics faq quest diagnostics laser diagnostics and optical measurement techniques chinese medicine study guide. For the clinical laboratorian, 2nd edition is the essential textbook of choice for anyone working in molecular diagnostics and who wants to remain current with this rapidly changing field. While significant efforts have been made to develop effective vaccines to mitigate this threat, the task has proven extremely challenging, with new approaches continually being sought. Molecular methods in diagnosis is a important topic to be understood by laboratory technology personal for implementation of emerging technologies for the grow. The assays were serotypespecific and did not crossdetect with other flaviviruses. The yellow book, centers for disease control and prevention. A global strategy on dengue fever and dhf was developed in 1995 and its implementation was bolstered in 1999. Concomitant circulation of dengue and chikungunya viruses has been sporadically reported and needs to be individually diagnosed. Since zika virus first appeared in french guiana, the nrc has investigated 270 samples collected 05 days after fever onset for molecular diagnosis of zika virus and dengue ns1. Early diagnosis is important for clinical screening, medical management, and disease surveillance. Molecular diagnosis of chikungunya virus chikv and dengue. Infection with any of the 4 dengue virus serotypes results in a diverse range of symptoms, from mild. Molecular technology molecular diagnosis is the most appropriate for infectious agents that are difficult to detect, identify, or test for susceptibility in a timely fashion with conventional mehtods.
Dengue virus is a currently a problematic global infection. Subsequently, the awareness of variable responses to the infection presenting. While advances in assay chemistry and detection continue to improve molecular diagnostics technology, blood samples are still collected using the 150yearold needlesyringe method. Focus diagnostics has been a leader in the development of infectious disease immunology assays for more than 25 years. Surface modified microprojection arrays for the selective. In vitro diagnostics are tests done on samples such as blood or tissue that have been taken from the human body. Rapid diagnostic tests for infectious diseases chapter. Molecular insight into dengue virus pathogenesis and its. Our dengue dxselect and dengue dx rapid tests reflect the tradition of quality, reliability and consistency expected by our customers around the world.
Techniques in molecular biology to study the function of. Dengue fever is an acute febrile arboviral disease affecting the tropical and subtropical regions of the world. Rapid diagnostic tests for infectious diseases chapter 11. Dengue and dengue hemorrhagic fever dhf are mosquitoborne viral disease caused by dengue virus, an arbovirus arthropod borne virus. Sayono3,5, suharyo hadisaputro3, edi dharmana3, din syafruddin1, r. Dengue fever is a virus transmitted to humans through bites from the aedes mosquito, a daybiting mosquito. Dengue infection can cause death of patients in its.
The simplexa dengue assay focus diagnostics is a newly developed realtime rtpcr method designed to detect and serotype denv simultaneously. Viewpoints and guidelines issued from an experts group consultation. Dengue virus is one of the most widespread mosquitoborne viruses worldwide, with an estimated annual infection rate around 390 million. S308 december 2010 with 2,038 reads how we measure reads. Current concepts in dengue pathogenesis pdf 92p download book. Our dengue dxselect and dengue dx rapid tests reflect the tradition of quality, reliability and consistency expected. If you think you have dengue pdf icon pdf 1 page see a healthcare provider if you develop a fever or have symptoms of dengue. Diagnostic techniques targeting viral biomarkers are generally positive in the. Hiv1 reservoirs and latency, molecular evolution of hepatitis viruses, metabolic aspects of hepatitis c virus infection, influenza c virus, viral vectors in neurobiology, avian and pandemic influenza. Molecular assays have comprehensively assisted in the diagnosis, treatment and epidemiological studies of parasitic diseases that affect people worldwide, helping to control parasitic disease mortality. Molecular diagnostics for the clinical laboratorian. Based on clinical criteria alone, zikv cannot be reliably distinguished from infections with other pathogens that cause an undifferentiated systemic febrile illness, including infections with two common. The incidence of this disease has increased over the last 50 years with 2. Dengue virus denv is a member of the flavivirus genus of singlestranded positivesense rna viruses that cause visceral and central nervous system disease in humans.
Jan 19, 2015 nonstructural protein 1 ns1 is secreted by dengue virus in the first days of infection and acts as an excellent dengue biomarker. Who works with governments and partners across the region to promote health, keep the world safe, and serve the vulnerable. Surface modified microprojection arrays have been developed as a novel platform for in vivo, needlefree biomarker capture. The spherical and enveloped dengue virus denv which belongs to. Molecular diagnosis based on reverse transcription rtpcr, such as onestep or nested rt. Molecular diagnostics of infectious diseases harald kessler. Molecular diagnostics current research and applications edited by.
It affects up to 100 million people each year, with 500,000 cases of dengue hemorrhagic fever dhf and dengue shock syndrome dss and. Laboratory diagnosis and diagnostic tests dengue ncbi. Currently, these assays are not typespecific, are expensive and are under evaluation for diagnostic accuracy and costeffectiveness in multiple settings. The growth of the global dengue virus diagnostic tests market is driven by an increase in the incidence of dengue. People with negative ns1 results should be tested for the presence of dengue igm antibodies to determine possible recent dengue exposure. Molecular diagnostics, third edition, focuses on the technologies and applications that professionals need to work in, develop, and manage a clinical diagnostic laboratory. The assays were evaluated with an external quality assessment panel from quality control for molecular diagnostics. Dengue is a rapidly spreading mosquitoborne viral disease. Other readers will always be interested in your opinion of the books youve read. Arboviruses molecular biology, evolution and control edited by.
Rapid dengue antigen detection tests can be used in field settings and provide results in less than an hour. Laboratory diagnosis method for confirming dengue viral infection involves use of one or combination of any of the following four different methods. Here, the direct electrical detection of ns1 from dengue type 2. Denv cycles in nature between its two principal mosquito vectors aedes albopictus or aedes aegypti and humans. Dengue virus rna or antigens were detected in 61 50% of 122 cases by rtpcr or ihc or both tests. Diagnostics free fulltext visual detection of dengue. Dengue viral rna detection detection of dengue viral nucleic acid by multiplex real time reverse transcriptase polymerase. Arboviruses molecular biology, evolution and control. Dengue diagnostics and dengue classificationcase management tdrirmdiagden05. Pdf dengue is an arthropodborne flavivirus that comprises four distinct serotypes den1, den2, den3 and den4 that constitute an antigenic. An estimated 50 million dengue infections occur annually. Several sensitive molecular diagnostics are currently deployed for the diagnosis of both denv and chikv, it is very difficult to delineate the two viral infections. Because the outcomes and management strategies for.
Molecular surveillance of dengue in semarang, indonesia. Rapid and accurate interpretation of dengue diagnostics in. This book covers various aspects of molecular virology. Dengue is now endemic in more than 100 countries, including pakistan. Diagnostic options and challenges for dengue and chikungunya. Zika virus zikv is an aedes mosquitoborne flavivirus that emerged in brazil in 2015 and then rapidly spread throughout the tropical and subtropical americas. Treat the symptoms of dengue and see your healthcare provider. According to who, dengue affects about 50100 million people worldwide, annually. Authoritative and stateoftheart, molecular diagnostics. Dengue virus, similar to wnv, is another example of a vectorborne acute flaviviral infection which can present with a spectrum of symptom severity and pathology. Dengue virus infection is a serious health problem infecting 2. In such a case, bsl3 and 4 laboratories are probably not needed, but some recipients seek highcontainment.
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