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Low rates of antigen detection and virus isolation from the peripheral blood leukocytes of dengue fever patients.

We evaluated direct fluorescent antibody (FA) testing of peripheral blood leukocytes (PBL) from patients in Puerto Rico with serologically and/or virologically confirmed dengue fever as a possible rapid diagnostic test and compared rates of dengue virus isolation from PBL with the rates from plasma or serum using the mosquito inoculation technique. Dengue antigen was detected in the PBL of only 1 of 19 patients with confirmed dengue. Virus was isolated from 3 of 19 PBL specimens and from 6 of 19 acute-phase serum or plasma samples. Four viruses were obtained from serum or plasma only and 1 isolate came from PBL only. We conclude that FA testing of PBL from dengue fever patients has little promise as a rapid diagnostic technique. Despite small numbers, our data suggest that virus isolation from PBL is less sensitive than that from serum or plasma. Our results differ considerably from those of previous studies of dengue hemorrhagic fever patients conducted in Thailand.

Antibodies, Viral↗

Dengue in Puerto Rico, 1977: public health response to characterize and control an epidemic of multiple serotypes.

The largest and most extensive documented dengue epidemic in Puerto Rico struck an estimated 355,000 Puerto Rican residents from July-December 1977. The mixed epidemic of dengue types 2 and 3 coincided with a Caribbean pandemic of dengue type 1, first introduced into the western hemisphere in early 1977 and into Puerto Rico in the fall of that year. Health officials assembled a team to assess the epidemic and mounted a campaign to end it. Attempts to monitor the incidence and spread of dengue were confounded by simultaneous co-circulation of influenza virus, underscoring problems in formulating public health strategies dependent on nonspecific clinical and epidemiologic case criteria, and the need for rapid and reliable diagnostic capabilities. Despite co-circulation of multiple dengue serotypes, a risk factor associated with severe and fatal dengue hemorrhagic fever (DHF) in Southeast Asia, hospital and death certificate surveillance disclosed no cases of DHF in Puerto Rico. The epidemic serves as a reminder that when preventive measures are impossible or infeasible, developed countries with high living standards may be susceptible to large scale epidemics of infectious diseases.

Aedes↗

Epidemic of dengue-4 virus in Yap State, Federated States of Micronesia, and implication of Aedes hensilli as an epidemic vector.

A dengue fever/dengue hemorrhagic fever (DF/DHF) outbreak in Yap State caused by dengue-4 virus was confirmed serologically and by virus isolation from serum samples collected on each of three island groups. Most DF/DHF cases occurred during a three-month period between mid-May and early August 1995. Five fatal cases, three of which were in children between the ages of four and 11, occurred between June 20 and July 26. A serosurvey conducted in late August revealed anti-dengue IgM prevalence rates of 18% on Yap, 36% on Eauripik, and 6% on Woleai. The majority of residents (93-100%) on the three islands were positive for anti-dengue IgG antibodies, indicating widespread exposure to dengue viruses. The IgG titers indicative of secondary antibody response were noted on Eauripik (6.5%) and Woleai (17%), but were rare on Yap (0.7%). Entomologic investigations implicated the native mosquito species, Aedes hensilli, a member of the Scutellaris Group of Aedes (Stegomyia), as a previously unrecognized epidemic vector of dengue viruses. Aedes hensilli was the most abundant and widespread member of Ae. (Stegomyia) in Yap State, the only species of Ae. (Stegomyia) on Woleai, and the only mosquito species present on Eauripik. New distribution records for mosquito species are reported.

Adolescent↗

Detection and genetic relationship of dengue virus sequences in seventeen-year-old paraffin-embedded samples from Cuba.

This study describes the use of the reverse transcriptase-polymerase chain reaction (RT-PCR) to generate dengue 2 amplicons from paraffin-embedded autopsy tissues collected in Cuba 17 years ago. The presumptive diagnoses had been made only by clinical evolution without serologic confirmation. This study confirms once again that dengue 2 virus was directly associated with the fatal cases in children and illustrates the potential of the RT-PCR for retrospective diagnosis of dengue cases 17 years after death. A close similarity in the genomic sequences of the dengue 2 RNA detected in tissue samples from fatal cases and those dengue 2 Cuban strains that had been previously investigated confirms the appropriate genomic classification of the etiologic agent associated with the 1981 dengue hemorrhagic fever Cuban epidemic.

Amino Acid Sequence↗

Clinical, epidemiologic, and virologic features of dengue in the 1998 epidemic in Nicaragua.

