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Molecular epidemiology of dengue 3 viruses and genetic relatedness among dengue 3 strains isolated from patients with mild or severe form of dengue fever in French Polynesia.

The nucleotide sequences of a short fragment of the envelope protein gene encoding amino acids 25 to 89 of 27 dengue 3 viruses were determined by direct sequencing of PCR-amplified products, and the viruses were compared regarding their time of isolation and geographic distribution. Four distinct genotypic groups were discerned at 6% divergence between nucleotide sequences. The first group contained isolates from the South Pacific (1988 to 1992), Singapore (1973) and Indonesia (1973 to 1991). The second group comprised viruses from Asia (1956 to 1989) including the reference strain H-87. The third was composed of one isolate from Thailand (1971), and the fourth included the early strains from French Polynesia (1964 to 1969) and from Puerto Rico (1963). Furthermore, the difference between early and recent strains from the South Pacific was as high as 12.3%. This observation suggests that the recent epidemics in the South Pacific were probably the consequence of the spread of a new variant that emerged from New Caledonia. However, relatedness between nucleotide sequence and disease severity, or between strains from epidemics with mild disease (New Caledonia) and strains from epidemics with severe disease (French Polynesia) could not be demonstrated.

Aedes↗

Evidence that maternal dengue antibodies are important in the development of dengue hemorrhagic fever in infants.

To establish the role of maternal dengue-specific antibodies in the development of dengue hemorrhagic fever and dengue shock syndrome caused by dengue 2 virus in infants, we examined sera from mothers of infants and toddlers with dengue hemorrhagic fever or dengue shock syndrome and mothers of infants with pyrexia of unknown origin. The mean titers of hemagglutination inhibition, neutralization, and infection-enhancing activities against dengue 2 virus were not statistically different among the three groups. However, among infants who developed dengue hemorrhagic fever/dengue shock syndrome there was a strong correlation between the mothers' dengue 2 neutralizing titers and infant age at the time of onset of severe illness, where no such correlation was found among the other two groups. Furthermore, the actual age at which dengue hemorrhagic fever/dengue shock syndrome occurred in each infant correlated with the age at which maximum enhancing activity for dengue 2 infection in mononuclear phagocytes was predicted. This critical time for the occurrence of dengue hemorrhagic fever/dengue shock syndrome was observed to be approximately 2 months after the time calculated for maternal dengue 2 neutralizing antibodies to degrade below a protective level. In addition, sera of mothers of infants with dengue hemorrhagic fever/dengue shock syndrome enhanced dengue 2 virus infection to a slightly greater degree than did sera from mothers of infants with pyrexia of unknown origin and toddlers with dengue hemorrhagic fever/dengue shock syndrome. These data are consistent with the hypothesis that maternal dengue antibodies play a dual role by first protecting and later increasing the risk of development of dengue hemorrhagic fever/dengue shock syndrome in infants who become infected by dengue 2 virus.

Age Factors↗

An information value based analysis of physical and climatic factors affecting dengue fever and dengue haemorrhagic fever incidence.

