Dengue: a review of the laboratory tests a clinician must know to achieve a correct diagnosis.

Dengue is the most important disease caused by an arbovirus (1, 2, 3 and 4 serotypes) worldwide, especially in the tropical and sub-tropical regions. Its clinical manifestations range from asymptomatic infections to a severe disease characterized by hemorrhage and shock. The incidence of dengue virus activity in the Americas has substantially increased from 1980 to 1994. In Brazil, the increase in the incidence of dengue is especially linked to the dissemination of Aedes aegypti. Thus, a rapid and accurate dengue diagnosis is of paramount importance for effective control of dengue outbreaks [8]. Five serological tests have been used for the diagnosis of dengue infection: hemagglutination-inhibition (HI), complement fixation (CF), neutralization test (NT), immunoglobulin M (IgM) capture enzyme linked immunosorbent assay (MAC-ELISA) and indirect immunoglobulin G ELISA. The limitations of these techniques are the high cross-reactivity observed with these tests. Four methods of viral isolation have been routinely used for dengue viruses: intracerebral inoculation of newborn mice, inoculation on mammalian cell cultures, intrathoracic inoculation of adult mosquitoes, and inoculation on mosquito cell cultures. In recent years, several new diagnostic techniques have been developed and have proven very useful in dengue diagnosis, such as: nucleic and acid hybridization, RT-PCR. Currently, dengue diagnosis is based on serology, viral isolation and RNA detection. Enzyme-linked immunosorbent assays (ELISA) are still the most widely used technique for serological diagnosis, but they do not identify the dengue virus serotype responsible for the current infection, so molecular techniques may soon assume a very important role in dengue diagnosis. RT-PCR is definitely the most satisfactory test that can be used on these infections, since it has been shown to be able to detect dengue viruses up to the 10th day after the onset of the symptoms.

[1]  M. Tanaka,et al.  Rapid identification of flavivirus using the polymerase chain reaction. , 1993, Journal of virological methods.

[2]  S. Lam,et al.  Detection and serotyping of dengue viruses by PCR: a simple, rapid method for the isolation of viral RNA from infected mosquito larvae. , 1994, The Southeast Asian journal of tropical medicine and public health.

[3]  M. Varma,et al.  ISOLATION OF DENGUE VIRUSES IN MOSQUITO CELL CULTURES UNDER FIELD CONDITIONS , 1978, The Lancet.

[4]  G. Coelho,et al.  Dengue in Brazil: current situation and prevention and control activities. , 2002, Epidemiological bulletin.

[5]  S. Lam,et al.  Hemadsorption immunosorbent technique for the detection of dengue immunoglobulin M antibody , 1986, Journal of clinical microbiology.

[6]  R. Nogueira,et al.  Restriction enzyme analysis of American region dengue viruses , 2005, Archives of Virology.

[7]  J. Guesdon,et al.  Identification of dengue sequences by genomic amplification: rapid diagnosis of dengue virus serotypes in peripheral blood. , 1990, Journal of virological methods.

[8]  T. Cleary,et al.  Immunoglobulin G- and M-specific enzyme-linked immunosorbent assay for detection of dengue antibodies , 1979, Journal of clinical microbiology.

[9]  Sérgio Oliveira De Paula,et al.  Improved Detection of Dengue-1 Virus from IgM-Positive Serum Samples Using C6/36 Cell Cultures in Association with RT-PCR , 2003, Intervirology.

[10]  D. Gubler,et al.  Virological surveillance for dengue haemorrhagic fever in Indonesia using the mosquito inoculation technique. , 1979, Bulletin of the World Health Organization.

[11]  P. Mason,et al.  Flavivirus type-specific antigens produced from fusions of a portion of the E protein gene with the Escherichia coli trpE gene. , 1991, The American journal of tropical medicine and hygiene.

[12]  D. Gubler,et al.  The use of mosquitoes to detect and propagate dengue viruses. , 1974, The American journal of tropical medicine and hygiene.

[13]  G. Kuno,et al.  Mosquito cell cultures and specific monoclonal antibodies in surveillance for dengue viruses. , 1984, The American journal of tropical medicine and hygiene.

[14]  D. Gubler,et al.  Dengue and Dengue Hemorrhagic Fever , 1998, Clinical Microbiology Reviews.

[15]  V. Chow,et al.  Comparative analysis of NS3 sequences of temporally separated dengue 3 virus strains isolated from southeast Asia. , 1994, Intervirology.

[16]  J. Roux,et al.  Comparison of immunoglobulin G enzyme-linked immunosorbent assay (IgG-ELISA) and haemagglutination inhibition (HI) test for the detection of dengue antibodies. Prevalence of dengue IgG-ELISA antibodies in Tahiti. , 1989, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[17]  L. Figueiredo,et al.  An enzyme immunoassay for dengue antibody using infected cultured mosquito cells as antigen. , 1987, Journal of virological methods.

[18]  W. Hammon,et al.  New hemorrhagic fevers of children in the Philippines and Thailand. , 1960, Transactions of the Association of American Physicians.

[19]  R. Nogueira,et al.  Direct sequencing of genomic cDNA fragments amplified by the polymerase chain reaction for molecular epidemiology of dengue-2 viruses , 2005, Archives of Virology.

[20]  Y. Okuno,et al.  A modified PAP (peroxidase-anti-peroxidase) staining technique using sera from patients with dengue hemorrhagic fever (DHF): 4 step PAP staining technique. , 1979, Biken journal.

[21]  S. Hotta Experimental studies on dengue. I. Isolation, identification and modification of the virus. , 1952, The Journal of infectious diseases.

[22]  S. Devi,et al.  Detection of specific IgM in dengue infection. , 1987, The Southeast Asian journal of tropical medicine and public health.