From July to December 1998, a hospital- and health center-based surveillance system for dengue was established at selected sites in Nicaragua to better define the epidemiology of this disease. Demographic and clinical information as well as clinical laboratory results were obtained, and virus isolation, reverse transcriptase-polymerase chain reaction, and serologic assays were performed. World Health Organization criteria were used to classify disease severity; however, a number of patients presented with signs of shock in the absence of thrombocytopenia or hemoconcentration. Therefore, a new category was designated as "dengue with signs associated with shock" (DSAS). Of 1,027 patients enrolled in the study, 614 (60%) were laboratory-confirmed as positive cases; of these, 268 (44%) were classified as dengue fever (DF); 267 (43%) as DF with hemorrhagic manifestations (DFHem); 40 (7%) as dengue hemorrhagic fever (DHF); 20 (3%) as dengue shock syndrome (DSS); and 17 (3%) as DSAS. Interestingly, secondary infection was not significantly correlated with DHF/DSS, in contrast to previous studies in Southeast Asia. DEN-3 was responsible for the majority of cases, with a minority due to DEN-2; both serotypes contributed to severe disease. As evidenced by the analysis of this epidemic, the epidemiology of dengue can differ according to geographic region and viral serotype.

Adolescent↗

Control of aedes vectors of dengue in three provinces of Vietnam by use of Mesocyclops (Copepoda) and community-based methods validated by entomologic, clinical, and serological surveillance.

We describe remarkable success in controlling dengue vectors, Aedes aegypti (L.) and Aedes albopictus (Skuse), in 6 communes with 11,675 households and 49,647 people in the northern provinces of Haiphong, Hung Yen, and Nam Dinh in Vietnam. The communes were selected for high-frequency use of large outdoor concrete tanks and wells. These were found to be the source of 49.6-98.4% of Ae. aegypti larvae, which were amenable to treatment with local Mesocyclops, mainly M. woutersi Van der Velde, M. aspericornis (Daday) and M. thermocyclopoides Harada. Knowledge, attitude, and practice surveys were performed to determine whether the communities viewed dengue and dengue hemorrhagic fever as a serious health threat; to determine their knowledge of the etiology, attitudes, and practices regarding control methods including Mesocyclops; and to determine their receptivity to various information methods. On the basis of the knowledge, attitude, and practice data, the community-based dengue control program comprised a system of local leaders, health volunteer teachers, and schoolchildren, supported by health professionals. Recycling of discards for economic gain was enhanced, where appropriate, and this, plus 37 clean-up campaigns, removed small containers unsuitable for Mesocyclops treatment. A previously successful eradication at Phan Boi village (Hung Yen province) was extended to 7 other villages forming Di Su commune (1,750 households) in the current study. Complete control was also achieved in Nghia Hiep (Hung Yen province) and in Xuan Phong (Nam Dinh province); control efficacy was > or = 99.7% in the other 3 communes (Lac Vien in Haiphong, Nghia Dong, and Xuan Kien in Nam Dinh). Although tanks and wells were the key container types of Ae. aegypti productivity, discarded materials were the source of 51% of the standing crop of Ae. albopictus. Aedes albopictus larvae were eliminated from the 3 Nam Dinh communes, and 86-98% control was achieved in the other 3 communes. Variable dengue attack rates made the clinical and serological comparison of control and untreated communes problematic, but these data indicate that clinical surveillance by itself is inadequate to monitor dengue transmission.

Adolescent↗

Differentiating dengue virus infection from scrub typhus in Thai adults with fever.

Dengue fever and scrub typhus are common infections in Asia that often present as acute febrile illness of unclear etiology. We prospectively evaluated febrile adults presenting to the outpatient department of a hospital in northern Thailand to attempt to identify distinguishing characteristics between the two infections. Fifty-four patients were infected with scrub typhus and 35 were infected with dengue virus. Dengue virus infection was associated with hemorrhagic manifestations, particularly bleeding from the gums, which was reported by 27% of the dengue patients, but by none of the scrub typhus patients (P < 0.001, by Fisher's exact test). A low platelet count (< 140,000/mm3) and low white blood cell count (< 5,000/mm3) were strongly associated with dengue infections: odds ratio = 26.3 (95% confidence interval [CI] = 7.4-93.2) for platelet count and 8.2 (95% CI = 2.6-25.5) for leukocyte count. Prospective evaluations of the usefulness of these simple criteria to differentiate scrub typhus from dengue infection are needed in other areas, particularly where rapid confirmatory diagnostic tests are not available.