BACKGROUND: Vector-borne diseases are the most dreaded worldwide health problems. Although many campaigns against it have been conducted, Dengue Fever (DF) and Dengue Haemorrhagic Fever (DHF) are still the major health problems of Thailand. The reported number of dengue incidences in 1998 for the Thailand was 129,954, of which Sukhothai province alone reported alarming number of 682. It was the second largest epidemic outbreak of dengue after 1987. Government arranges the remedial facilities as and when dengue is reported. But, the best way to control is to prevent it from happening. This will be possible only when knowledge about the relationship of DF/DHF with climatic and physio-environmental agents is discovered. This paper explores empirical relationship of climatic factors rainfall, temperature and humidity with the DF/DHF incidences using multivariate regression analysis. Also, a GIS based methodology is proposed in this paper to explore the influence of physio-environmental factors on dengue incidences. Remotely sensed data provided important data about physical environment and have been used for many vector borne diseases. Information Values (IV) method was utilised to derive influence of various factors in the quantitative terms. Researchers have not applied this type of analysis for dengue earlier. Sukhothai province was selected for the case study as it had high number of dengue cases in 1998 and also due to its diverse physical setting with variety of land use/land cover types. RESULTS: Preliminary results demonstrated that physical factors derived from remotely sensed data could indicate variation in physical risk factors affecting DF/DHF. A composite analysis of these three factors with dengue incidences was carried out using multivariate regression analysis. Three empirical models ER-1, ER-2 and ER-3 were evaluated. It was found that these three factors have significant relation with DF/DHF incidences and can be related to the forecast expected number of dengue cases. The results have shown significantly high coefficient of determination if applied only for the rainy season using empirical relation-2 (ER-2). These results have shown further improvement once a concept of time lag of one month was applied using the ER-3 empirical relation. ER-3 model is most suitable for the Sukhothai province in predicting possible dengue incidence with 0.81 coefficient of determination. The spatial statistical relationship of various land use/land cover classes with dengue-affected areas was quantified in the form of information value received from GIS analysis. The highest information value was obtained for the Built-up area. This indicated that Built-up area has the maximum influence on the incidence of dengue. The other classes showing negative values indicate lesser influence on dengue epidemics. Agricultural areas have yielded moderate risk areas based on their medium high information values. Water bodies have shown significant information value for DF/DHF only in one district. Interestingly, forest had shown no influence on DF/DHF. CONCLUSION: This paper explores the potential of remotely sensed data and GIS technology to analyze the spatial factors affecting DF/DHF epidemic. Three empirical models were evaluated. It was found that Empirical Relatrion-3 (ER-3) has yielded very high coefficient of determination to forecast the number of DF/DHF incidence. An analysis of physio-environmental factors such as land use/land cover types with dengue incidence was carried out. Influence of these factors was obtained in quantitative terms using Information Value method in the GIS environment. It was found that built-up areas have highest influence and constitute the highest risk zones. Forest areas have no influence on DF/DHF epidemic. Agricultural areas have moderate risk in DF/DHF incidences. Finally the dengue risk map of the Sukhothai province was developed using Information Value method. Dengue risk map can be used by the Public Health Department as a base map for applying preventive measures to control the dengue outbreak. Public Health Department can initiate their effort once the ER-3 predicts a possibility of significant high dengue incidence. This will help in focussing the preventive measures being applied on priority in very high and high-risk zones and help in saving time and money.

Journal Article↗

Analysis of a recombinant dengue-2 virus-dengue-3 virus hybrid envelope protein expressed in a secretory baculovirus system.

In a step towards a tetravalent dengue virus subunit vaccine which is economical to produce, highly immunogenic and stable, a hybrid dengue virus envelope (E) protein molecule has been constructed. It consists of 36 amino acids from the membrane protein, the N-terminal 288 amino acids of the dengue-2 virus E protein plus amino acids 289-424 of the dengue-3 virus E protein. It has been engineered for secretory expression by fusion to a mellitin secretory signal sequence and truncation of the hydrophobic transmembrane segment. Using the baculovirus expression system and serum-free conditions, more than 95% of recombinant dengue-2 virus-dengue-3 virus hybrid E protein (rD2D3E) was secreted into the cell culture supernatant in a stable form with multiple features indicative of preserved conformation. The hybrid molecule reacted with a panel of dengue virus- and flavivirus-specific MAbs which recognize linear or conformational epitopes on dengue virions. Human dengue virus-specific antisera also reacted with the protein. The hybrid rD2D3E protein was able to inhibit the in vitro binding of dengue-2 and dengue-3 viruses to human myelomonocytic cells, suggesting that the receptor-binding epitope(s) was preserved. Adjuvant-free immunization with the hybrid protein induced an antibody response to both dengue-2 and dengue-3 virus in outbred mice, comparable in strength to that of individual rD2E and rD3E proteins. Notably, these antibody responses were primarily of the IgG2a and IgG2b isotype. A strong dengue virus cross-reactive T cell response was also induced in the mice, suggesting that dengue virus hybrid E proteins could form the basis of an efficacious multivalent dengue virus vaccine.

Animals↗

Evaluation of a commercial dengue NS1 antigen-capture ELISA for laboratory diagnosis of acute dengue virus infection.