[23]  R. Janeiro Genetic characterization of dengue virus type 3 isolates in the State of , 2002 .

[24]  A. Barrett,et al.  Rapid identification of flaviviruses based on conserved NS5 gene sequences. , 1993, Journal of virological methods.

[25]  A. Nisalak,et al.  An enzyme-linked immunosorbent assay to characterize dengue infections where dengue and Japanese encephalitis co-circulate. , 1989, The American journal of tropical medicine and hygiene.

[26]  L. Figueiredo,et al.  Enzyme immunoassay for the detection of dengue IgG and IgM antibodies using infected mosquito cells as antigen. , 1989, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[27]  T. Ksiazek,et al.  Viral hemorrhagic fevers , 1997 .

[28]  C. Peters,et al.  Demonstration of yellow fever and dengue antigens in formalin-fixed paraffin-embedded human liver by immunohistochemical analysis. , 1991, The American journal of tropical medicine and hygiene.

[29]  A. Nisalak,et al.  High circulating levels of the dengue virus nonstructural protein NS1 early in dengue illness correlate with the development of dengue hemorrhagic fever. , 2002, The Journal of infectious diseases.

[30]  R. Tesh A method for the isolation and identification of dengue viruses, using mosquito cell cultures. , 1979, The American journal of tropical medicine and hygiene.

[31]  H. Tan,et al.  The use of the single radial haemolysis technique in the serological diagnosis of dengue and Japanese encephalitis virus infections. , 1985, Bulletin of the World Health Organization.

[32]  B. A. Fonseca,et al.  The use of reverse transcription-polymerase chain reaction (RT-PCR) for the rapid detection and identification of dengue virus in an endemic region: a validation study. , 2002, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[33]  B. D. da Fonseca,et al.  Detection and identification of dengue‐1 virus in clinical samples by a nested‐PCR followed by restriction enzyme digestion of amplicons , 2002, Journal of medical virology.

[34]  A. Sabin Research on dengue during World War II. , 1952, The American journal of tropical medicine and hygiene.

[35]  P. J. Wright,et al.  Detection of flavivirus RNA in infected cells using photobiotin-labelled hybridization probes. , 1987, Journal of virological methods.

[36]  S. Kashima,et al.  Identification of Brazilian flaviviruses by a simplified reverse transcription-polymerase chain reaction method using Flavivirus universal primers. , 1998, The American journal of tropical medicine and hygiene.

[37]  G. Kuno,et al.  A PCR-restriction enzyme technique for determining dengue virus subgroups within serotypes. , 1994, Journal of virological methods.

[38]  S. Vázquez,et al.  Serological diagnosis of dengue by an ELISA inhibition method (EIM). , 1990, Memorias do Instituto Oswaldo Cruz.

[39]  D. Burke,et al.  Antibody Capture Immunoassay Detection of Japanese Encephalitis Virus Immunoglobulin M and G Antibodies in Cerebrospinal Fluid , 1982, Journal of clinical microbiology.

[40]  B. Miller,et al.  Detection of yellow fever viral RNA by nucleic acid hybridization and viral antigen by immunocytochemistry in fixed human liver. , 1989, The American journal of tropical medicine and hygiene.

[41]  R. Padmanabhan,et al.  Detection of dengue virus RNA using nucleic acid hybridization. , 1987, Journal of virological methods.

[42]  G. Kuno,et al.  Comparative sensitivity of three mosquito cell lines for isolation of dengue viruses. , 1985, Bulletin of the World Health Organization.

[43]  G. Kuno,et al.  An ELISA procedure for the diagnosis of dengue infections. , 1991, Journal of virological methods.

[44]  F. Pinheiro,et al.  Global situation of dengue and dengue haemorrhagic fever, and its emergence in the Americas. , 1997, World health statistics quarterly. Rapport trimestriel de statistiques sanitaires mondiales.

[45]  M. Cardosa,et al.  Development of a dot enzyme immunoassay for dengue 3: a sensitive method for the detection of antidengue antibodies. , 1988, Journal of Virological Methods.

[46]  B. D. da Fonseca,et al.  Comparison of techniques for extracting viral RNA from isolation-negative serum for dengue diagnosis by the polymerase chain reaction. , 2001, Journal of virological methods.

[47]  S. D. de Paula,et al.  Optimizing dengue diagnosis by RT-PCR in IgM-positive samples: comparison of whole blood, buffy-coat and serum as clinical samples. , 2002, Journal of virological methods.

[48]  S. Halstead,et al.  Pathogenesis of dengue: challenges to molecular biology. , 1988, Science.

[49]  Use of an inhibition method of ELISA in the serologic diagnosis of dengue. Preliminary report , 1989 .

[50]  M. Guzmán,et al.  Advances in dengue diagnosis , 1996, Clinical and diagnostic laboratory immunology.

[51]  L. Rosen,et al.  A simple technique for the detection of dengue antigen in mosquitoes by immunofluorescence. , 1977, The American journal of tropical medicine and hygiene.

[52]  M. C. Williams,et al.  Dengue in the Caribbean: virus isolation in a mosquito (Aedes pseudoscutellaris) cell line. , 1979, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[53]  G. Kuno,et al.  Use of 'original antigenic sin' theory to determine the serotypes of previous dengue infections. , 1993, Transactions of the Royal Society of Tropical Medicine and Hygiene.

[54]  A. Igarashi,et al.  Isolation of a Singh's Aedes albopictus cell clone sensitive to Dengue and Chikungunya viruses. , 1978, The Journal of general virology.

[55]  A. Nisalak,et al.  B cells are the principal circulating mononuclear cells infected by dengue virus. , 1999, The Southeast Asian journal of tropical medicine and public health.