Adult↗

Induction of programmed cell death (apoptosis) by dengue virus in vitro and in vivo.

Dengue is a human disease which may be fatal in its hemorrhagic form. How dengue virus- and host-specified factors underlie virulence and pathogenesis is poorly understood. An immunological disorder is thought to be involved in dengue physiological symptoms. Whether the immune response is deleterious or beneficial to the host remains a matter of debate. In this review, we summarized developments in research on viral pathogenesis in the context of apoptosis triggered by dengue virus infection. Apoptosis, an active process of cell destruction, is one of the important consequences of dengue virus infection in vitro and in vivo. Dengue virus replication induces apoptosis in mouse neurons and human hepatocytes. The ability to activate this genetically programmed cell death pathway is dependent on both viral and cellular determinants.

Animals↗

Comments on the epidemiology, pathogenesis and control of dengue.

Dengue is currently the most important viral disease transmitted to man by arthropods, whether measured by the number of cases or number of deaths. Prevalence of the disease is highest in tropical Asia, intermediate in tropical America, and lowest in tropical Africa. Four distinct dengue viruses have been identified. Types 2 and 3 appear to be more pathogenic on the average than types 1 and 4, but all four can cause severe or fatal dengue syndromes. Infection with any of the four viruses confers life-long homotypic, but not heterotypic, immunity. Dengue viruses can be transmitted by several mosquito species of the genus Aedes, but by far the most common vector is Aedes aegypti. All non-human primates that have been tested are susceptible to infection, but none exhibit signs of illness. The resulting lack of a suitable experimental host other than man has slowed progress in understanding the pathogenesis of severe forms of the disease. Controversy continues over the hypothesis that a first dengue infection increases the risk of severe disease upon re-infection as well as over the principal sites of replication. With regard to replication sites, some authors have mentioned cells of the mononuclear phagocyte lineage while others have implicated hepatocytes. No vaccine is currently available for dengue. Because the principal mosquito vector can use a wide variety of small domestic containers, vector control programs have not been highly effective.

Aedes↗

[Dengue fever].

Dengue virus infections have increased globally. This results in an increased incidence of patients with dengue fever in Denmark. In order to draw attention to this diagnosis among febrile persons returning from a stay in tropical and subtropical areas, we present two cases of dengue virus infection and describe the symptoms and diagnostic possibilities. Dengue Haemorrhagic Fever (DHF) is supposed to occur only in persons who have previously had dengue virus infection and who acquire a second infection with dengue virus.

Adult↗

[Dengue virus-specific antigen lymphoproliferation assay with human T cells].

Papers dealing with the study of the immune response to dengue virus infection and with the role it may play in the pathogenesis of dengue hemorrhaghic fever are increasingly important. The memory human T cell response in individuals with history of dengue infection during the Cuban epidemic was studied in this paper for a further evaluation of the antigenicity of viral proteins. To this end, mononuclear cells of peripheral blood from individuals immune to dengue and from a group of control subjects with viral antigens were incubated. It was obtained a significant proliferative response of lymphocytes from individuals with history of infection against dengue virus type 2 compared with control subjects. It was proved this way the dengue virus-specific memory T CD4+ cell response in the individuals under study.

Adult↗

Predominance of the DEN-3 genotype during the recent dengue outbreak in Bangladesh.

A recent outbreak of dengue in Bangladesh was marked by many fatal complications. As clinical virulence varies among the genotypes of dengue virus, a study was conducted to investigate the molecular genotypes of dengue in Bangladesh. Reverse transcription polymerase chain reaction was used to determine viral genotypes using oligonucleotide generic primers that produce a 511 bp product. The resulting product was typed by nested PCR with strain-specific primers, yielding 482 (DEN-1), 119 (DEN-2), 290 (DEN-3) and 392 (DEN-4), visualized on UV transilluminator after electrophoresis on 2% agarose gel stained with ethidium bromide. Of 45 clinically diagnosed dengue patients (mean age 28 years; male/female 30/15), 19 (42.2%) had detectable viral RNA in their blood. However, during the first 5 days of fever in 30 patients, the frequency was 60% (18/30), implying that the sooner serum is drawn after the fever, the greater the chances of detecting viral RNA. DEN-3 was detected in all except 2 patients who were infected with DEN-2. DEN-2 (two cases) and DEN-4 (one case) were present as co-infections with DEN-3. All of the patients presented with fever, anorexia and vomiting; many had headache and general body ache; a few had a rash. About a quarter had suffered episodes of bleeding, while ascites, pleural effusion and CNS symptoms were found in a few patients Patients positive for viral RNA were also positive for anti-dengue IgM (p=0.007) in subsequent sampling. The study suggests the predominance of DEN-3 infection with occasional co-infection with other types, during the recent outbreak of dengue in Bangladesh.