A commercial dengue NS1 antigen-capture ELISA was evaluated to demonstrate its potential application for early laboratory diagnosis of acute dengue virus infection. Dengue virus NS1 antigen was detected in 199 of 213 acute serum samples from patients with laboratory confirmation of acute dengue virus infection but none of the 354 healthy blood donors' serum specimens. The dengue NS1 antigen-capture ELISA gave an overall sensitivity of 93.4% (199/213) and a specificity of 100% (354/354). The sensitivity was significantly higher in acute primary dengue (97.3%) than in acute secondary dengue (70.0%). The positive predictive value of the dengue NS1 antigen-capture ELISA was 100% and negative predictive value was 97.3%. Comparatively, virus isolation gave an overall positive isolation rate of 68.1% with a positive isolation rate of 73.9 and 31.0% for acute primary dengue and acute secondary dengue, respectively. Molecular detection of dengue RNA by RT-PCR gave an overall positive detection rate of 66.7% with a detection rate of 65.2 and 75.9% for acute primary dengue and acute secondary dengue, respectively. The results indicate that the commercial dengue NS1 antigen-capture ELISA may be superior to virus isolation and RT-PCR for the laboratory diagnosis of acute dengue infection based on a single serum sample.

Acute Disease↗

Dengue virus-specific human CD4+ T-lymphocyte responses in a recipient of an experimental live-attenuated dengue virus type 1 vaccine: bulk culture proliferation, clonal analysis, and precursor frequency determination.

We analyzed the CD4+ T-lymphocyte responses to dengue, West Nile, and yellow fever viruses 4 months after immunization of a volunteer with an experimental live-attenuated dengue virus type 1 vaccine (DEN-1 45AZ5). We examined bulk culture proliferation to noninfectious antigens, determined the precursor frequency of specific CD4+ T cells by limiting dilution, and established and analyzed CD4+ T-cell clones. Bulk culture proliferation was predominantly dengue virus type 1 specific with a lesser degree of cross-reactive responses to other dengue virus serotypes, West Nile virus, and yellow fever virus. Precursor frequency determination by limiting dilution in the presence of noninfectious dengue virus antigens revealed a frequency of antigen-reactive cells of 1 in 1,686 peripheral blood mononuclear cells (PBMC) for dengue virus type 1, 1 in 9,870 PBMC for dengue virus type 3, 1 in 14,053 PBMC for dengue virus type 2, and 1 in 17,690 PBMC for dengue virus type 4. Seventeen CD4+ T-cell clones were then established by using infectious dengue virus type 1 as antigen. Two patterns of dengue virus specificity were found in these clones. Thirteen clones were dengue virus type 1 specific, and four clones recognized both dengue virus types 1 and 3. Analysis of human leukocyte antigen (HLA) restriction revealed that five clones are HLA-DRw52 restricted, one clone is HLA-DP3 restricted, and one clone is HLA-DP4 restricted. These results indicate that in this individual, the CD4+ T-lymphocyte responses to immunization with live-attenuated dengue virus type 1 vaccine are predominantly serotype specific and suggest that a multivalent vaccine may be necessary to elicit strong serotype-cross-reactive CD4+ T-lymphocyte responses in such individuals.

Adult↗

Field- and laboratory-based active dengue surveillance in Chennai, Tamil Nadu, India: observations before and during the 2001 dengue epidemic.

BACKGROUND: Dengue cases are reported every year in the city of Chennai, Tamil Nadu, India. Since April 2001, longitudinal field- and laboratory-based active dengue surveillance has been carried out in Chennai to study dengue trends. METHOD: A serologic survey of people in Chennai using the hemagglutination inhibition test (HIT) was performed to determine evidence of prior exposure to dengue virus infections. Dengue virus infections and their serotypes were demonstrated in vectors. The serum samples from clinical dengue patients were analyzed for dengue virus-specific immunoglobulins M (IgM) and G (IgG) antibodies by 2 commercial ELISA kits. RESULTS: There was an increase in the percentage of children with monotypic antibody responses to dengue in the later survey (April 2.2%, September 9.93%). DEN-3 serotype infections were demonstrated in male Aedes aegypti mosquitoes collected in September 2001. Dengue virus infection was diagnosed in 74.5% (143/192) of cases. While dengue-specific IgM responses were predominant among infants with dengue fever, IgG and mixed responses (M + G) were seen in 85% of the children with severe forms of dengue. CONCLUSION: The findings from these investigations suggest that antibody surveys in children and virus detection in vectors may be included as early warning system parameters in laboratory-based proactive dengue surveillance.