Adolescent↗

Serological & virological investigation of an outbreak of dengue fever in Gwalior, India.

BACKGROUND & OBJECTIVES: An outbreak of febrile illness occurred between September to November 2001 in Gwalior, Madhya Pradesh affecting individuals mostly in the age group < 30 yr. A total of 312 febrile indoor patients suspected to have dengue infection were investigated. METHODS: The investigation included examination of blood samples from patients for dengue specific IgM and IgG antibodies, isolation of virus in suckling mouse pups and in C(6/36) cell line followed by confirmation and typing through reverse transcriptase-PCR and nested PCR. RESULTS: The serological analysis of the 312 samples indicated 65 per cent positivity of which 21 per cent are of recent infection as indicated by the presence of IgM antibody and 78 per cent are found to be secondary in nature by showing the presence of IgG and/or IgM antibodies. The RT-PCR analysis of patients' sera employing dengue virus group specific conserved amplimer confirmed the etiological agent as dengue complex by showing the characteristic 511 bp amplicons. None of the antibody positive samples were found to be positive by RT-PCR. A total of 13 (6%) samples positive by RT-PCR, were processed for virus isolation in mouse pups and in C(6/36) cells. Of these 9 samples (80%) were confirmed positive for virus isolation as identified by RT-PCR. INTERPRETATION & CONCLUSION: The typing of isolates by nested PCR employing serotype specific amplimer revealed 119 bp amplicon characteristic of dengue virus type-2 and thus confirming the outbreak attributed to dengue virus type-2.

Adult↗

Variable susceptibility of the owl monkey (Aotus nancymae) to four serotypes of dengue virus.

The goal of this study was to determine, for each of four dengue serotypes, whether owl monkeys (Aotus nancymae) become viremic and develop antibody responses in patterns similar to those seen in humans and whether any behavioral parameters are reliably associated with immunologic responses. A secondary goal was to investigate effects of chronic blood sampling on hematologic parameters in this genus. We inoculated 20 owl monkeys with 2 x 10(4) plaque-forming units of one of four dengue serotypes. Blood samples ranging from 0.4 to 0.7 ml per animal were taken each day for 12 consecutive days after inoculation, as well as on days 21, 28, and 60 post-inoculation. The total amount of blood taken per monkey was 8.0 ml during the first 12 days and 9.5 ml during the first 30 days of the study (i.e., up to 17% total blood volume per week and up to 20% total blood volume per month). Detailed behavioral assessments of all animals were made twice daily on every day of sample collection. The dengue-1 group were viremic for an average of 3.75 days. Dengue-2, -3, and -4 groups had average viremias of 1.00, 1.25, and 1.33 days, respectively. All animals demonstrated appropriate antibody responses as determined by enzyme-linked immunosorbency assay (ELISA). Animals tolerated repeated phlebotomy well, as all animals remained within clinically normal hematocrit (HCT) reference ranges, and no lasting effects on HCT occurred in any monkey. Final HCT for most animals was greater than 45% (mean final hematocrit, 45%). The maximum decrease in HCT ranged from 3.5 to 19 (mean, 8.9) percentage points. No consistent correlation of any behavioral disease parameters with viremia and antibody status was demonstrated, although overt illness did occur in two animals. Aotus can be an affordable and safe model for testing dengue vaccine efficacy; further testing with higher doses of dengue-2, -3 and -4 viruses is warranted.

Animals↗

Studies on dengue in rural areas of Kurnool District, Andhra Pradesh, India.

A dengue case was reported for the 1st time in a rural area of Kurnool District, Andhra Pradesh, India. Entomological and serological investigations were carried out to determine the prevalence of dengue vectors and dengue virus. Aedes aegypti was recorded for the 1st time in rural areas of Andhra Pradesh. Breeding of Ae. aegypti was observed only in containers with nonpotable water. Cement cisterns and tanks, stone tubs, and clay pots were the major breeding habitats of Ae. aegypti. Larval indices for Ae. aegypti ranged as follows: house index 28-40%, container index 13-37%, and Breteau index 32-60. A serological survey indicated that humans in Kurnool District have been exposed to dengue virus infections. The potential threat of an outbreak of dengue fever in rural areas because of the prevalence of the vector (Ae. aegypti) and dengue virus is discussed.