Adolescent↗

Detection of dengue virus replication in peripheral blood mononuclear cells from dengue virus type 2-infected patients by a reverse transcription-real-time PCR assay.

While dengue virus is thought to replicate in mononuclear phagocytic cells in vivo, attempts to detect it in peripheral blood mononuclear cells (PBMC) by virus isolation or antigen detection have had variable and generally low rates. In this study, we developed a reverse transcription (RT)-real-time PCR assay to quantify positive- and negative-sense RNA of dengue virus type 2 within the cells. The assay includes an RT step using either sense or antisense primer followed by a real-time PCR step using the designed primers and probe, which target a capsid region highly conserved in dengue virus type 2 strains. It can be used to monitor the dynamic change of intracellular dengue virus RNA species during the course of infection. When this assay is employed in quantification of dengue virus RNA species in PBMC from 10 patients infected with dengue virus type 2, both positive- and negative-sense dengue RNA can be detected, indicating that dengue virus is actively replicating in PBMC in vivo. Moreover, the amounts of negative-sense dengue virus RNA in PBMC correlate very well with the viral load of dengue virus in plasma, suggesting that quantification of negative-sense dengue virus RNA in PBMC may provide another indicator of dengue virus replication in vivo. Use of this convenient, sensitive, and accurate method of quantification in clinical samples from patients with different disease severity would further our understanding of the pathogenesis of dengue.

Animals↗

[The specific epidemiological surveillance of dengue: the method and its importance since the dengue-2 epidemic in French Polynesia in 1996].

Dengue fever is present in tropical and subtropical regions and its geographical extension and the simultaneous increase of its mortality are worrisome. In endemic or epidemic countries, the aim of dengue-specific epidemiological surveillance is to confirm as soon as possible the circulation of a new viral dengue serotype, i.e. the beginning of an epidemic. The efficiency of the control strategy is improved by an earlier epidemic alert. In French Polynesia, dengue-3 virus circulated since 1989 at low level and, in May 1996, a specific epidemiological surveillance was undertaken because of the threat of a dengue-4 epidemic. From each suspected dengue case reported by 18 Polynesian physicians located in the Société Islands, a blood sample was taken for virological assay and clinical data were reported. Between May and November 1996, the virology unit of the Institut Malardé isolated 21 viruses (2 dengue-3 and 19 dengue-2) from 302 suspected cases. The dengue-specific epidemiological surveillance confirmed that dengue-2 virus was circulating and reduced the time of the epidemiological alert by 2 or 3 months compared to previous epidemics. Taking into account the day of illness, a logistic regression undertaken on the clinical data showed that the absence of cough was the only predictive sign of dengue diagnosis. The performance of this dengue-specific epidemiological surveillance system led us to consider its implementation in all concerned countries. A collaboration with international reference laboratories could be a solution for the developing countries.

Adolescent↗

Outbreak of dengue in Mumbai and predictive markers for dengue shock syndrome.

An alarming rise of dengue has been seen in Mumbai during the post-monsoon season. We undertook this prospective study in the pediatric wards and pediatric intensive care unit of B. J. Wadia Hospital for Children between 27 August 2003 and 10 October 2003 to determine the clinical features, laboratory abnormalities, and outcome of children affected with dengue and to determine the predictive markers for dengue shock syndrome. Fifty-one suspected dengue cases were tested for positivity of dengue by determination of dengue IgM antibodies by ELISA test. These positive cases were analysed for common clinical features, laboratory derangements, and outcome. Patients were subdivided into three subgroups: dengue fever (DF), dengue hemorrhagic fever (DHF), and dengue shock syndrome (DSS) as per WHO classification. Predictive markers for DSS were also determined. Thirty-nine patients had a positive dengue IgM titre, 20 patients had DHF, 18 patients had DSS, and one patient had DF The mean age of presentation was 4.9 years. Fever, hepatomegaly, vomiting, bleeding tendencies, erythematous rash, thrombocytopenia, elevated liver enzymes, and deranged PT and PTT were the predominant clinical and laboratory features. Predictive markers for DSS were younger age at onset, altered sensorium, paralytic ileus, and significantly deranged PT. Patients with DSS also had a longer recovery period and required more supportive management in the form of component therapy and ionotropic support. All three patients who died belonged to the DSS subgroup with case fatality rate for DSS being 16.6 per cent. None of the patients in the DHF or DF subgroup died. Endemicity of dengue fever is on the rise in Mumbai with increased incidence among children. Appropriate investigations, strict monitoring and prompt supportive management can reduce mortality in dengue. Predictive markers of DSS can reduce the mortality if promptly treated. Also prevention of transmission by mosquito control and maintaining water sanitation is required to effectively control this epidemic.

Analysis of Variance↗

[Detection of dengue virus genome RNA in some kinds of animals caught from dengue fever endemic areas in Hainan Island with reverse transcription-polymerase chain reaction].

Detection of dengue virus genome RNA in brain of bats caught from dengue fever endemic areas in Hainan Island were carried out with reverse transcription-polymerase chain reaction(RT-PCR). Positive result was demonstrated in 20 of 35 bats tested, with a positive rate of 57.14%; RT-PCR was applied for the detection of dengue virus genome RNA in sera of bats caught from the endemic areas in Hainan Island, 3(16.66%) of 18 sera were positive. Dengue virus genome RNA in female Aedes aegypti captured from dengue fever endemic areas in Hainan Island was detected by using RT-PCR, 1(33.33%) of 3 lots was positive. Similar examinations on bat brains and mosquitoes captured from non-endemic areas were all negative. In addition, monoclonal IgG antibodies against types 1-4 dengue viruses were added onto brain imprints of bats captured from endemic areas of dengue fever in Hainan Island, 16(80.00%) of 20 showed positive results for type 2 dengue virus antigen by direct immunofluorescent assay. The antibodies to various types of dengue virus were coincided to that of dengue virus genome RNA. The above results proved that bat is the reservoir of dengue virus, and this provides an important clue for the effective control of dengue fever epidemics in endemic areas.

Aedes↗

Fatal dengue hemorrhagic fever in adults during a dengue epidemic in Singapore.

BACKGROUND: Dengue fever has seen a significant re-emergence in Southeast Asia. Associated with the rise of dengue has been the increase in dengue-associated mortality. To better understand the predictors of mortality, we conducted a review of hospitalized adult dengue infections within our institution. METHODS: This was a retrospective case-control study of dengue-associated deaths at a large tertiary care hospital. RESULTS: In 2004, of 3186 cases of dengue fever (DF)/hemorrhagic dengue fever (DHF) admitted to our institution, there were 130 cases of DHF and seven dengue-associated deaths (case-fatality rate 5.4%). At least three of the seven fatal cases had serological evidence of primary dengue infection. All dengue-mortality cases had rapidly progressive clinical deterioration at an average of day 4 of fever with intensive care admission occurring on a mean of 5.6 days of fever. Adult respiratory distress syndrome, disseminated intravascular coagulopathy, and multi-organ failure were the most common causes of death despite early hospitalization, intravenous fluid, and blood-product support. CONCLUSION: Dengue is associated with severe disease, and deaths do occur despite current supportive management. Fatal DHF/dengue shock syndrome (DSS) does occur in adults and in primary dengue infection. Better early predictors of disease severity and clinical interventions are needed.

Adult↗

The 1986 dengue and dengue hemorrhagic fever epidemic in Puerto Rico: epidemiologic and clinical observations.