Aedes↗

Perinatal dengue infection: a case report and review of literature.

A case of vertical transmission of dengue infection in the perinatal period is reported. The mother, a term pregnancy, had acute dengue the day before admission. The infant was born at term and developed fever on the fifth day of life which lasted for 5 days. No bleeding or plasma leakage was detected during the course of fever in infant or mother. A liver function test showed elevated SGOT and SGPT in the infant. The infant developed a convalescent rash on day 5 of the fever. The diagnosis of secondary dengue hemorrhagic fever in the mother was confirmed by serology and primary dengue infection in the infant was confirmed by serology and serotyped as dengue type 2 by PCR. The clinical course and management of mothers and infants with perinatal dengue infection are reviewed.

Acute Disease↗

Diagnosis of dengue infection using various diagnostic tests in the early stage of illness.

In order to elucidate the usefulness of various tests in the early course of dengue infection, in terms of diagnosis and correlation with clinical severity, blood specimens were collected every 48 hours on 3 occasions from patients with clinical suspicion of dengue infection with fever for less than 4 days. Viral isolation was attempted by mosquito inoculation (MI), tissue culture inoculation (TC), and reverse transcriptase polymerase chain reaction (RT-PCR). Antibodies were detected by hemagglutination inhibition test (HI), an in-house-ELISA (IH-ELISA), and an ELISA by MRL diagnostics Clinical data were collected from the time of enrollment to complete recovery. Of the 40 patients enrolled, 31 were diagnosed as dengue infection and confirmed by either serology or viral isolation. Of these, 12 had primary infection and 19 had secondary infection. Dengue fever occurred in 9 cases. Dengue viruses were isolated from 28 out of 31 patients, and dengue hemorrhagic fever was diagnosed in 22 patients. Viral serotypes identified by viral isolation, and RT-PCR were concordant: DEN1 was isolated in 8, DEN2 in 13, DEN3 in 5, and DEN4 in 2 patients. Viral isolation yielded positive results on blood collected before the 5th day of fever. MI was more sensitive than TC. RT-PCR was less sensitive than viral isolation during the early days of fever, but became more sensitive after the 5th day of fever. RT-PCR was able to detect virus up to day 7-8 of fever, even after defervescence, and in the presence of antibody. During the febrile stage, serological diagnosis on blood samples taken 48 hours apart was carried out by HI, IH-ELISA, and MRL-ELISA, facilitating diagnosis in 3 (10%), 21 (67%), and 27 (87%) of patients, respectively. All of the patients with secondary infection were diagnosed by MRL-ELISA before defervescence. By the 8th day of fever, a serological diagnosis aided to diagnose in 9 (29%), 29 (93%), and 31 (100%) of patients by HI, IH-ELISA, and MRL-ELISA, respectively.

Adolescent↗

[Impact on the potential epidemic of dengue fever under warming winter in Hainan province].

OBJECTIVE: Impact of climate warming in winter on the potential epidemics of dengue fever in Hainan was assessed. METHODS: Based on historic data of mean monthly temperature in January from 8 weather observation stations, tendency and amplitude of variation were analyzed. Using 21 degrees C as lowest limit of temperature suitable for dengue fever transmission, impact caused by climate warming on dengue fever epidemic was estimated by means of geography information system (GIS), insect vector and epidemiological features. RESULTS: Temperature in winter in Hainan province had shown an obvious increase. The maximum amplitude of increase appeared in Dongfang which was 1.4 degrees C and the minimum 0.5 degrees C in Shanhudao, but the increase amplitude in the other stations was varied from 0.7 to 1.3 degrees C. By the year of 2050, 21 degrees C contour will have moved 190 km or so northward, nearly spanned 6/7 of distance from south to north in Hainan province and under the condition of daily fraction surviving of Aedes aegypti as P = 0.89, Qionghai city which stands north in Hainan province will probably have become epidemic area of dengue fever all year round. CONCLUSION: Climate warming in winter will probably make half or more of the areas in Hainan province with temperature that permitting transmission of dengue fever by 2050. Monitoring and prevention of dengue fever in winter should be emphasized.

Aedes↗