In 1986 Puerto Rico experienced its eleventh dengue outbreak of this century, but the first with simultaneous transmission of three dengue virus serotypes, and the first with significant numbers of severe and fatal hemorrhagic disease. Overall, 10,659 cases were reported; 1,257 cases were laboratory confirmed as having current or recent dengue infection. Dengue 4 (DEN-4) was the predominant serotype (160/363 isolates, 44%) followed by dengue 1 (DEN-1) with 134 isolates (37%) and dengue 2 (DEN-2), 69 isolates (19%). Transmission peaked during September, but large numbers of cases occurred through November. Seventy-one (91%) of Puerto Rico's 78 municipalities had laboratory-confirmed cases. Fifty-one percent of all confirmed cases occurred in metropolitan San Juan. Most cases presented clinically as classical dengue fever, but 37% of all confirmed cases were reported to have developed some type of hemorrhagic manifestation, and 6% reported hematemesis. In addition, 29 laboratory confirmed cases met the WHO case definition for dengue hemorrhagic fever, 3 of which were fatal. Among the 29 laboratory-confirmed cases of dengue hemorrhagic fever/ dengue shook syndrome, virus was isolated from 12; one DEN-1, three DEN-2, and eight DEN-4. Among laboratory confirmed cases, infants less than one year of age were at greater risk of developing dengue hemorrhagic fever/ dengue shook syndrome, hematemesis and any reported hemorrhage than were the other age groups evaluated.

Adolescent↗

Global situation of dengue and dengue haemorrhagic fever, and its emergence in the Americas.

About two-thirds of the world's population live in areas infested with dengue vectors, mainly Aedes aegypti. All four dengue viruses are circulating, sometimes simultaneously, in most of these areas. It is estimated that up to 80 million persons become infected annually although marked underreporting results in the notification of much smaller figures. Currently dengue is endemic in all continents except Europe and epidemic dengue haemorrhagic fever (DHF) occurs in Asia, the Americas and some Pacific islands. The incidence of DHF is much greater in the Asian countries than in other regions. In Asian countries the disease continues to affect children predominantly although a marked increase in the number of DHF cases in people over 15 years old has been observed in the Philippines and Malaysia during recent years. In the 1990's DHF has continued to show a higher incidence in South-East Asia, particularly in Viet Nam and Thailand which together account for more than two-thirds of the DHF cases reported in Asia. However, an increase in the number of reported cases has been noted in the Philippines, Lao People's Democratic Republic, Cambodia, Myanmar, Malaysia, India, Singapore and Sri Lanka during the period 1991-1995 as compared to the preceding 5-year period. In the Americas, the emergence of epidemic DHF occurred in 1981 almost 30 years after its appearance in Asia, and its incidence is showing a marked upward trend. In 1981 Cuba reported the first major outbreak of DHF in the Americas, during which a total of 344,203 cases of dengue were notified, including 10,312 severe cases and 158 deaths. The DHF Cuban epidemic was associated with a strain of dengue-2 virus and it occurred four years after dengue-1 had been introduced in the island causing epidemics of dengue fever. Prior to this event suspected cases of DHF or fatal dengue cases had been reported by five countries but only a few of them fulfilled the WHO criteria for diagnosis of DHF. The outbreak in Cuba is the most important event in the history of dengue in the Americas. Subsequently to it, in every year except 1983, confirmed or suspected cases of DHF have been reported in the Region. The second major outbreak in the Americas occurred in Venezuela in 1989 and since then this country has suffered epidemics of DHF every year. Between 1981 and 1996 a total of 42,246 cases of DHF and 582 deaths were reported by 25 countries in the Americas, 53% of which originated from Venezuela and 24% from Cuba. Colombia, Nicaragua and Mexico have each reported over 1,000 cases during the period 1992-1996. About 74% of the Colombian cases and 97% of the Mexican cases were reported during 1995-1996. A main cause of the emergence of DHF in the Americas was the failure of the hemispheric campaign to eradicate Aedes aegypti. Following a successful period that resulted in the elimination of the mosquito from 18 countries by 1962, the programme began to decline and as a result there was a progressive dissemination of the vector so that by 1997 with the exception of Canada, Chile and Bermuda, all countries in the Americas are infested. Other factors contributing to the emergence/re-emergence of dengue/DHF include the rapid growth and urbanization of populations in Latin America and the Caribbean, and increased travel of persons which facilitates dissemination of dengue viruses. Presently, all four dengue serotypes are circulating in the Americas, thus increasing the risk for DHF in this region.

Americas↗

Role of T cells, cytokines and antibody in dengue fever and dengue haemorrhagic fever.

Dengue infections are a major cause of morbidity and mortality in the tropical and sub-tropical regions of the world. There is no vaccine for dengue and also there are no anti-viral drugs to treat the infection. Some patients, typically those experiencing a secondary infection with a different dengue serotype, may progress from an acute febrile disease to the more severe forms of disease, dengue haemorrhagic fever and dengue shock syndrome. Here we discuss the significant immunopathological component to severe disease and how T cells, cytokines and cross-reactive antibody combine to contribute to the progression to dengue haemorrhagic fever. These events are thought to lead to vascular leakage, the signature event in dengue haemorrhagic fever, and are addressed in this review by incorporating the concept of heterologous T cell immunity. The need for effective measures against dengue and dengue-related illness is clear. We propose that drugs against dengue virus, or the symptoms of severe dengue disease, are a viable goal.

Animals↗

Failure of secondary infection with American genotype dengue 2 to cause dengue haemorrhagic fever.

BACKGROUND: Population-based epidemiological studies have shown that infection with dengue type 2 (DEN-2) virus in individuals previously infected with a different serotype of the virus is a major risk factor for dengue haemorrhagic fever and dengue shock syndrome. However, the western hemisphere was spared epidemics of these two syndromes, until the introduction of a southeast Asian DEN-2 genotype. Possibly American DEN-2 genotype strains lacked properties necessary to cause severe disease. We report on a major epidemic of DEN-2 in Peru in 1995, about 5 years after an epidemic of DEN-1 in the same population. METHODS: In Iquitos, a city of 344,686 inhabitants in Peru, cases of dengue fever were studied prospectively from 1990. Acute phase of illness serum samples from patients were tested for virus in C6/36 cells, and virus isolates were identified by immunofluorescence. Isolates of dengue 2 virus obtained from patients during an outbreak of mild febrile illness in 1995 were sequenced to determine the genotype. Serological analysis of paired samples from the patients was done with an IgM capture ELISA and an indirect IgG ELISA. In addition, serum samples collected annually between 1993 and 1996 from a large cohort of students were tested for dengue IgG antibody by an ELISA. Serum samples from a random sample of 129 students from this cohort were tested for dengue neutralising antibodies to quantify the serotype specific infection rates. FINDINGS: Among the 129 students (aged 7-20 years in 1993) who had serum samples available before and after the epidemic, 78 (60.5%) had a secondary DEN-2 virus infection. By extrapolation, 49,266 of the 81,479 children (aged 5-14 years) in Iquitos would have experienced such infections. From previous studies, between 887 and 10,247 cases of dengue haemorrhagic fever and dengue shock syndrome would have been expected. No cases were found. DEN-2 isolates were of the American genotype. INTERPRETATION: This prospective study shows that secondary infection by the American DEN-2 genotype did not cause dengue haemorrhagic fever and dengue shock syndrome.

Adolescent↗

Classifying dengue: a review of the difficulties in using the WHO case classification for dengue haemorrhagic fever.

BACKGROUND: The current World Health Organisation (WHO) classification of dengue includes two distinct entities: dengue fever (DF) and dengue haemorrhagic fever (DHF)/dengue shock syndrome; it is largely based on pediatric cases in Southeast Asia. Dengue has extended to different tropical areas and older age groups. Variations from the original description of dengue manifestations are being reported. OBJECTIVES: To analyse the experience of clinicians in using the dengue case classification and identify challenges in applying the criteria in routine clinical practice. METHOD: Systematic literature review of post-1975 English-language publications on dengue classification. RESULTS: Thirty-seven papers were reviewed. Several studies had strictly applied all four WHO criteria in DHF cases; however, most clinicians reported difficulties in meeting all four criteria and used a modified classification. The positive tourniquet test representing the minimum requirement of a haemorrhagic manifestation did not distinguish between DHF and DF. In cases of DHF thrombocytopenia was observed in 8.6-96%, plasma leakage in 6-95% and haemorrhagic manifestations in 22-93%. The low sensitivity of classifying DHF could be due to failure to repeat the tests or physical examinations at the appropriate time, early intravenous fluid therapy, and lack of adequate resources in an epidemic situation and perhaps a considerable overlap of clinical manifestations in the different dengue entities. CONCLUSION: A prospective multi-centre study across dengue endemic regions, age groups and the health care system is required which describes the clinical presentation of dengue including simple laboratory parameters in order to review and if necessary modify the current dengue classification.

Capillary Permeability↗